Antioxidative enzymes in resistant and susceptible genotypes of tomato infected with root-knot nematode

Vol. 43, No. 1, pp. 1-12 Indian Journal of Nematology June, 2013

Changes in Antioxidative Enzymes in Resistant and Susceptible Genotypes of

Tomato Infected with Root-Knot Nematode (Meloidogyne incognita)
NEENA CHAWLA, KAVITA CHOUDHARY**, SUKHJEET KAUR* AND SALESH JINDAL

Department of Vegetable Crops, **Department of Biochemistry, Punjab Agricultural University, Ludhiana-141004, India
*Corresponding author, E-mail: randhawask@[Link]

Received on 17-05-2012 and Accepted on 06-01-2013

ABSTRACT: Changes in activities of antioxidative enzymes (Catalase, Peroxidase, Superoxide dismutase, Polyphenol oxidase,
esterase, monodehydro ascorbate reductase, and dehydro ascorbate reductase) after nematode inoculation were investigated in four
varieties of tomatoes, (two resistant i.e. Hisar Lalit, PNR- 7 and two susceptible i.e. Punjab Varkha Bahar-1 and Punjab Varkha Bahar-
2. These four varieties were infected at the nursery stage with root-knot nematode (Meloidogyne incognita). The egg masses of M.
incognita were taken from the infected brinjal plants. Tomato roots and leaves from un-inoculated and inoculated plants were
evaluated for their antioxidative enzyme activities after 7 days and 20 days of infection. The activity of antioxidant enzymes increased
after infection with root-knot nematode in the resistant varieties of tomato as compared to that of susceptible varieties. The activities
of all enzymes increased after inoculation except the activity of CAT enzyme where a decrease was observed. The isozyme banding
patterns of peroxidase and esterase showed that the intensity of bands was more in the resistant genotypes as compared to that of
susceptible genotypes, which means that the activity of antioxidant enzymes were more after inoculation in the resistant genotypes
as compared to that of susceptible genotypes. An additional band was obtained in the isozyme banding pattern of esterase in the
resistant genotypes.

Keywords: Tomato, nematode, Meloidogyne incognita, antioxidant enzymes, isozyme

Tomato (Lycopersicon esculentum Mill.) a member usually associated with hypersensitive reaction (HR), a
of family Solanaceae, is universally known as a protective rapid and localized cell death in the infected plant in
food. It is used directly as raw vegetable, in sandwiches, response to nematode attack.
salads etc. and also as purees, chutneys and pickles in
processed forms. Tomato fruit has undoubtedly assumed Reactive oxygen species (ROS) play an important
the status of a functional food considering the role in plant defense and during pathogen attack, levels
overwhelming epidemiological evidence for its anti- of ROS detoxifying enzymes like peroxidase (POX) and
cancer activity. Root-knot nematode (Meloidogyne catalase (CAT) are often suppressed in resistant plants
incognita) causes considerable damage to tomato plants (Klessing et al., 2000). As a result, plants produce more
(Kamalvanshi et al., 2004) and results in extensive ROS and accumulation of these components leads to HR
losses in yield and quality (Mahajan & Singh, 2001). in plant cells. For example, hydrogen peroxide plays a
Though the nematicides are effective against nematodes, major role in triggering HR in incompatible interactions
yet they are hazardous for health, soil and environment. (Dangl & Jones 2001). Antioxidant enzymes such as
Plant resistance is one of the eco-friendly options for the SOD, POD and CAT are considered to be the main
management of nematode diseases. A series of protective enzymes engaged in the removal of free
biochemical and physical reactions occur in plants in radicals and activated oxygen species (Blokhina et al.,
response to root-knot nematode infection. Plants 2003, Devi et al., 2000). Oxidative enzymes such as
synthesize certain compounds that are toxic to root-knot POX and pro are reported to be involved in the mechanism
nematode. In the family of solanaceae, resistance is of disease resistance.

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Neena Chawla et al.

Peroxidases are heme containing enzymes that growing in 30 cm diameter earthen pots for further
catalyse one electron oxidation of several substrates at multiplication. One week old seedlings of tomato genotypes
the expense of hydrogen peroxide and are located in cell sown in pots were inoculated with freshly hatched larvae
walls and vacuoles. The role of peroxidase in producing of root knot nematode (M. incognita) @ 1 J2s/cc of soil.
active oxygen species (AO) is controversial. In cell wall Leaf and root samples were collected at 7 and 20 days
peroxidase oxidizes NAD(P)H to produce O2- and H2O2 after inoculation with root-knot nematode.
(Elstner and Heupel 1976). Peroxidase in its activity uses Simultaneously, root and leaf samples were also collected
H2O2 as the main oxidant and reduces it into H2O, thus from uninoculated plants and carried to the laboratory in
acting as an active AO scavenger. Electrophoretic an ice box and analyzed for anti oxidative enzymes.
analysis of peroxidase revealed that both the resistant
and susceptible plants responded to parasitic invasion by Sample was homogenized in 5ml of cold (40 C)
synthesizing new peroxidases isozymes. SOD, the family extraction buffer using pre chilled pestle and mortar. The
of metallo-enzymes, catalyses the disproportionation of mixture was then centrifuged at 20,000g for 10 minutes
superoxide O2- to molecular oxygen and H2O2 (Scandalios and supernatant was collected and was analyzed for
1993). enzyme activity.

Catalase catalyses breakdown of H2O2 using one Catalase- To chilled sodium phosphate buffer added
molecule of H2O2 as substrate donor and another molecule enzyme extract. The reaction was started by adding
of H2O2 as oxidant or electron acceptor. Since catalase H2O2 to the reaction mixture. The rate of decrease in
repression results in H2O2 accumulation, this molecule absorbance at 240 nm was measured at 10 second
may be responsible for the production of defense signals intervals for 1 min (Chance and Mahley 1955).
that directly trigger HR. Two enzymes are involved in
the regeneration of reduced ascorbate, namely mono- Superoxide Dismutase- The reaction mixture contained
dehydro-acsorbate reductase (MDHAR) which uses methionine, NBT , EDTA, Na2CO3, phosphate buffer
NAD(P)H directly to recycle ascorbate and dehydro- and distilled water. The enzyme extract was added at
ascorbate-reductase (DHAR). However, the situation is last and the reaction was started by adding 0.1ml riboflavin.
further complicated because mono-dehydro-ascorbate The intensity of the color developed was read at 560nm
itself is an efficient electron acceptor (Foyer and (Xing et al., 2008).
Lelandais 1993, Miyake and Asada 1992). Mono-
dehydro-ascorbate is reduced directly to ascorbate using Polyphenol Oxidase- To enzyme extract, phosphate
electrons derived from the photosynthetic electron buffer was added. Reaction was started by adding 2% of
transport chain. catechol. PPO activity was assayed by measuring the
linear increase in absorbance at 410nm (Augustin et al.,
MATERIALS AND METHODS 1985).

The seeds of tomato genotypes viz: (Hisar lal, PNR- Dehydroascorbate Reductase- The reaction mixture
7, Punjab Varkha Bahar-l and Punjab Varkha Bahar-2) contained 0.1M phosphate buffer, ImM EDTA, 2.5mM
were procured from the Department of Vegetables glutathione reduced, 0.2mM DHA. To this enzyme
Crops, Punjab Agricultural University, Ludhiana. extract was added. DHAR was assayed by measuring
the reduction of dehydro-ascorbate at 265nm with an
Egg masses of Meloidogyne incognita were extinction coefficient of 14 (Foyer et al., 1989).
collected from pure culture already maintained on brinjal
plants. These egg masses were transferred to tissue Esterase- To enzyme extract, phosphate buffer was
paper kept on wire gauge suspended in water in petri added. Reaction was started by adding 10-3 M indophenyl
plate, to allow the eggs to hatch. The second stage acetate. Esterase activity was assayed by change in
juveniles (J2s) were then inoculated to brinjal plants absorbance at 625 nm (Sae et al., 1971).

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 Antioxidative enzymes in resistant and susceptible genotypes of tomato infected with root-knot nematode

Table 1: Superoxide dismutase (SOD) activity (unit SOD g-l FW) in tomato roots (R) and leaves (L) of resistant and susceptible
genotypes against root-knot nematode

Genotype Days after inoculation

(Specific activity of SOD)
7 20
U I U I

Resistant genotypes
Hissar lalit (R) 0.301±0.007 0.311±0.012 0.316±0.013 0.623±0.015
(0.071±0.005) (0.076±0.006) (0.074±0.007) (0.134±0.008)
PNR-7 (R) 0.416±0.014 0.42800.016 0.527±0.015 0.831±0.016
(0.104±0.007) (0.113±0.006) (0.132±0.007) (0.205±0.005)
Hisar lalit (L) 0.361±0.014 0.372±0.013 0.431±0.017 0.798±0.018
(0.069±0.006) (0.072±0.006) (0.082±0.006) (0.148±0.008)
PNR-7 (L) 0.346±0.016 0.35100.015 0.398±0.016 0.842±0.014
(0.059±0.004) (0.061±0.005) (0.068±0.006) (0.134±0.008)
Susceptible genotypes
Varkha Bahar 1 (R) 0.211±0.016 0.223±0.010 0.275±0.016 0.301±0.007
(0.032±0.002) (0.034±0.002) (0.042±0.003) (0.051±0.004)
Varkha Bahar 2 (R) 0.239±0.013 0.251±0.012 0.306±0.014 0.377±0.014
(0.033±0.002) (0.03500.003) (0.043±0.003) (0.054±0.004)
Varkha Bahar 1 (L) 0.224±0.015 0.235±0.014 0.268±0.015 0.303±0.016
(0.028±0.001) (0.031±0.002) (0.034±0.002) (0.042±0.003)
Varkha Bahar 2 (L) 0.271±0.017 0.280±0.015 0.298±0.016 0.316±0.015
(0.031±0.002) (0.035±0.002) (0.045±0.003) (0.052±0.004)

R= Roots; L= Leaves; U= Uninoculated plants; I= Inoculated plants

CD (P<0.05) R L R L R L
A 0.792 0.907 C 0.792 0.907 ABC 0.224 0.256
B 0.112 0.128 AC 0.112 0.128
AB 0.158 0.181 BC 0.158 0.181
Values are mean ± S.E. of three determinations
Values in parenthesis indicate specific activity (μmoles per mg of the protein) of the enzyme

Peroxidase- To chilled guaiacol added enzyme extract. started by adding 0.4 units of ascorbate oxidase to the
The reaction was started by adding H2O2 and the rate of reaction mixture. MDHAR was assayed by decrease in
decrease in absorbance at 470 nm was measured at 30 absorbance monitored at 340 nm using 6.2 as extinction
seconds intervals for 3 mins. The unit of enzyme is coefficient (Hossain & Asada, 1984 ).
defined as a decrease in O.D by 1.0 under standard
conditions (Shannon et al., 1966). Electrophoresis (Native-Page)- Electrophoresis was
done by according to Schagger et al (1994) method.
Monohydroascorbate Reductase- The reaction Dissolve the sample extract in equal volume of 2x sample
mixture contained Hepes buffer (pH 7.6), 1mM NADPH, buffer. Carefully pour the freshly prepared separating
2.5mM ascorbate and 50μl extract. The reaction was gel solution into the chamber without generating bubbles.

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Neena Chawla et al.

μmoles of H2O2 decomposed min-1 g-1 FW) in tomato roots (R) and leaves (L) of resistant and
Table 2: Catalase activity (CAT) (μ
susceptible genotypes against root-knot nematode

Genotype Days after inoculation

(Specific activity of Catalase)
7 20
U I U I

Resistant genotypes
Hisar lalit (R) 98.75±2.11 84.63±3.12 75.21±4.14 68.39±5.09
(23.22±1.89) (20.84±1.37) (17.69±1.09) (15.06±1.19)
PNR-7 (R) 101.52±3.91 84.32±4.19 74.61±2.95 54.51±4.16
(25.57±1.19) (22.30±1.37) (18.79±1.34) (10.88±0.57)
Hisar lalit (L) 144.16±:4.97 109.84±2.84 117.56±:3.79 98.42±5.14
(27.72±1.84) (21.53±1.14) (22.60±1.18) (18.25±0.98)
PNR-7 (L) 151.77±:3.46 116.75±2.66 123.90±1.77 102.77±4.59
(25.89±1.65) (20.59±1.54) (21.14±1.67) (16.46±1.02)
Susceptible genotypes
Varkha Bahar l (R) 48.73±3.40 59.26±3.98 54.17±3.04 66.30±3:11
(7.47±0.21) (9.21±0.22) (830:±0.22) (10.01±0.23)
Varkha Bahar 2 (R) 64.97±2.31 78.17±4.19 73.90±1.41 89.20±2.96
(9.16±0.21) (11.16±0.24) (10.43±0.23) (11.79±0.25)
Varkha Bahar 1 (L) 103.55±2.58 121.85±2.47 142.89±2.75 153.25±4.10
(13.19±0.22) (15.72±0.23) (18.20±0.20) (19.47±0.21)
Varkha Bahar 2 (L) 126,69±2.06 134.51±3.63 166.30±2.83 177.74±2.91
(15.89±0.23) (20.44±1.69) (20.86±1.77) (24.82±1.84)

R= Roots; L= Leaves; U= Uninoculated plants; I= Inoculated plants

CD (P<0.05) R L R L R L
A NS 2.017 C 2.058 2.017 ABC NS 5.705
B 2.910 2.852 AC NS 2.852
AB 4.116 4.034 BC 4.116 4.034
Values are mean ± S.E. of three determinations
Values in parenthesis indicate specific activity (μmoles per mg of the protein) of the enzyme

Carefully overlayer the acrylamide solution with H2O- plates. Stain the gel in the staining solution for 2-3 hours.
saturated n-Butanol without mixing. Polymerize the gel Remove the dye that was not bound to protein in
for 1 h. Pour off the n-butanol from the polymerized destaining solution. The bands of the isozymes appeared
separating gel, wash the gel top with water and fill the in the gel. Finally the photographs were taken to have the
gap remaining in the chamber with stacking gel solution, permanent record of the isozymes banding patterns.
insert the comb. When the stacking gel has polymerized,
remove the comb without distorting the shapes of the RESULTS AND DISCUSSION
well. Fill the apparatus reservoir buffer. Load the gel
with 10-30 μl sample solution by pipette. Start the The SOD activity in the roots and leaves of both
electrphoresis immediately by turning on power. After inoculated as well as uninoculated plants of resistant and
electrophoresis, remove the gel from between the glass susceptible genotypes increased after 20 days of infection

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 Antioxidative enzymes in resistant and susceptible genotypes of tomato infected with root-knot nematode

Table 3: Peroxidase activity (POX) (units min-1 mg-1 FW) in tomato roots (R) and leaves (L) of resistant and susceptible genotypes
against root-knot nematode

Genotype Days after inoculation

(Specific activity of peroxidase)
7 20
U I U I

Resistant genotypes
Hisar lalit (R) 1.04±0.08 3.85±0.27 4.17±0.32 8.64±0.39
(0.244±0.021) (0.948±0.026) (0.981±0.022) (1.903±0.025)
PNR-7 (R) 2.06±0.11 5.10±0.42 4.28±0.39 9.37±0.64
(0.518±0.028) (1.349±0.029) (1.078±0.024) (1.870±0.31 )
Hisar lalit (L) 4.55±0.38 5.60±0.44 7.45±0.58 10.10±0.68
(0.875±0.019) (1.098±0.021) (1.432±0.023) (1.873±0.031)
PNR-7 (L) 5.09±0.44 7.15±0.61 7.07±0.68 8.84:±0.69
(0.868±0.032) (1.261±0.034) (1.206±0.029) (1.416±0.033)
Susceptible genotypes
Varkha Bahar 1 (R) 3.28±0.21 2.49±0.13 2.64±0.15 1.98±0.07
(0.503±0.031) (0.387±0.026) (0.404±0.035) (0.299±0.025)
Varkha Bahar 2 (R) 4.78±0.35 3.58±0.27 4.00±0.32 3.09±0.25
(0.674±0.033) (0.511±0.022) (0.564±0.036) (0.408±0.031 )
Varkha Bahar 1 (L) 5.31±0.47 4.52±0.30 4.37±0.29 3.95±0.23
(0.676±0.032) (0.583±:0.028) (0.556±0.037) (0.501±0.034)
Varkha Bahar 2 (L) 9.32±0.56 8.57±0.46 8.16±0.61 6.60±0.44
(1.416±0.026) (1.075±0.023) (1.240±0.035) (0.921±0.024)

R= Roots; L= Leaves; U= Uninoculated plants; I= Inoculated plants

CD (P<0.05) R L R L R L
A 0.342 0.346 C 0.342 0.346 ABC NS NS
B 0.484 0.489 AC 0.484 NS
AB 0.685 0.692 BC 0.685 0.692
Values are mean ± S.E. of three determinations
Values in parenthesis indicate specific activity (μmoles per mg of the protein) of the enzyme

with M. incognita. There was very less change in SOD was found to be higher in resistant soybean roots attacked
activity after 7 days of inoculation as compared to by M. incognita compared with uninfested roots.
uninoculated plants in both resistant as well as susceptible
genotypes. There was increase in the SOD activity in The catalase activity in the roots and leaves of
inoculated plants as compared to uninoculated plants at resistant genotypes decreased in inoculated plants as
20 days of infection. The activity of SOD from inoculated compared to uninoculated ones. However, the roots and
as well as uninoculated plants was higher jn resistant leaves of susceptible genotypes were found to contain
genotypes as compared to that of susceptible genotypes higher levels of catalase activity in inoculated plants as
at different intervals of time. Similar results were found compared to uninoculated ones at all intervals of time.
by Vanderspool et al (1994), reported that SOD activity Chen et al (1993) observed that Salicylic acid has been

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Neena Chawla et al.

found to inhibit CAT activity in many plants and thus different stages of inoculation. Mahdy and Sally (2011)
involved in plant systemic acquired resistance (SAR). reported that there was increase in the activity of both
CAT inhibition enhances the cellular level of H2O2, peroxidase and polyphenol oxidase in strawberry grown
which is presently recognized in HR, as a trigger for in the soil infested with root knot nematode when
hypersensitive cell death as well as a strong antimicrobial compared with untreated plants.
molecule (Levine et al., 1994).
The polyphenol oxidase activity was higher in the
The peroxidase activity was higher in the inoculated inoculated roots and leaves of the resistant genotypes
roots and leaves of the resistant genotypes after after 7 and 20 days of inoculation. However, in the
inoculation. However, in the susceptible genotypes, the susceptible genotypes, the roots and leaves from
roots and leaves from uninoculated plants showed higher uninoculated plants showed higher level of polyphenol
level of peroxidases as compared to inoculated plants at oxidase activity as compared to inoculated plants at

Table 4 : Polyphenol oxidase activity (PPO) (units min-1 mg-1 FW) in tomato roots (R) and leaves (L) of resistant and susceptible
genotypes against root-knot nematode

Genotype Days after inoculation

(Specific activity of PPO)
7 20
U I U I

Resistant genotypes
Hisar lalit (R) 1.20±0.06 1.98±0.12 2.42±0.18 3.24±0.26
(0.282±0.019) (0.413±0.025) (0.569±0.022) (0.713±0.024)
PNR-7 (R) 1.04±0.05 1.65±0.07 2. 12±0.16 3.01±0.24
(0.261±0.018) (0.420±0.024) (0.534±0.026) (0.601±0.027)
Hisar lalit (L) 0.88±0.03 1.21±0.09 1.39±0.06 2.98±0.18
(0.169±0.014) (0.237±0.021) (0.267±0.023) (0.552±0.027)
PNR-7 (L) 1.68±0.07 2.29±0.17 2.23±0.19 3.11±0.22
(0.286±0.019) (0.368±0.020) (0.380±0.022) (0.498±0.025)
Susceptible genotypes
Varkha Bahar 1 (R) 2.18±0.17 1.28±0.04 2.00±0.11 1.06±0.03
(0.334±0.025) (0. 199±0.024) (0.306±0.021) (0. 160±0.012)
Varkha Bahar 2 (R) 2.58±0.19 1.76±0.06 2.11±0.17 1.36±0.04
(0.363±0.025) (0.251±0.018) (0.297±0.018) (0. 179±0.016)
Varkha Bahar 1 (L) 1.83±0.04 0.98±0.02 1.01±0.03 0.65±0.02
(0.233±0.017) (0. 126±0.008) (0. 128±0.008) (0.093±0.004)
Varkha Bahar 2 (L) 1.96±0.06 1.29±0.04 1.16±0.04 0.75±0.03
(0.297±0.021) (0.161±0.010) (0. 176±0.010) (0.108±0.007)
R= Roots; L= Leaves; U= Uninoculated plants; I= Inoculated plants
CD (P<0.05) R L R L R L
A NS NS C 0.233 NS ABC NS NS
B 0.329 0.254 AC NS 0.254
AB 0.466 0.360 BC 0.466 0.360
Values are mean ± S.E. of three determinations
Values in parenthesis indicate specific activity (μmoles per mg of the protein) of the enzyme

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 Antioxidative enzymes in resistant and susceptible genotypes of tomato infected with root-knot nematode

different stages of inoculation. Rani et al (2008) found higher content of MDHAR activity in both inoculated
similar results that there was enhancement of the activities and uninoculated leaves as compared to the susceptible
of PPO enzymes in the resistant cultivars of tomato. genotypes after inoculation with M. incognita. The
DHAR activity in the roots and leaves of both inoculated
The MDHAR activity in the roots and leaves ofboth as well as uninoculated plants of resistant and susceptible
inoculated as well as uninoculated plants of resistant and genotypes increased after infection with M. incognita
susceptible genotypes increased after infection with M. at different time intervals. However, there was very less
incognita at different time intervals. However, there change in DHAR activity after inoculation in the roots of
was very less change in MDHAR activity after inoculation susceptible genotypes as compared to that in uninoculated
in the roots of susceptible genotypes as compared to that plants. El Beltagi et al (2011) observed the similar results
in uninoculated plants. The resistant genotypes recorded that the specific activity of these enzymes increased as

Table 5 : Monodehydroascorbate reductase activity (MDHAR) (μ μmoles min-1 g-1 FW) in tomato roots (R) and leaves (L) of
resistant and susceptible genotypes against root-knot nematode

Genotype Days after inoculation

(Specific activity of MDHAR)
7 20
U I U I

Resistant genotypes
Hisar lalit (R) 0.198±0.014 0.304±0.017 0.216±0.016 0.469±0.018
(0.046±0.002) (0.075±0.003) (0.051±0.004) (0.103±0.005)
PNR-7 (R) 0.324±0.015 0.722±0.022 0.421±0.018 0.848±0.020
(0.082±0.00S) (0.191±0.014) (0.106±0.009) (0.169±0.011 )
Hisar lalit (L) 0.237±0.017 0.428±0.021 0.307±0.017 0.903±0.022
(0.045±0.002) (0.084±0.005) (0.059±0.004) (0.170±0.012)
PNR-7 (L) 0.261±0.015 0.504±0.016 0.318±0.014 0.835±0.018
(0.044±0.002) (0.088±0.005) (0.054±0.003) (0.134±0.011 )
Susceptible genotypes
Varkha Bahar 1 (R) 0.209±0.013 0.241±0.021 0.243±0.017 0.270±0.018
(0.032±0.001) (0.037±0.001) (0.038±0.002) (0.040±0.003)
VarkhaBahar 2 (R) 0.185±0.016 0.225±0.019 0.211±0.017 0.254±0.019
(0.026±0.001) (0.032±0.002) (0.029±0.001) (0.034±0.002)
Varkha Bahar 1 (L) 0.167±0.015 0.206±0.019 0.198±0.013 0.240±0.017
(0.021±0.00 1) (0.026±0.002) (0.025±0.00 1) (0.030±0.002)
Varkha Bahar 2 (L) 0.219±0.015 0.301±0.021 0.279±0.016 0.317±0.018
(0.033±0.001) (0.037±0.002) (0.042±0.003) (0.044±0.003)

R= Roots; L= Leaves; U= Uninoculated plants; I= Inoculated plants

CD (P<0.05) R L R L R L
A 0.103 0.996 C 0.103 0.996 ABC 0.292 0.281
B 0.146 0.140 AC 0.146 0.140
AB 0.206 0.199 BC 0.206 0.199
Values are mean ±S.E. of three determinations
Values in parenthesis indicate specific activity (μmoles per mg of the protein) of the enzyme

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Neena Chawla et al.

μmoles min-1 g-1 FW) in tomato roots (R) and leaves (L) of resistant and
Table 6: Dehydroascorbate reductase activity (DHAR) (μ
susceptible genotypes against root-knot nematode

Genotype Days after inoculation

(Specific activity of DHAR)
7 20
U I U I

Resistant genotypes
Hisar lalit (R) 2.00±0.13 3.49±0.16 2.48±0.19 4.24±0.21
(0.470±0.015) (0.859±0.016) (0.583±0.017) (0.934±0.018)
PNR- 7 (R) 1.44±0.11 4.32±0.26 2.09±0.16 6.34±0.28
(0.363±0.015) (1.142±0.021) (0.526±0.017) (1.265±0.022)
Hisar lalit (L) 1.47±0.12 3.28±0.28 1.97±0.13 4.28±0.25
(0.282±0.022) (0.643±0.023) (0.378±0.014) (0.794±0.024)
PNR-7 (L) 2.71±0.22 3.82±0.23 3.01±0.27 4.40±0.29
(0.462±0.018) (0.673±0.023) (0.513±0.021) (0.705±0.024)
Susceptible genotypes
VarkhaBahar l (R) 2.74±0.18 2.81±0.19 3.11±0.27 3.23±0.28
(0.420±0.019) (0.427±0.022) (0.476±0.021 ) (0.482±0.022)
Varkha Bahar 2 (R) 2.04±0.14 2.26±0.16 2.99±0.23 3.14±0.24
(0.287±0.021) (0.292±0.025) (0.421±0.016) (0.428±0.019)
Varkha Bahar 1 (L) 4.16±0.25 4.32±0.26 5.03±0.26 5.19±0.27
(0.529±0.015) (0.537±0.023) (0.640±0.018) (0.651±0.022)
Varkha Bahar 2 (L) 2.48±0.16 2.61±0.17 2.85±0.24 2.97±0.26
(0.376±0.024) (0.383±0.025) (0.433±0.021) (0.444±0.023)

R= Roots; L= Leaves; U= Uninoculated plants; I= Inoculated plants

CD (P<0.05) R L R L R L
A 0.145 0.151 C 0.145 0.151 ABC 0.410 NS
B 0.205 0.214 AC 0.205 NS
AB 0.290 0.303 BC 0.290 NS
Values are mean ± S.E. of three determinations
Values in parenthesis indicate specific activity (μmoles per mg of the protein) of the enzyme

strong antioxidant defense compounds against induced change in esterase activity after inoculation in the roots
oxidative damage. and leaves of susceptible genotypes as compared to
uninoculated plants. The resistant genotypes recorded
The data on the esterase activity in the roots and higher content of esterase activity in both inoculated and
leaves of different tomato genotypes after inoculation uninoculated leaves and roots as compared to the
with M. incognita have been presented. The esterase susceptible genotypes after inoculation with M.
activity in the roots and leaves of both inoculated as well incognita. In plants with resistance to nematodes,
as uninoculated plants of resistant and susceptible esterase activity increased after nematode infection
genotypes increased after infection with M. incognita (Zacheo and Bleve-Zacheo 1995, Lambert et al., 1999
at different time intervals. However, there was very less and Andres et al., 2001).

8
 Antioxidative enzymes in resistant and susceptible genotypes of tomato infected with root-knot nematode

The native- PAGE analysis of esterase showed that Similar results were showed by Montes et al (2004) that
the roots and leaves of resistant genotypes of tomato the esterase activity increased in the resistant genotypes
plants have more intense and wider bands as compared after inoculation with root knot nematode.
to those of the susceptible genotypes after inoculation
with root knot nematode. An extra band was observed in Native- PAGE analysis of tomato plants of both
the roots and leaves of the resistant genotypes of tomato resistant and susceptible genotypes showed that the
after inoculation but there was no extra band present in isozyme bands of peroxidase were more intense and
the roots and leaves of the resistant genotypes which wider in both leaves and roots of resistant genotypes of
showed that the activity of esterase enzymes was higher tomato whereas in the susceptible genotypes the bands
in both the leaves as well as roots of resistant genotypes are less intense and wider in both roots and leaves of the
of tomato after inoculation with root knot nematode. tomato plants. Similar results were showed by Wu &

Table 7: Esterase activity (units min-1 g-1 FW) in tomato roots (R) and leaves (L) of resistant and susceptible genotypes against
root-knot nematode

Genotype Days after inoculation

(Specific activity of DHAR)
7 20
U I U I

Resistant genotypes
Hisar lalit (R) 1.87±0.17 2.66±0.23 2.11±0.19 3.43±0.27
(0.440±0.018) (0.655±0.024) (0.496±0.018) (0.756±0.028)
PNR-7 (R) 2.51±0.21 3.16±0.23 3.01±0.25 4.80±0.27
(0.632±0.021) (0.835±0.026) (0.758±0.022) (0.958±0.025)
Hisar lalit (L) 3.23±0.24 4.81±0.26 3.92±0.24 5.68±0.28
(0.621±0.026) (0.943±0.027) (0.753±0.024) (1.053±0.028)
PNR-7 (L) 2.14±0.15 3.78±0.27 2.71±0.25 4.93±0.27
(0.365±0.025) (0.667±0.027) (0.426±0.022) (0.790±0.025)
Susceptible genotypes
Varkha Bahar 1 (R) 1.09±0.07 1.13±0.09 1.69±0.06 1.72±0.08
(0.167±0.013) (0.175±0.014) (0.259±0.022) (0.267±0.024)
Varkha Bahar 2 (R) 2.12±0.15 2.25±0.21 2.73±0.23 2.85±0.26
(0.297±0.018) (0.308±0.023) (0.385±0.022) (0.390±0.025)
Varkha Bahar 1 (L) 3.19±0.25 3.29±0.27 3.92±0.32 4.08±0.33
(0.406±0.016) (0.417±0.024) (0.499±0.021) (0.512±0.022)
Varkha Bahar 2 (L) 1.96±0.14 2.07±0.16 2.21±0.17 2.34±0.19
(0.245±0.024) (0.269±0.025) (0.326±0.024) (0.335±0.025)

R= Roots; L= Leaves; U= Uninoculated plants; I= Inoculated plants

CD (P<0.05) R L R L R L
A 0.148 0.145 C 0.148 0.145 ABC 0.420 NS
B 0.210 0.205 AC 0.210 NS
AB 0.297 0.291 BC 0.297 0.291
Values are mean ± S.E. of three determinations
Values in parenthesis indicate specific activity (μmoles per mg of the protein) of the enzyme

9
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12
 Effect of VA Mycorrhiza on root diseases of tobacco
Vol. 43, No. 1, pp. 13-16 Indian Journal of Nematology June, 2013

Effect of VA Mycorrhiza on Root Disease (Pythium aphanidermatum and

Meloidogyne incognita) in Tobacco
D.V. SUBHASINI* AND S. RAMAKRISHNAN**

*Microbiology, Central Tobacco Research Institute, Rajahmundry-533 105, India

**Central Tobacco Research Institute RS, Hunsur, Karnataka, India
*Corresponding author, E-mail: dv_subhashini@[Link]

Received on 22-10-2012 and Accepted on 13-01-2013

ABSTRACT: The interaction studies between five VAM fungi, i.e., Glomus mosseae, G. fasciculatum, G. constrictum, Acaulospora laevis
and damping-off root rot pathogen (Pythium aphanidermatum) on tobacco (Nicotiana tabacum L.) seedbeds revealed that all the VAM
fungi colonized the roots efficiently within 30 days of inoculation. However, the per cent colonization was higher in the seedlings
inoculated with G. fasciculatum. The per cent disease severity in 30 day old seedlings was significantly less in treatments with VAM
fungi compared to non-mycorrhizal treatment. The impact of VA-mycorrhizal fungi i.e., Glomus mosseae, G. fasciculatum, G.
constrictum, Acaulospora laevis and root-knot nematodes on tobacco plants and the possibility of reducing the harmful effects of root-
knot nematodes using VAM fungi, was carried out in pot experiments. The VAM fungus G. fasciculatum significantly increased VAM
infection of tobacco plants infected or not infected with nematodes, whereas Meloidogyne incognita significantly reduced them,
Mycorrhizal infection percentage and number or galls were affected by variation in mycorrhizal strain. The results indicated that
the use of G. fasciculatum as a biocontrol agent is a promising technology for tobacco production.

Key words: Glomus fasciculatum, Meloidogyne incognita, root-knot nematodes and tobacco.

Damping-off is the most common and widespread effect of VAM fungi on the severity of damping off
nursery disease of tobacco (Nicotiana tabaccum L.) disease in tobacco seed beds. Root- knot disease is one
caused by soil-borne, hydrophilic fungus Pythium of the major problems effecting crop production
aphanidermatum (Edson) Fitzp. It is responsible for throughout India. The disease is caused by root-knot
poor seedling stand. Conspicuous symptom of this disease nematodes, which belong to the genus Meloidogyne.
is the sudden collapse of young seedlings in patches Among the susceptible crops the most important are
leading to uneven stand. Brown watery soft rot of young tobacco, tomato, egg plant, papaya and cotton. The
seedlings, girdling of hypocotyls and finally toppling and losses due to nematode damage in tobacco, egg plant,
death of seedlings, leading to wet rot are the characteristic cotton and coffee were 50% or more (Borah et al.,
symptoms. The pathogen spreads quickly and affects the 2009). Indirect losses associated with root-knot disease
entire seed bed causing enormous loss of seedlings. The are caused by secondary attack of other pathogens,
use of fungicides at present is unavoidable. However, inefficient utilization of fertilizers and water and high cost
chemical control of the disease is expensive and disturbs of chemical treatments. Vesicular-arbuscular mycorrhizal
soil ecology (Alagarasamy & Jeyarajan, 1989). VAM (VAM) fungi are beneficial soil fungi which form a
fungi are known to impart resistance against soil borne symbiotic association with roots of many plants. Many
plant pathogens, especially causing root rots and wilts research papers about the importance of VAM fungi in
(Moiler Kaare et al., 2009). Apart from this, the beneficial agriculture have been published (Shreenivasa et al.,
effects of vesicular-arbuscular mycorrhizal (VAM) fungi 2007).
on the growth of various crop plants have been explored
and documented in literature (Hamel & Strullu, 2006). The subject of biological antagonism in the rhizosphere
Hence, the present study was carried out to see the and root region is of long standing. The VAM fungi and

13
D.V. Subhasini and S. Ramakrishnan

root-knot nematodes are members of the microbial grain medium @ 50g/m2 nursery bed. The observations
population of the root region and they can compete with on disease severity, mycorrhizal root colonization were
each other for the same site in the rhizosphere. Hence, recorded after 30 days of pathogen [Link]
the beneficial VAM fungi might be expected to reduce weeks old mycorrhizal tobacco seedlings were
or even eliminate the harmful effects imposed by root- transplanted at the rate of one seedling per pot Mycorrhizal
knot nematodes. In plants infected with both mycorrhizal tobacco plants received Meloidogyne incognita
fungi and nematodes, mycorrhizal inoculation substantially inoculum at the rate of 2000 eggs/plant. The treatments
reduced adult nematode development (Kantharaju et al. were Nematode, Mycorrhiza, Nematode + Mycorrhiza
, 2005). Therefore, the concept of using VAM fungi as and [Link] treatment was replicated three times.
a biocontrol agents is a promising perspective of these Clay pots (30 cm in diam.) were filled with 50 kg silty soil
fungi. Objective of this work is to study the interaction of and sterilized in oven at 140°C for 2 h. Each plant
VAM fungi and root pathogens on [Link] the received about 500 ml of tap water every two days. Pots
data available on the potential of mycorrhiza in controlling were arranged in a randomized complete block design.
soil borne diseases of tobacco is meagre, the present Roots were thoroughly washed from surrounding soil
investigation to study the effects of VA mycorrhiza on and the following parameters were determined. The
host resistance is an attempt in this direction. infection percentage was determined by root technique
(Giovanetti & Mosse, 1980), (c) for assaying nematode
MATERIALS AND METHODS infection the number of galls per plant were counted.
Also, gall rating index (R.I.) from 0-5, where 0 = 0, 1= 1-
Under the present interaction studies between VAM 10; 2 = 11-30, 3 = 31-100, 4 = 100-200 and 5 = > 200 galls
and P. aphanidermatum (damping -off causing per plant was determined .
pathogen), four VAM fungi namely G. mosseae, G.
fasciculatum, G. constrictum, Acaulospora laevis RESULTS AND DISCUSSION
and damping-off root rot pathogen (P. aphanidermatum)
were used. The inoculum of these mycorrhizal endophytes The perusal of data presented in Table 1 indicates
was multiplied on Zea mays (maize) in pot culture under that all the VA- mycorrhizal fungi colonized the root
sterile conditions. tissue efficiently in the rhizosphere soil of tobacco
seedlings, and significantly reduced the severity of
The experiment was conducted during 2007-8 at damping-off disease casued by P. aphanidermatum.
Central Tobacco Research Institute nursery site. The However, percentage of disease severity and VAM
soil was analysed before starting the experiment for its colonization varied with VAM species. Amongst the
chemical characteristics such as pH-7.6, electrical various mycorrhizal endophytes tested, G. fasciculatum
conductivity-1.76 , available N-o.25, available P-0.23 showed highest root colonization (69%) followed by G.
and available K-118. mosseae and Acaulospora laevis. G. constrictum
showed minimum colonization Higher colonization with,
Tobacco seeds were sown on m2 seed- beds along G. fasciculatum may be attributed to the adaptation of
with the VA mycorrhizal inoculum. Around 50g soil VAM fungi in native soil.
including root bits containing 10-20 viable arbuscular
mycorrhizal fungal propagules / g soil were used as VAM isolate (G. fasciculatum) showed highest
inoculum and spread as a thin layer one cm below soil reduction in the damping- off severity compared to other
surface on each seed- bed. Three replications of each VAM fungi. Tobacco seedlings inoculated with this
treatment were grown for a period of 60 days with every VAM endophyte registered less disease severity
day watering up to field capacity. compared to 100% in non- mycorrhizal seedlings
(control).G. fasciculatum inoculated seedlings had
All the treatments were inoculated with propagules healthy feeder roots except some rotting of the main root,
(mycelium) of P. aphanidermatum raised on wheat where as in control plants, complete rotting of the feeder

14
 Effect of VA Mycorrhiza on root diseases of tobacco

Table 1: Mycorrhizal Root Infection and number of Pythium infected seedlings before and after inoculation of Pythium in the
soil treated with 4 mycorrhizal cultures.

Treatment Mycorrhizal root Damped off seedlings

infection% B.I.P A.I.P

G. fasciculatum 69 1.60 (1.5) 6.22 (37.6)

G. constrictum 36 3.19 (9.1) 9.52 (89.6)
G. mosseae 54 2.91 (7.4) 13.19 (172.9)
Aculospora 47 4.19 (16.5) 9.73 (93.6)
Control 0 7.96 (62.3) 15.79 (248.3)
CD (P=0.05) 0.45 0.34

Figures in parentheses are retransformed means

roots and coverage of the main root with the mycelium Table 2: Root-Knot index of tobacco plants as influenced by
of the pathogen was observed. The VAM fungus G. inoculation with 4 different strains of mycorrhiza
mosseae was the next best in disease reduction followed
Treatment Mycorrhizal root Root-knot
by Acaulospora laevis and G. constrictum. Similar infection (%) index
observation on the reduction of root infection due to
VAM inoculation in chilli seedlings has been reported by G. fasciculatum 63 3.75
Alejo-lturvide (2008). G. constrictum 38 4.87
G. mosseae 50 2.50
The G. fasciculatum successfully infected tobacco
roots forming typical VAM structures. Amongst the Aculospora 49 3.75
various mycorrhizal endophytes tested, G. fasciculatum Control 0 5.00
showed highest root colonization (63%) followed by G.
CD (P=0.05) 0.52
mosseae(50%) and Acaulospora laevis (49%). G.
constrictum (38%)showed minimum colonization.(Table-
2) Higher colonization with, G. fasciculatum may be Table 3: Root-knot index of tobacco plants as influenced by
attributed to the adaptation of VAM fungi in native soil. mycorrhizal inoculation and root-knot nematode
(Pot culture)
Root diseases caused by nematodes are influenced Treatment Mycorrhizal root Root-knot
by the mycorrhiza, less nematode galls were found in infection (%) index
most of the mycorrrhizal plants tested (Table-2). The
Nematode 0 5.00
penetration of the larvae into the roots was not inhibited
but their development within the mycorrhizal roots was Mycorrhiza 60 2.12
restricted (Subhashini & Ramakrishnan, 2011). M. Nematode + Mycorrhiza 49 2.37
incognita formed conspicuous galls on roots of tobacco
Control 0 4.12
which were significantly greater in nonmycorrhizal than
in mycorrhizal plants.(Table-3) The reduction in galls CD (P=0.05) 0.37
number caused by mycorrhizal inoculation was also
observed by Jalaluddin et al., (2008). The tolerance of et al., 2009). The tolerance of mycorrhizal plants to
mycorrhizal plants to nematode infection reported here nematode damage was attributed to changes in root
agreed with that documented in many crops such as physiology which in turn reduced nematode penetration
tomato (Shreenivasa et al., 2007) and green gram (Sorah and/or retarded adult nematode development (Grandison

15
D.V. Subhasini and S. Ramakrishnan

& Cooper, 1986). These physiological changes were Meloidogyne incognita infecting Cucumis melo.
described by Heald et al., (1989) as increased Journal of Nematology 21: 69-73.
concentration of lignin, sugar, amino acids, phenol
Jalaluddin, M., Hajra, N.B., Firoja, K. & Shahina, F. (2008).
synthesis, and ethylene production. Effect of Glomus callosum Meloidogyne incognita and
soil moisture on growth and yield of sunflower. Pakistan
To conclude with the present investigation clearly Journal of Botany 40: 391-396.
indicates the beneficial role of VA mycorrhiza in improving
the resistance of host plant both interms of damping off, Kantharaju, V., Krishnappa, K., Ravichandra, N.G. & Karuna,
root-knot nematode and substantiates the view of K. (2005). Management of root-knot-nematode
Jalaluddin et al., (2008) that these biological systems Meloidogyne incognota on tomato by using indigenous
isolates of AM fungus, Glomus fasciculatum, Indian
could be profitably exploited in the control of soil borne
Journal of Nematology 35: 32-36.
diseases.
Moiler, K., Kristensen, K., Yohalem, D. & Larsen, J. (2009).
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Canadian journal of Plant Science 86: 941-950. VAMycorrhiza and root-knot nematode on tobacco
plants: Effects of nematode inoculum density, soil texture
Heald, C.M., Burton, B.C. & Davis, R.M. (1989). Influence of and soil sterilization. Indian Journal of Nematology 41:
Glomus intraradices and soil phosphorus on 56-61.

16
 New reports of free-living marine nematodes from Indian waters
Vol. 43, No. 1, pp. 17-23 Indian Journal of Nematology June, 2013

New Reports of Free-Living Marine Nematodes, Chromadorina

granulopigmentata (Weeser, 1951) and Neochromadora poecilosomoides
(Filipjev, 1918) (Chromadorida: Chromadoridae) from Indian Waters
K.G.M.T. ANSARI*, P.S. LYLA AND S. AJMAL KHAN

Centre of Advance Study in Marine Biology, Faculty of Marine Sciences, Annamalai University, Parangipettai-608 502, Tamil Nadu, India
*Corresponding author, E-mail: [Link]@[Link]

Received on 18-12-2012 and Accepted on 02-02-2013

ABSTRACT: This paper describes new distributional records of two species of free-living nematodes belonging to the order
Chromadorida and family Chromadoridae (Chromadorina granulopigmentata (Wieser, 1951) and Neochromadora poecilosomoides
(Filipjev, 1918)) from India waters. It also provides a description for their identification based on the morphological features.

Key words: Chromadorina granulopigmentata, Neochromadora poecilosomoides, continental shelf, Indian waters

Free-living marine nematodes constitute as much as MATERIALS AND METHODS

60-90% of the benthic meiofauna (Sajan et al., 2010).
While the importance of parasitic nematodes has been Totally 35 sediment samples were collected along 6
recognized for many decades, this is not the case for transects (off Karaikkal, Parangipettai, Cheyyur, Chennai,
free-living marine nematodes, especially those of aquatic Tammenapatanam and Singarayakonda) at the depths of
environments (Heip et al., 1985). An important feature 30-50m, 51-75m, 76-100m, 101-150m, 151-175m and
of nematode population is the large number of species below 176m. In addition, due to the presence of an
present in any habitat, often an order of magnitude higher industrial cluster in Cuddalore (SIPCOT -State Industries
than for any other taxon (Schratzberger et al., 2006). Promotion Corporation of Tamil Nadu), sampling was
Order Chromadorida is the largest order in class also done here at depths of 30-50m and below 176m.
Adenophorea and family Draconematidae currently Free-living marine nematode samples were collected
includes 15 genera and 33 species and family onboard FORV (Fishery and Oceanographic Research
Epsilonematidae includes 8 genera and 11 species Vessel) “Sagar Sampada” during December, 2008
(Steyaert et al., 2005). While few studies have been (Cruise No. 260) along the southeast continental shelf of
carried out on the qualitative and quantitative aspects of India.
nematodes (Ansari et al., 1980; Harkantra et al., 1980;
Ansari & Gauns, 1996; Nanajkar & Ingole, 2007; Sajan Two sediment samples were collected using a Smith
& Damodaran, 2007; Anila Kumary, 2008), only few McIntyre grab (having a bite area of 0.2 m2) from each
studies have been carried on their taxonomy in the Indian depth. Immediately after the grab was hauled to the
waters (Timm, 1961, 1967; Sulthan Ali et al., 1998; deck, sub-samples were taken from undisturbed grab
Chinnadurai & Fernando, 2006a, 2006b). In this backdrop samples using a glass corer (having an internal diameter
the present study describes new distributional reports of of 2.5 cm and a length of 15 cm) from the middle of grab
two species of free-living nematodes namely sample (Platt & Warwick, 1983). The samples were
Chromadorina granulopigmentata (Wieser, 1951) and fixed in buffered formalin at a concentration of 4%. In
Neochromadora poecilosomoides (Filipjev, 1918) for the laboratory, samples were washed through a set of 0.5
the first time in Indian waters. mm and 0.053 mm sieves. The sediment retained in the
17
K.G.M.T. Ansari et al.

0.053 mm sieve was decanted to extract meiofauna Material examined: 7 males and 3 females collected
following the method by Higgins & Thiel (1988). Sorting from Cheyyur 30-50m, 51-75m and 101-150m depths
of meiofauna from sediment was done by flotation (18.12.2008).
technique. The efficiency of this technique is around
95% (Armenteros et al., 2008). The meiofaunal De Man ratio:
organisms were stained with Rose Bengal prior to
extraction and were sorted and enumerated under a a b c
stereomicroscope (Meiji, Japan). All the nematodes
were mounted onto glass slides, using the formalin- Male : 26±1.42 4.83±0.24 8.01±0.18
ethanol-glycerol following Vincx (1996). Identification (24.07- 28.36) (4.33-5.11) (7.62-8.32)
of nematode species was carried out to the highest Female : 25.70±1.58 4.81±0.15 7.99±0.07
taxonomic level possible using the compound microscope (24.32- 27.42) (4.68- 4.97) (7.93- 8.07)
following the standard pictorial keys of Platt & Warwick
(1988) and the NeMys Database by Steyaert et al. Description
(2005). Drawings and morphometric measurements were
made using compound microscope (Olympus CX 41 Body long and slender with the length of 0.3 - 0.6 mm
under higher magnification of 1000x attached with ocular in male and 0.4 - 0.6 mm in female and anterior tapering
micrometer) with a Camera Lucida. and posterior conico-cylindrical. Body Maximum diameter
14 – 15 µm in male and 14 – 16 µm in female. Cuticle in
RESULTS two layers, inner one being thicker than the other one.
The homogeneous cuticle with transverse rows of dots
Totally 192 species were identified along the southeast but without lateral differentiations. Lip region rounded
continental shelf of India. Among these, two species and six small rounded papillae present in apex. Four small
(Chromadorina granulopigmentata (Wieser, 1951) cephalic setae 3 - 4 µm. Silt-like amphids, 2 - 3 µm wide
and Neochromadora poecilosomoides (Filipjev, 1918)) in male and 2 - 2.5 µm wide in female. Buccal cavity with
belonging to the order Chromadorida and family a large solid dorsal tooth and two smaller equal subventral
Chromadoridae were found to be new distributional teeth. Granular pigment spots located in the anterior
reports from Indian waters. Detailed systematic account, region with two pairs of setae. Oesophagus cylindrical
materials examined (number of specimens, place, depth with well developed posterior bulb (73 - 89 µm in male
and date of collection), brief description, feeding type, and 78 - 84 µm in female). Tail conico-cylindrical with
habitat and geographical distribution besides remarks of tapering (3.2 - 4.6 anal body diameter in male and 3.9 -
the above two species are given here. 4.2 anal body diameter in female). Spicules 16 - 23µm
(1.9 anal body diameter) measured as a curve; arcuate
1. Chromadorina granulopigmentata (Wieser, 1951) with a ventral proximal swelling, rounded distal ends and
(Fig. 1 & Plate – 1 A-F) a delicate ventral ala. Gubernaculum 10 - 12 µm. A pair
of large and a pair of smaller precloacal setae present
Phylum: Nematoda Rudolphi, 1808 around the cloacal region. Ovaries paired, equal, opposed
and reflexed. Vulva present at 56- 58% of total body
Class: Adenophorea von Linstew, 1905
length (Plate 1).
Order: Chromadorida Filipjev, 1929
Feeding type: The specimens showed buccal cavity
armed with small. According to the classification of
Family: Chromadoridae Filipjev, 1917
buccal cavity by Wieser (1953), this species is an
Genus: Chromadorina Filipjev, 1918 epigrowth feeder (2A).

Species: Chromadorina granulopigmentata Habitat: Sandy sediments.

(Wieser,1951)
18
 New reports of free-living marine nematodes from Indian waters

Fig. 1. Photomicrograph (40X) of Chromadorina granulopigmentata (Wieser, 1951)

Plate 1. Chromadorina granulopigmentata (Wieser, 1951)

A) entire male B) entire female, C) male head, D) female head, E) male tail, F) female tail

19
K.G.M.T. Ansari et al.

Distribution Description

India: Cheyyur. Body long and slender with the length of 0.9 - 1.4 mm
in male and 0.8 - 1.2 mm in female and body anteriorly
Elsewhere: Southwest England (Hansson, 1998) and narrow blended and posteriorly conical with tapering.
Plymouth (Platt & Warwick, 1988). Maximum body diameter 21 – 24 µm in male and 22 – 23
µm in female. Cuticle ornamentation complex; in anterior
Remarks oesophageal region transverse rows of punctations with
lateral differentiation of two longitudinal rows of dots
The specimens examined conformed well to the beginning at base of oesophagus and continuing down
earlier description, except for the smaller body size. The length of body, terminating half way down tail. Rounded
total body length described by Platt & Warwick (1988) lip with surrounded by six short (6 – 7 µm) and four long
was 0.6 - 0.7 mm and tail length was 4 - 4.5 anal body (10 – 12 µm) cephalic setae. Somatic setae also present
diameter in male and in female these were not recorded. throughout the body. Silt-like amphids 1 – 4 µm wide in
The body length of the specimen studied at present was male and 2 – 4 µm wide in female. Buccal cavity small
found smaller being 0.3 - 0.6 mm and the tail length 3.2 and narrow with small pointed dorsal tooth and two
- 4.6 anal body diameter in male and in female 0.4 - 0.6 subventral denticles. Oesophagus cylindrical with slight
mm body length and tail length 3.9 -4.2 anal body oval posterior bulb (134 – 156 µm in male and 130 – 151
diameter This is the first record of the species from the µm in female). Tail conical (4.8 -5.1 anal body diameter
Indian waters. in male 4.9 – 5 anal body diameter in female). Spicules
13 - 16µm (0.8 - 0.9 anal body diameter), arcuate.
2. Neochromadora poecilosomoides (Filipjev, 1918) Gubernaculum curved with length of 9 – 12 µm. Eight
(Fig. 2 & Plate – 2 A-F) small squarish precloacal supplements. Ovaries paired,
equal, opposed and reflexed. Vulva present at 52 – 53
Phylum: Nematoda Rudolphi, 1808
% of body length (Plate 2).
Class: Adenophorea von Linstew, 1905
Feeding type: The specimens showed buccal cavity
Order: Chromadorida Filipjev, 1929 armed with small. According to the classification of
buccal cavity by Wieser (1953), this species is an
Family: Chromadoridae Filipjev, 1917 epigrowth feeder (2A).

Genus: Neochromadora Micoletzky, 1924 Habitat: Sandy sediments.

Species: Neochromadora poecilosomoides (Filipjev, Distribution
1918)
India: Cuddalore – SIPCOT.
Material examined: 5 males and 3 females collected
from Cuddalore - SIPCOT (19.12.2008).
Elsewhere: England (Platt & Warwick, 1988); European
De Man ratio: waters and Mediterranean (Hansson, 1998).

Remarks
a b c
Male : 44.26±1.59 7.10±0.31 10.58±0.45 The specimens examined conformed well to the
(42.61-46.07) (6.78- 7.55) (9.98- 11.12) earlier description, except for the larger body size. The
total body length described by Platt & Warwick (1988)
Female : 43.14±0.56 6.89±0.11 10.29±0.27 was 1 - 1.1 mm and tail length was 5 - 5.7 anal body
(42.57-43.72) (6.64- 6.99) (9.96-10.46) diameter in male and in female these were not recorded.

20
 New reports of free-living marine nematodes from Indian waters

Fig. 2. Photomicrograph (40X) of Neochromadora poecilosomoides (Filipjev, 1918)

Plate 2. Neochromadora poecilosomoides (Filipjev, 1918)

A) entire male B) entire female, C) male head, D) female head, E) male tail, F) female tail

21
K.G.M.T. Ansari et al.

The body length of the specimen studied at present was Harkantra, S.N., Nair, A., Ansari, Z.A. & Parulekar, A.H.
found larger being 0.9 - 1.4 mm and the tail length 4.8 - (1980). Benthos of the shelf region along the west coast
of India. Indian Journal of Marine Science 9: 106-110.
5.1 anal body diameter in male and in female 0.8 - 1.2 mm
body length and tail length 4.9 -5 anal body diameter. This Heip, C., Vincx, M. & Vranken, G. (1985). The ecology of
is the first record of the species from the Indian waters. marine nematodes. Oceanography and Marine Biology:
An Annual Review 23: 399-489.
ACKNOWLEDGEMENTS
Higgins, R.P. & Thiel, H. (1988). Introduction to the Study of
The authors are thankful to Prof. T. Balasubramanian, Meiofauna, 488. Smithsonian Institute Press,
Dean, for the encouragement and the University Washington, DC.
authorities for the facilities. We would like to thank to an
Nanajkar, M.R. & Ingole, B.S. (2007). Nematode species
anonymous referees for comments which helped to
diversity as indicator of stressed benthic environment
improve the manuscript. The authors are also thankful to along the central west coast of India. In: Desai P.V. &
the Centre for Marine Living Resources and Ecology, Roy R. (Eds.), Diversity and life processes from ocean
Ministry of Earth Sciences, Government of India, Kochi, and land. Goa University, India, pp. 42-52.
for the financial assistance through the research project
“Marine Benthos of Indian EEZ”. Platt, H.M. & Warwick, R.M. (1983). Free living marine
nematodes. Part I. British Enoplids. Pictorial key to
world genera and notes for the identification of British
REFERENCES species. In: Kermack, D.M. & Barnes, R.S.K. (Eds.),
Synopses of the British Fauna No. 28, Cambridge
Anila Kumary, K.S. (2008). Diversity of meiobenthic nematodes University Press, Cambridge, pp. 307.
in the Poonthura estuary (Southwest coast of India).
Journal of Marine Biological Association India 50: 23- Platt, H.M. & Warwick, R.M. (1988). Free living marine
28. nematodes. Part II. British Chromadorids. Pictorial key
to world genera and notes for the identification of British
Ansari, Z.A. & Ganus, M.U. (1996). A quantitative analysis of species. In: Kermack, D.M. & Barnes, R.S.K. (Eds.),
fine scale distribution of intertidal meiofauna in response Synopses of the British Fauna No. 38, Brill. Backhuys,
to food resources. Indian Journal of Marine Science Leiden, pp. 502.
25: 259-263.
Sajan S. & Damodaran, R. (2007). Faunal composition of
Ansari, Z.A., Parulekar, A.H. & Jagtap, T.G. (1980). meiobenthos from the shelf region off west coast of
Distribution of sub-littoral meiobenthos off Goa coast, India. Journal of Marine Biological Association India
India. Hydrobiologia 74: 209-214. 49: 19-26.

Armenteros, M., Perez-Garcýa, J.A., Perez-Angulo, A., Sajan S., Joydas, T.V. & Damodaran, R. (2010). Meiofauna of
Williams, J.P. (2008). Efficiency of extraction of the western continental shelf of India, Arabian Sea.
meiofauna from sandy and muddy marine sediments. Estuarine, Coastal and Shelf Science 86: 665-674.
Revista de Investigaciones Marinas 29: 113-118.
Schratzberger, M., Warr, K., & Rogers, S.I. (2006). Patterns
Chinnadurai, G. & Fernando, O.J. (2006a). New records of of nematode population in the southwestern North Sea
free-living marine nematodes from India. Records of the and their like to other components of benthic fauna.
Zoological Survey of India 106: 45-54. Journal of Sea Research 55: 113-127.

Chinnadurai, G. & Fernando, O.J. (2006b). New records of Steyaert, M., Deprez, T., Raes, M., Bezerra, T., Demesel, I.,
free-living marine nematodes from on artificial mangrove Derycke, S., Desmet, G., Fonseca, G., de Assunc ão
of India. Journal of Marine Biological Association Franco, M., Gheskiere, T., Hoste, E., Ingels, J., Moens,
India 48: 105-107. T., Vanaverbeke, J., VanGaever, S., Vanhove, S.,
Vanreusel, A., Verschelde, D. & Vincx, M. (2005).
Hansson, H.C. (1998). North East Atlantic Taxa (NEAT): Electronic Key to the free-living marine Nematodes.
Scandinavian Marine nematoda chick-list, Tjarn [Link]
Marine Biological laboratory, pp. 37.

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 New reports of free-living marine nematodes from Indian waters

Sultan Ali, M.A., Ajmal Khan, S. & Balasubramanian, T. Vincx, M. (1996). Meiofauna in marine and brackish water
(1998). Nematodes of the Pichavaram mangroves. GIS sediments. In: Hall, G.S. (Ed.), Methods for Examination
Based Information System for Pitchavaram Mangroves of Organismal Diversity in Soils and Sediments, CAB
Sponsored by DOD/ICMAM. Centre of Advanced International, Cambridge, pp. 187-195.
Study in Marine Biology, Annamalai University, pp. 47.
Wieser, W. (1953). Die beziehungen zwischen
Timm, R.W. (1961). The marine nematodes of the Bay of mundho¨hlengestalt, erna¨hrungsweise und vorkommen
Bengal. Proceedings of the Pakistan Academy of Science bei freilebenden marinen nematoden. Arkive Fur Zoologi
1: 1-84. 2: 439-484. (In German).

Timm, R.W. (1967). Some estuarine nematodes from the

Sunderbans. Proceedings of the Pakistan Academy of
Science 4: 1-14.

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T.I. Olabiyi
Vol. 43, No. 1, pp. 24-28 Indian Journal of Nematology June, 2013

Chemo-Therapeutic and Nematoxic Effect of Composts on the Survival of

Meloidogyne hapla Juveniles
T.I. OLABIYI*

Department of Crop and Environmental Protection, Faculty of Agricultural Sciences,

Ladoke Akintola University of Technology, P.M.B. 4000, Ogbomoso, Nigeria
*Corresponding author, E-mail: [Link]@[Link]

Received on 21-12-2012 and Accepted on 02-02-2013

ABSTRACT: Aqueous extract from five different composts (Terra Ecosystem, wyvern waste, garden waste, farm yard manure and
green waste) were assessed for their effects on juvenile survival of Meloidogyne hapla in vitro. It was found that graded extracts (5,
10, 15 and 20% w/v) of the composts effectively killed M. hapla juveniles. At the 8th day exposure time,M. hapla juveniles in Terra
Ecosystem, wyvern waste, garden waste and green waste were dead, whereas few M. hapla were alive in the farm yard manure aqueous
extract till day 10. The in vitro assessment shows that Terra Ecosystem, wyvern waste, garden waste and green waste were significantly
toxic to M. [Link]-chemical analysis of the compost revealed that Terra Ecosystem contained sterols and flavonoids; wyvern
waste contained sterols, glycosides and flavonoids; garden waste contained only flavonoids; farm yard manure contained saponins,
sterols, glycosides and flavonoids; and green waste contained saponins, sterols and flavonoids.

Key words: Compost, nematode,Meloidogynehapla, Chemo-therapeutic, nematoxic

The ten most important nematode genera include research on measures that could have no or low
Meloidogyne, Pratylenchus, Heterodera, environmental impact, alternatives to synthetic pesticides,
Ditylenchus, Globodera, Tylenchulus, Xiphinema, had received very strong impulse and considered a wide
Radopholus, RotylenchulusandHelicotylenchus range options. One of the options is the application of
(Sasser & Freckman, 1987). Root knot nematodes, compost in agricultural crop production. Application of
Meloidogyne species, are economically important compost, as soil amendment or aqueous plant extract,
pathogens worldwide. They are obligate parasites infecting had caused significant reduction in population of root
plant species, including major food crops, vegetables, knot nematode, Meloidogyne spp. and reduced the
fruits and ornamental crops. They reduce the yield and nematode damage which had resulted to improve plant
quality of susceptible crops (Jonathan & Hedwig, 1991; growth and yields (Verma, et al., 1997; Ribeiro, et al.,
Yousef & S’Jacob, 1994; Olabiyi, 2008). 1998; D’Addabbo, et al., 2000).

There are various and recent European Legislations Recently, the safety and health of food have become
which restricted and revised the use of chemical pesticides major concern, due to indiscriminate use of pesticides for
on agricultural crops (Reg. CE 396/2005; 1095/2007; 33 food production and its resultant negative impacts on
and 299/2008 and 1107/2009). Agricultural crop human health, livestock, soil and environment (Primentel,
consumers are poisoned and in most cases, the prices of 2005). For this reason, farmers in the environmental
synthetic nematicides are very high and not always conscious world have started replacing synthetic fertilizers
available for the users at the time of need (Thomas, with compost. Composts have been reported to augment
1996). All these spurred researchers/ scientists globally, soil nutrient and suppress soil inhabiting nematodes
to find alternative strategies that are environmentally (Olabiyi,et al. 2007).
friendly and economically convenient. Therefore,

24
 Chemo-therapeutic and nematoxic effect of composts on the survival of Meloidogyne hapla juveniles

Decomposed plant materials have been reported to approximately 100 M. hapla population was added. The
be toxic to nematodes and have been effectively used as Petri dish containing only 1ml juvenile suspension
nematode pest control measure. These composted plant containing approximately 100 M. hapla juveniles in 20 ml
materials include wild sunflower, neem cake, neemleaf, distil water only, served as the control. Each treatment,
maize stover and cassava peel (Egunjobi and Onayemi, including the control, was replicated five times. The
1981; Riga and Lazarovits, 2001; Abbasi, et al. 2005; experimental design was complete randomized design.
Olabiyi,et al. 2007). The objective of this study is therefore A count of live nematodes was made and cumulative
to assess the potential of different organic composts on numbers of dead nematodes were put on record on daily
the juvenile survival of root knot basis for a period of 10 consecutive days.
nematode,Meloidogynehapla and also to determine the
phyto-chemical constituents of the compost. Phyto-chemical analysis

MATERIALS AND METHODS Two-hundred and fifty gram (250gm) of each

compost (Terra ecosystem, Wyvern waste, Green waste,
Preparation of compost solution: Terra ecosystem, Farm yard manure and Garden waste) was ground using
Wyvern waste, Green waste, Farm yard manure and Glen Creston Rotary hammer mill, to pass a 1mm mesh.
Garden waste were ground (2mm) using attrition mill in The samples were then stored in re-sealable polythene
the Environmental Science Laboratory of the Coventry bags. An hundred gram (100g) of the compost was
University, UK. Ten (10) ml each of the air-dried weighed separately with sensitive electric weighing
compost was transferred to plastic poly-bottles to which balance (four digital) into 500 ml ethanol (100%) and
were added 25ml of deionised water. The bottles were soaked for 3 days in a closely air-tight one (1) litre sized
capped and then shaken for 15 minutes on a reciprocating measuring cylinder. The content was filtered using
shaker (200 – 250 hub/min) and the resulting suspension Whatman filter paper number 1. The filtrate (ethanol
was filtered through Whatman filter paper number 1. extract) was used for the chemotherapeutic analysis.

Preparation of the test organism (root knot Flavonoids

nematode): Tomato (Lycopersicumesculentus L. Mill),
was grown in pot in the greenhouse and inoculated with (i) Aliquot of 4 ml of aqueous NaOH was added to 2ml
stock culture of Meloidogynehapla. Ten weeks after of each of ethanol extract. If a yellow precipitate
planting (10 WAP), eggs of Meloidogynehapla were was observed, it indicates the presence of flavonoids
extracted, using sodium hypochlorite (Hussey and Baker, in the extracts. But if otherwise, it indicates the
1973) from tomato galled roots. The eggs were incubated absence of flavonoids
at temperature of 10 - 15oC for 24 hours. The freshly
hatched second stage nematode juveniles were collected. (ii) Shinoda test: A little amount of magnesium powder
Juveniles of M. hapla in the distil water suspension was and 3 drops of concentrated HCl were added to 4 ml
concentrated and standardized so that each 1ml of each of the ethanol extract. If a red colour was
suspension contained approximately 100 juveniles. observed, it indicates the presence of flavonoids. If
otherwise, it shows the absence of flavonoids
Effect of different compost on M. hapla juveniles
(nematode bio-assay): Different compost solutions Sterols
were prepared into 1, 5, 10,15% and 20% w/v
concentrations in the Environmental Science Laboratory, Aliquot of 2ml concentrated H2SO4 was added to 1
Coventry University, UK. Aliquot of 20 ml of each ml of each ethanol extract. If a brownish red colour was
compost solution was dispensed into Petri-dish and observed, it shows the presence of sterols. But, if
thereafter, 1 ml each of nematode suspension containing otherwise, it shows the absence of sterols.

25
T.I. Olabiyi

Glycosides Test for Saponins

Aliquot of 10 ml of 50% H2SO4 was added to 1 ml Frothing test: Aliquot of 2ml of each ethanol extract
of each ethanol extract. The mixture was heated on was shaken vigorously in the test tube for 2 minutes,
water bath for 15 min. Then, 10ml of fehling solution frothing shows the presence of saponins. But if otherwise,
(5cm3 each of fehling solutions A and B) was added and it indicates absence of saponins.
boiled. If a red precipitate was observed it shows the
presence of glycoside. But if otherwise, it shows the Emulsion test: Aliquot of 5 drops of olive oil was added
absence of glycosides. to 3 ml of the weed extract in test tube and mixture was
vigorously shaken. If a stable emulsion is formed, it
Tannins indicates the absence of saponins. But if froth is formed,
it indicates presence of saponins.
To 2ml of each of the ethanol extract was added 5
drops of Fe2Cl3 solution. If a dirty green precipitate was RESULTS AND DISCUSSION
observed, it indicates the presence of tannins. But if
otherwise, it indicates the absence of tannins. Table 1 shows the survival potential of
Meloidogynehapla in different compost concentrations

Table 1: Survival of M. hapla in different compost concentrations

Different compost Number of live nematodes at day

Concentrations (% w/v) 1 2 3 4 5 6 7 8 9 10

Terra ecosystem 5 100 52e 38b 22b 7a 4a 3a 0a0a0a

10 100 47d 31b 20b 9a 4a 1a 0a0a0a
15 100 40d 25b 11a 4a 2a 1a 0a0a0a
20 100 26bc 18a 8a 3a 1a 0a0a0a0a
Wyvern waste 5 100 32c 14a 10a 7a 1a 0a0a0a0a
10 100 28c 13a 11a 9a a a a a a
00000
15 100 20b 10a 8a 7a 1a 0a0a0a0a
20 100 15a 12a 10a 8a 1a 0a0a0a0a
Garden waste 5 100 24b 20a 13a 12a 4a 1a 0a0a0a
10 100 24b 17a 7a 5a 2a 1a 0a0a0a
15 100 20a 14a 7a 4a 1a 0a0a0a0a
20 100 21b 12a 6a 5a 0a0a0a0a0a
Farm yard manure 5 100 85g 72c 38c 28b 26b 24b 24b 22b 20b
10 100 80g 66c 34c 22b 21b 20b 20b 19b 17b
15 100 88g 68c 33c 25b 22b 20b 20b 18b 17b
20 100 76f 61c 35c 28b 25b 21b 20b 20b 18b
Green waste 5 100 21b 14a 10a 9a 3a 0a0a0a0a
10 100 18a 16a 11a 7a 3a 1a 0a0a0a
15 100 16a 14a 13a 13a 4a 0a0a0a0a
20 100 13a 13a 12a 11a 1a 0a0a0a0a
Control (ddH2O) NS 100 100h 94d 91d 90c 85c 81c 80c 71c 65c

NS = Not statistically significant

Means followed by the same letter along the same column are not statistically different at 5% probability

26
 Chemo-therapeutic and nematoxic effect of composts on the survival of Meloidogyne hapla juveniles

Table 2: Phyto-chemical analysis of different composts

Saponins Tannins Sterols Glycosides Flavonoids

Terra ecosystem A A P A P
Wyvern waste A A P P P
Green waste P A P A P
Farm yard manure P A P P P
Garden waste A A A A P

Key: P = Present A = Absent

within 10 days. It was observed that compost have lethal extracts (100% concentrations) of Lactua sativa,
effect on M. hapla and the toxic effects are days to Amnimajus, Artemisia pallens and Artemisia annua
exposure and level of concentration dependent. However, resulted in 100% Meloidogyne incognita mortality
toxic potential of compost differ from one type of within 24 hours of exposure in vitro (Pandey, 1990).
compost to the other. By day 8 of exposure, all nematodes
in Terra ecosystem, Wyvern waste, Garden waste and The result of phyto-chemical analysis revealed that
Green waste were dead, whereas few were still found the compost contained saponins, tannins, sterols,
living in the Farm yard manure, even till after day 10. glycosides and flavonoids. Several plants and compost
Though there were reduction in the number of live are known for their toxicants and metabolites. Natural
nematodes in the control (deionised distilled water), but plant products have the ability to produce environmentally
live nematodes recorded, showing that deionised distilled friendly bio-toxicants that are bio-degradable and
water does not have toxic effect on M. hapla. efficacious against pathogenic organisms, including soil
inhabiting plant parasitic nematodes (Schmutterer, 1990;
Table 2 shows the chemotherapeutic potentials of Jackai, et al., 1992).Brimstone, Morindalucida,
the different composts. Presence of saponins, tannins, contained saponins and flavonoids which were reported
sterols, glycosides and flavonoids were examined on to have bio-nematicidal properties (Olabiyi,et al., 2008).
each of the compost. The results elicit the presence of
saponins in green waste and farm yard manure; none of CONCLUSION
the compost had tannins; terra ecosystem, wyvern waste,
green waste and farm yard manure showed the presence The current research indicates that compost contained
of sterols and flavonoids; while only wyvern waste and certain bio-chemical which was proved to be toxic to
farm yard manure had glycosides. Garden waste did not Meloidogyne hapla in the laboratory. However, there
contain any of the tested chemical compounds. is need to do further investigation on the field.

Aqueous extract from Terra Ecosystem, wyvern ACKNOWLEDGEMENT

waste, garden waste, farm yard manure and green waste
compost were assessed for their effects on juvenile Many thanks to Professor P.J.C. Harris of Faculty
survival of Meloidogyne hapla in vitro. The tested of Business, Environment and Society, Coventry
composts significantly reduced the juvenile survival rate University, Coventry, United Kingdom for the given
of M. hapla compared to the [Link] result research opportunities and financial support. My
corroborates the earlier findings of Sasanelli and Addabbo appreciation also goes to Neil Thompson, Sue Tompsett
(1993), Pandey et al. (1997) and Olabiyi et al. (2007) and Richard Collins for the laboratory assistance at the
who reported that organic manure significantly reduced Environmental Science Laboratory, Coventry University,
the population of Meloidogyne incognita. Aqueous United Kingdom during the course of this research.

27
T.I. Olabiyi

REFERENCES International Nematology Network Newsletter 7: 19-

20.
Abbasi, P.A., Riga, E., Conn, K.L. & Lazarovits, G. (2005).
Effects of neem cake soil amendment on reduction of Pandey, R., Singh, H.B. & Gupta, M.L. (1997). Antagonistic
damping off severity and population densities of plant impact of vesicular-arbuscularmychorrhizal fungi on
parasitic nematodes and soil borne plant pathogens. Meloidogyne incognita population development in
Canadian Journal of Plant Pathology 27: 38-45. Japanese [Link] Journal of Tropical Plant
Disease 15: 237-245.
D’addabbo, T., Sasanelli, N., Lamberti, F., Greco, P. & Carella,
A. (2000).Control of root knot nematode by olive and Pimentel, D., Hepperly, P., Hanson, J., Douds, D. & Seidel, R.
grape pomace soil amendment. In: Gullino, M.L., Katan, (2005). Environmental, energetic and economic
J. andMatta, A. (Eds). Proceedings of the fifth comparison of organic and conventional farming
international symposium on chemical and non- systems. Bio Sciences 55: 573-582.
chemical soil and substrate disinfestation. Acta
Horticulture 523: 53-55. Ribeiro, R.C., Mizobutsi, E.H., Silva, D.G., Pereira, J.C.R. &
Zambolim, L. (1998). Control of Meloidogyne javanica
Egunjobi, O.A. & Onayemi, S.O. (1981). The efficacy of water on lettuce with organic amendments. Fitopatologia
extract of neem (Azadirachtaindica L.) leaves as a Brasileira 23: 42-44.
systemic nematicide. Nigerian Journal of Plant
Protection 5: 70-74. Riga, E. & Lazarovits, G. (2001). Development of an organic
pesticide based on neem tree products. American
Hussey, R.S. & Baker, K.R. (1973).A comparison of methods Phytopathological Society/ Mycological Society of
of collecting inocula for Meloidogyne species including America/ Society of Nematology Joint meeting Abstract
a new technique. Plant Disease Report 57: 1025-1028. of Presentations. Salt Lake City, Utah. Publication no:
p.2001-0096-SON. Phytopathology 91: 141.
Jackai, L.W.N., Inang, E.E. & Nwobi, P. (1992).The potential
for controlling post flowering pests of cowpea using Sasanelli, N. & Addabbo, T.D. (1993).Potential application of
Neem, Azadirachtaindica. A. Juss. Tropical Pest the leaves of Rutagraveolens for controlling
Management 38: 56-60. Meloidogynejavanica on sunflower. Russian Journal
of Nematology 1: 117-120.
Jonathan, D.E. & Hedwig, H.T. (1991).Root knot nematode,
Meloidogyne species and races. Manual of Agricultural Sasser, J.N. & Freckman, D.W. (1987). A world perspective
Nematology (Eds. William, R. and Nickel, 1991), Mercel on Nematology: The role of society. Vistas on
Dekker Inc. pp. 191-192. Nematology. In: J.A. Veech and D.W. Dickson (Eds), pp.
7-14.
Olabiyi, T.I. (2008). Pathogenicity study and nematoxic
properties of some plant extracts on the root-knot Schmutterer, H. (1990). Properties and potential of natural
nematode pest of Tomato, Lycopersiconesculentum pesticides for the neem tree, Azadirachta indica. Annual
(L.) Mill. Plant Pathology Journal 7: 45-49. Review Entomology 35: 271-298.

Olabiyi, T.I., Akanbi, W.B. & Adepoju, I.O. (2007).Control of Thomas, W.B. (1996). Methly bromide: Effective pest
certain nematode pests with different organic manure on management tool and environmental threat. Journal of
[Link]-Eurasian Journal of Agricultural Nematology (Supplement) 28: 586-590.
and Environmental Sciences 2: 318-320.
Verma, R.D., Mahendra, S., Samar, R. & Sharma, G.L. (1997).
Olabiyi, T.I., Oyedunmade, E.E.A. & Ibikunle, G.J. (2008). Effect of soil amendment against root knot nematode
Phytochemical screening and nematoxic effect of Meloidogyne incognita on bottle gourd. Indian Journal
Brimstome, Morindalucida, on nematode pests of of Nematology 27: 255-256.
Amaranth, Celosia argentea. Biological Agriculture
and Horticulture 26: 131-137. Yousef, D.M. and S’Jacob, J.J. (1994).A nematode survey of
vegetable crops and some orchards in the Ghor of
Pandey, R. (1990). Studies on the phytonematoxic properties Jordan. Nematologia Mediterranea 22: 11-15.
in the extract of some medicinal and aromatic plants.

28
 Efficacy of wild sunflower compost on root lesion nematode
Vol. 43, No. 1, pp. 29-33 Indian Journal of Nematology June, 2013

Efficacy of Wild Sunflower Compost on Root Lesion Nematode,

Pest of Maize
T.I. OLABIYI1*, T.A. OGUNNIRAN1, O.J. OJO1, J.J. ATUNGWU2 AND S.A. ABOLUSORO3

1
Department of Crop and Environmental Protection, Faculty of Agricultural Sciences,
Ladoke Akintola University of Technology, P.M.B. 4000, Ogbomoso, Nigeria
2
Department of Crop Protection, Federal Unversity of Agriculture, P.M.B. 2240, Abeokuta, Nigeria
3
Department of Crop Production Technology, DAC/ABU Kabba, Kogi State, Nigeria
*Corresponding author, E-mail: [Link]@[Link]

Received on 21-12-2012 and Accepted on 06-02-2013

ABSTRACT: Field trials were conducted during 2010 and 2011 planting seasons to assess the bio-nematicidal potential of wild
sunflower (Tithonia diversifolia) compost in the control of Pratylenchus brachyurus infection on maize (Zea mays) cv ACR 97. Field trials
were arranged in randomized complete block design, with 5 treatments, each replicated 4 times. Experimental plots where no
compost were applied served as the control; while other treatments were 1.0, 1.5, 2.0 and 2.5 tons/ha. The result obtained shows that
higher application rates of 2.0 and 2.5 tons/ha. gave significant (p<0.05) control measure on P. brachyurus with resultant higher growth
and yield of maize. The soil nematode population where 2.0 and 2.5 tons/ha were applied had significant (p<0.05) lower number of
P. brachyurus. The result of the phytochemical screening revealed the presence of tannins, saponins, alkaloids and flavonoids, which
were hypothesized to be the nematoxic principles in wild sunflower compost.

Key words: Compost, Pratylenchus brachyurus, Zea mays, sunflower

Maize (Zea mays L.) belongs to the family Graminae. young seedlings may be observed in cases of heavy
Maize is widely cultivated throughout the world and it is infestation (Egunjobi and Ekundare, 1981).
one of the most important cereal grains in the world,
providing nutrients for humans and animals. It also The use of chemicals has been found to be the most
serves as a basic raw material for the production of effective method of controlling pests on Agricultural
starch, oil, protein, alcoholic beverages foods, and fuel. crops. However, the prohibitive costs as well as hazards
Maize has very high average yield per hectare when associated with their application have greatly reduced
grown under optimum condition. Despite the wide spread their use by many farmers (Olabiyi, 2004). Today the
use of maize, its production and cultivation has been world is becoming more and more aware of the
greatly reduced by insect pests, diseases and soil inhabiting environmental dangers posed by chemical pesticides.
pathogens that affect the growth and yield of maize on Even in the advanced countries where these chemicals
the field. are produced and farmers have sufficient skill for their
application, synthetic chemicals are now being replaced
Root lesion nematodes, Pratylenchus brachyurus with other environmentally friendly methods of pest and
Filipjev, are the major species of nematodes found on disease control for various reasons including water and
maize and are mainly root parasites. The below ground atmospheric pollution which continue to render the world
symptoms on the roots are the presence of lesions/ agricultural foods unsafe for mankind (Egunjobi and
blisters on the root, excessive branching of root, and Afolabi, 1976; Thomas, 1996; Olabiyi, 2005).
shortening of the root. The above ground symptoms
manifested by affected crops are dwarfing, patchy Organic manure when incorporated in large amount
growth and low yield. Poor germination and death of into the soil reduces the population of the soil inhabiting

29
T.I. Olabiyi et al.

pest and pathogen. Wild sunflower used as soil amendment Phyto-Chemical analysis: The presence of tannins,
is very promising for nematode control (Olabiyi et al. saponins, flavonoids, alkaloids, glycosides and sterols in
2007). The use of compost in crop protection when fully wild sunflower compost was assessed at the University
developed or integrated will enhance safe food crop Central Research Laboratory, Ladoke Akintola University
production, reduce environmental pollution and also make of Technology, Ogbomoso, Nigeria.
our agricultural produce to gain international recognition.
The objective of this paper is therefore to assess the Test for Saponin (frothing test): 2 ml of crude extract
effect of wild sunflower (Tithonia diversifolia) compost of wild sunflower compost in a test tube was vigorously
on root lesion nematode (Pratylenchus brachyurus) shaken for 2 min. Frothing in the extract indicates the
pest of maize (Zea mays). presence of saponins.

MATERIALS AND METHODS Test for Saponin (emulsion test): Five drops of olive
oil were added to 3 ml of the extract of wild sunflower
Preparation of compost materials compost in a test tube and the mixture were vigorously
shaken. A stable emulsion formed in the extract indicates
A windrow method was used for the preparation of the presence of saponins.
compost at the Teaching and Research farm, Ladoke
Akintola University of Technology, Ogbomoso, Nigeria. Test for Flavonoids (Shinoda test): 1 ml of 10%
Freshly harvested wild sunflower (100 kg) was chopped NaOH was added to 4 ml of extracts of wild sunflower
(5-10 cm sizes), wrapped with plastic and properly compost. A yellow colour observed indicates the presence
covered up for a period of 5 months with regular turning of flavonoids.
(monthly interval) with garden spade. The fully
decomposed materials (compost) were spread and air Test for tannins: Two drops of 5% ferric chloride were
dried. The compost, in air-dried form, was ground into added to 1 ml of crude extract of wild sunflower compost.
powder form using attrition mill. An observed dirty green precipitate in the extract indicates
the presence of tannins.
Field experiment
Test for sterols: 1 ml of concentrated sulphuric acid
The experimental field of 350m2 size was ploughed (H2SO4) was added to 3 ml of crude extract of wild
twice and partitioned into plots. Five ridges of 4m2 were sunflower compost. A colourless solution observed
constructed within plots. The field experiment was indicates the absence of sterols.
carried out at the Organic Garden section, Teaching and
Research Farm, Ladoke Akintola University of Test for glycosides: 10 ml of 50% concentrated
Technology, Ogbomoso, Nigeria in 2009 and 2010. The sulphuric acid (H2SO4) was added to 1 ml of crude
experimental design was randomized complete block extract of wild sunflower compost in a test tube. The
design, with 5 treatments replicated 4 times. Three seeds mixtures were heated in boiling water for 15 min.
were planted per stand which was later thinned down to Fehling’s solution (5 ml each of Fehling’s solution A and
one healthy maize plant per stand with an average B) were added and the mixture was boiled. A colourless
distance of 25cm intra-row on the ridges. Each of the solution observed indicates the absence of glycosides.
maize stands was inoculated two weeks after planting
with 100g P. brachyurus infected maize root. The Test for alkaloids: 1 ml of 1% hydrochloric (HCl) acid
compost, in power form, was applied as soil amendments was added to 3 ml of crude extract of wild sunflower
in ring form, 2-3cm away from maize plant and at the compost in the test tube. The mixture was boiled for 20
rates of 1.0, 1.5, 2.0 and 2.5tons/ha. The experimental min and filtered. Portions of the filtrate were treated with
plots where no compost was applied served as the two drops each of Mayer’s and Wagner’s reagent
control. separately. An observed creamy white (Mayer’s reagent)

30
 Efficacy of wild sunflower compost on root lesion nematode

and reddish brown (Wagner’s reagent) precipitates The result presented on Table 2 elicits the effects of
indicates the presence of alkaloids. application rates of wild sunflower compost on the yield
of maize. While there was no significant difference
Data collection between the treated maize and control with respect to
number of cob per plant, there were significant differences
Data were collected on plant height, number of leaf on the number of grain per cob and fresh cob weight in
per plant, plant girth, number of cob per plant, number of view of the maize plants that were treated and those that
grain per cob, fresh cob weight, population of root lesion were not treated (control). Maize plants that were
nematode in 10g root sample at both years were pooled treated with wild sunflower compost have higher number
together, and thereafter subjected to analysis of variance of grains per cob and fresh cob weight than the untreated
(ANOVA) and means were separated using Duncan maize (control). Maize plants treated with wild sunflower
Multiple Range Test (DMRT) at 5% probability level. compost of between 1.5 and 2.5 tonnes per hectare
Data were also collected on the phyto-chemical analysis performed significantly better than those maize plants
of wild sunflower compost. treated with 1.0 tonne per hectare wild sunflower compost
and control.
RESULTS AND DISCUSSION
Table 2: Effects of wild sunflower compost on the yield of
Two years field trials were carried out during 2010 maize infected with P. brachyurus
and 2011 planting seasons in order to assess the bio-
nematicidal potentials of wild sunflower compost in the Treatments Mean number Mean number Fresh cob
control of maize root lesion nematode, Pratylenchus of cob/ plant of grain/ cob weight (kg)
brachyurus. The effects of wild sunflower compost on Control 1 105.4d 0.06c
the growth of maize infected with P. brachyurus were
presented on Table 1. It was evident that maize treated 1.0 tonne/ ha. 1 311.3c 0.09b
with wild sunflower compost significantly have higher 1.5 tonne/ ha. 1 412.6b 0.14a
plant height, number of leaves per plant and plant girth 2.0 tonnes/ ha. 1 582.1a 0.15a
than the control. Maize plants that were treated with wild
2.5 tonnes/ ha. 1 581.5a 0.16a
sunflower compost of between 1.5 and 2.5 tonnes per
hectare performed better than those treated with 1.0 Means followed by the same letter(s) along the same column
tonne per hectare and control. are not significantly different at 5% probability level according
to Duncan multiple range tests (DMRT).
Table 1: Effects of wild sunflower compost on the growth of
maize infected with P. brachyurus Table 3 shows the effect of different application
rates of wild sunflower compost on the population dynamics
Treatments Mean number Mean plant Plant girth
of P. brachyurus in the root of maize. Maize plants that
of leaf/plant height (cm) (cm)
were treated with wild sunflower compost significantly
Control 9.3b 65.4d 2.0b have lower P. brachyurus population in the root (10g) of
1.0 tonne/ ha. 9.8b 73.3c 2.8a maize.
1.5 tonne/ ha. 10.7a 119.6b 3.0a Phyto-chemical components of wild sunflower
2.0 tonnes/ ha. 13.0a 152.1a 4.0a compost were presented on Table 4. It was evident that
2.5 tonnes/ ha. 13.1a 151.5a 4.0a wild sunflower compost contains tannins, saponins,
flavonoids and alkaloids while it lacks sterols and
Means followed by the same letter(s) along the same column glycosides. The presence of flavonoids in the sunflower
are not significantly different at 5% probability level according
compost might be the reason why it gave effective
to Duncan multiple range tests (DMRT).
control measure on maize.

31
T.I. Olabiyi et al.

Table 3: Effects of wild sunflower compost on the root lesion Table 4: Phyto-chemical analysis of wild sunflower compost
nematode population
Chemical constituents Inference
Treatments Mean root lesion nematode
in 10gm root sample Tannins Present
Saponins Present
Control 110.5b
Flavonoids Present
1.0 tonne/ ha. 32.5a
Alkaloids Present
1.5 tonne/ ha. 31.7a
Gycosides Absent
2.0 tonnes/ ha. 31.5a
Sterols Absent
2.5 tonnes/ ha. 30.5a

Means followed by the same letter(s) along the same column

are not significantly different at 5% probability level according Pratylenchus brachyurus juveniles within 6 days
to Duncan multiple range tests (DMRT). exposure at concentration of 1.5 in vitro.

In conclusion, the results of this study unveil the bio-

The results of this experiment showed that wild
nematicidal potentials of wild sunflower compost and
sunflower (Tithonia diversifolia) compost significantly
show that it could be used in the control of Pratylenchus
reduced the infection potential of Pratylenchus
brachyurus infection on maize.
brachyurus on maize. The bio-nematicidal effects of
composted materials as soil amendments with plant- and
REFERENCES
animal-based materials have been reported previously
(D’Addabbo, 1995; Akhtar and Malik, 2000; Olabiyi, Akhtar, M. & Malik, A. (2000). Roles of organic amendments
2005). Application of composts as soil amendments are soil and soil organisms in the biological control of plant-
found to improve soil quality and soil microbial population parasitic nematodes: a review. Bioresource Technology
dynamics, and also suppress the activity of plant soil- 74: 35-47.
inhabiting pathogens and pests, including plant parasitic
nematodes (Bailey and Lazarovitz, 2003; Chang et al. Bailey, K.L. & Lazarovitz, G. (2003). Suppressing soil-borne
diseases with residue management and organic
2007). Composts prepared from fresh olive pomace amendments. Soil and Tillage Research 72: 169-205.
combined with a low percentage of chicken manure;
cow manure and lettuce crop residues; municipal green Chang, E.H., Chung, R.S. & Tsai, Y.H. (2007). Effect of
wastes combined with sewage sludge; municipal green different application rtes of organic fertilizer on soil
wastes combined with soil; and mycelium waste from enzyme activity and microbial population. Soil Science
penicillin production were reported to significantly reduce and Plant Nutrition 53: 132-140.
the soil population soil inhabiting nematodes of barley
D’Addabbo, T. (1995). The nematicidal effect of organic
(Renco et al. 2010). Moreover, Olabiyi (2007) reported amendments: a reviewof the literature, 1982-1994.
reduction in number of galls and egg masses as well as Nematologica Mediterranea 23: 195-198.
increase in plant growth as a result of treating nematode
infected cowpea with composted wild sunflower. Egunjobi, O.A. & Ekundare, G.O. (1981). Cassava peelings as
a soil amendment and its effects on maize yield in soil
The phytochemical analysis revealed the presence infested with Pratylenchus brachyurus. Nigerian
of flavonoids in the wild sunflower compost. This Journal of Plant Protection 5: 80-87.
investigation supported earlier reports of the nematicidal
Olabiyi, T.I. (2004). Assessment of the nematicidal properties
properties of flavonoids by Verma et al (1978), Zavaleta- of extracts from Tagetes erecta, Ocimum gratissimum,
Mojia et al (1993) and Submaniyan and Vadivelu (1990) Hyphs suaveolens and Crotalaria retusa. Ph.D. Thesis.
who reported that Crotolaria spectabilis leaves and Department of crop production. University of Ilorin.
roots posses certain phyto-chemicals that killed Ilorin. Nigeria. 177 pp.

32
 Efficacy of wild sunflower compost on root lesion nematode

Olabiyi, T.I. (2005). Application of plant materials in the Thomas, W.B. (1996). Methyl bromide: Effective pest
control of nematode pests of tomato in rural areas of management tool and environmental threat. Journal of
Nigeria: A case study. Afro-Asian Journal of Rural Nematology (Supplements) 28: 586-590.
Development 38: 49-55.
Verma, M.K., Sharma, H.C. & Panthak, V.N. (1978). Efficiency
Renco, M., Sasanelli, N., D’Addabbo, T. & Papajova, I. (2010). of Tagetes patula and Sesamum orientate against root
Soil nematode community changes associated with knot nematodes of egg plant. Plant Diseases Reporter
compost amendments. Nematology 12: 681-692. 62: 274-275.

Subramaniyan. S. & Vadivelu, S. (1990). Effects of Crotalaria Zavaleta-Mojia, E., Castro, A.E. & Zamudis, V. (1993). Effects
spectabilis Extracts on Meloidogyne incognita. del cultivo e incorporacion de Tagetes erecta L. Sobre
International Nematology Network News 7: 8-9. lapoblacion e infecio de meloidogyne incognita (Kofoid
and white) chitwood in chilli (Capsicum anuum L.).
Nematropica 23: 49-56.

33
Prasanna Holajjer et al.
Vol. 43, No. 1, pp. 34-39 Indian Journal of Nematology June, 2013

Evaluation of Heterocystous and Non Heterocystous Cyanobacterial Species

for Nematicidal Activity
PRASANNA HOLAJJER1*, ANJU KAMRA, H.S. GAUR AND DOLLY WATTAL DHAR2

1
Crop Protection Unit, Directorate of Groundnut Research (DGR), Junagadh-362001, Gujarat, India
2
Centre for Conservation and Utilisation of Blue Green Algae (CCUBGA),
Indian Agricultural Research Institute (IARI), New Delhi-110 012, India
*Corresponding author, E-mail: prasannaiari@[Link]

Received on 26-12-2012 and Accepted on 26-02-2013

ABSTRACT: Nematicidal potential of ten terrestrial heterocystous and non heterocystous cyanobacteria were compared against
root-knot nematode, Meloidogyne incognita J2s. The extracts of all the ten unsonicated cyanobacteria caused an average immobility
of 4.8-22.9% and mortality of 1.5-5.6% which increased to 88.2-96.5% and 4.2-29.3% on sonication, respectively. The highest per cent
mean mortality was observed in S. nidulans (29.3) followed by P. tenue (26.8), M. tenera (25.8), T. nodosa (22.6), M. vaginatus (21.3), S.
bohmeri (17.5), N. punctiforme (6.5), A. variabilis (5.6), C. brevissima (5.1) and least in H. luteolus (4.2); with an increase in exposure time
from 24 to 72h. The immobility and mortality were significantly higher in sonicated extracts compared to unsonicated extracts.

Key words: cyanobacteria, extracts, heterocystous, non heterocystous, immobility, mortality. Meloidogyne incognita

Cyanobacteria are photosynthetic prokaryotic for their bionematicidal potential against root-knot
organisms that are cosmopolitan in habitat and constitute nematode, Meloidogyne incognita.
a substantial biomass in the soil. Heterocystous
cyanobacteria have been valued for their ability to fix MATERIALS AND METHODS
atmospheric nitrogen, bind and enrich soil and are
employed as biofertilizers (Venkataraman, 1972; Das, Culture of cyanobacteria (Blue green algae)
1991). Both heterocystous and non heterocystous forms
are reported to produce a large number of metabolites The heterocystous cyanobacteria viz; Hapalosiphon
which have been identified as toxins (microcystins, luteolus, Anabaena variabilis, Calothrix brevissima,
nodularins, neurotoxins), antibiotics, and protease Nostoc punctiforme, Tolypothrix nodosa, Scytonema
inhibitors. Majority of the known toxins are peptides, bohmeri and non heterocystous cyanobacteria viz;
alkaloids or non ribosomally synthesized molecules Synechococcus nidulans, Phormidium tenue,
(Namikoshi & Rinehart, 1996); some have been Microchaete tenera were obtained from Centre for
chemically characterized as neurotoxins causing inhibition Conservation and Utilisation of Blue Green Algae
of acetylcholine esterase and/or functions like (CCUBGA), I.A.R.I., New Delhi. The non heterocystous
acetylcholine (Carmichael, 1994). Few investigations cyanobacterium, Microcoleus vaginatus was obtained
have been carried out on cyanobacteria to evaluate their from Department of Botany, University of Allahabad,
bionematicidal potential (Dhanam et al., 1994; Gaur, Allahabad. The cyanobacteria were grown axenically in
1995; Khan et al., 1997; Sharma et al., 2007; Sharma & 1000 ml of in BG-11 (N-) medium for heterocystous
Gaur, 2008). A large number of cyanobacterial species species and BG-11(N+) medium (Stanier et al., 1971)
remain to be examined for their nematicidal effects. In for non heterocystous species in 3 l Haffkin flasks using
the present study, ten species of terrestrial heterocystous cotton plugs to allow aeration. The medium was sterilised
and non heterocystous cyanobacteria were evaluated at 121æ%C for 20 min at 103421.25 Pa pressure followed

34
 Evaluation of heterocystous and non heterocystous cyanobacterial species

by inoculation with 10 ml of fresh cyanobacterial culture immobility of nematodes was checked by probing the tail
and incubated in a culture room at 4000 lux with a 16 h:8 tip with a nematode pick and the immobile nematodes
h light/dark period at 28 ± 2æ%C for 14 days. The flasks were counted after 24, 48 and 72 h. Further, the extracts
were manually shaken each day to aerate the culture. of each exposure period were replaced with distilled
water, rinsing five times. After the final rinse, nematodes
Preparation of extracts of sonicated cyanobacteria were kept in distilled water for 24 h to observe if
nematode activity resumed. The numbers of active and
The sonication of microbial cells in suspension with inactive nematodes were counted to calculate the percent
high frequency sound waves results in their inactivation [Link] total chlorophyll and total soluble proteins
and disruption. One gram fresh weight of cyanobacterial of 10 cyanobacterial species were also estimated in 14-
biomass obtained from 14-day-old culture flasks was day-old cultures (Mckinney, 1941; Lowry et al., 1951;
concentrated by centrifugation (1008 g, 10 min) and was Herbert et al., 1971)
subjected to sonication in 2 ml Eppendorf tubes, using 1
ml sterile distilled water. Sonication was carried out Statistical analysis
intermittently (0.5 sec) using labsonic (B. Braun Biotech
International, Melsungen, Germany) at amplitude of 65 The data on immobility and mortality were analysed
Hz, four times for 7 min, at an interval of 7 min. The tubes by two factorial analysis of variance (ANOVA) and the
were maintained at low temperature by placing them in chlorophyll and total soluble protein by a single factor
compressed ice to avoid degeneration of the toxin during ANOVA (Gomez & Gomez, 1984). The data on percent
sonication. Sonicated cells were centrifuged again at immobility and mortality were arc sine transformed
1008 g for 10 min, the supernatants were collected and before analysis. The critical difference (CD) values at
the volume was made to 10 ml with sterile distilled water. 0.05 P were used to determine the significance of
The sonicated extract so obtained was used for nematicidal treatment mean differences.
assays.
RESULTS AND DISCUSSION
Preparation of extracts of unsonicated cyanobacteria
The unsonicated extracts of all the ten cyanobacteria
caused an average immobility in the range of 1.3-39.1%
The cyanobacterial cultures were centrifuged at within 72h. M. vaginatus, S. bohmeri and P. tenue
1008 g for 10 min. The supernatant was filtered through caused significantly higher percent immobility of 22.9,
Whatman No. 1 filter paper. The filtrate was used as a 21.5 and 19.4, respectively, compared to M. tenera
unsonicated extract for nematicidal assays (17.4) and T. nodosa (12.5). The percent immobility due
to S. nidulans (11.5) was at par with H. luteolus (9.7)
In vitro nematicidal activity and A. variabilis (9.5) but significantly higher than that
in N. punctiforme (5.3) or C. brevissima (4.8) compared
The cultures of all 10 cyanobacterial species were to 1.1 in the medium and 0.9 in the distilled water (Table
harvested at 14 days and tested for their nematicidal 1). The average per cent mortality in the unsonicated
efficacy with and without sonication against M. incognita extracts ranged from 1.5–5.6; the minimum for C.
J2s. Freshly hatched J2s of M. incognita, was obtained brevissima and maximum for S. bohmeri (Table 2). The
from eggmasses extracted from tomato plant pot cultures differences in per cent mortality were non-significant
maintained in the glasshouse of the Division of between all the treatments except for S. bohmeri which
Nematology, I.A.R.I., New Delhi. At least 100 nematodes showed significantly higher mortality than other extracts
in 0.5 ml sterile distilled water were poured into each well without sonication.
of the multiwell plate containing 1 ml cyanobacterial
extract, distilled water (control) and medium (control). Sonication significantly increased the toxic effect of
Three replications were maintained for each cyanobacteria on the nematodes. The sonicated extracts
cyanobacterial species and for each treatment. The of all the ten cyanobacteria caused an average immobility

35
Prasanna Holajjer et al.

Table 1. Effect of extracts from unsonicated heterocystous and non heterocystous cyanobacteria and exposure time on per cent
immobility of Meloidogyne incognita (J2)

Treatments % immobility of M. incognita (J2) after Mean

24h 48h 72h
Heterocystous cyanobactera
Anabaena variabilis 3.0(9.9) 8.3(16.5) 17.3(24.6) 9.5(17.0)
Calothrix brevissima 2.3(8.7) 3.2(10.1) 8.9(16.5) 4.8(11.8)
Hapalosiphon luteolus 3.7(11.1) 8.7(17.0) 16.7(24.0) 9.7(17.4)
Nostoc punctiforme 1.5(6.7) 7.0(15.2) 7.5(15.8) 5.3(12.6)
Scytonema bohmeri 20.5(26.7) 21.7(27.6) 22.4(28.1) 21.5(27.5)
Tolypothrix nodosa 5.5(13.3) 6.1(14.0) 25.8(30.2) 12.5(19.2)
Non heterocystous cyanobacteria
Microchaete tenera 1.3(6.3) 11.8(19.7) 39.1(38.6) 17.4(21.6)
Microcoleus vaginatus 11.2(19.4) 27.9(31.7) 29.7(32.9) 22.9(28.0)
Phormidium tenue 16.6(23.5) 18.0(25.1) 23.5(29.0) 19.4(25.9)
Synechococcus nidulans 4.0(11.4) 11.5(19.7) 18.9(25.4) 11.5(18.8)
Control
Medium 0.3(1.7) 1.4(5.5) 1.6(7.3) 1.1(4.9)
Water 1.1(4.9) 0.9(4.4) 0.7(3.9) 0.9(4.4)
Mean 5.9(12.0) 10.6(17.2) 17.7(23.0)
CD (0.05P)
(treatment) (3.6)
(time) (1.8)
(treatment x time) (6.2)
*Figures in parentheses are arc sine transformed values. Average of 3 replications.
Table 2. Effect of extracts from unsonicated heterocystous and non heterocystous cyanobacteria and exposure time on per cent
mortality of Meloidogyne incognita J2

Treatments % mortality of M. incognita (J2) after Mean

24h 48h 72h
Heterocystous cyanobactera
Anabaena variabilis 1.6(5.8) 2.8(9.4) 3.9(11.4) 2.8(8.9)
Calothrix brevissima 1.0(4.6) 1.4(5.4) 2.3(8.5) 1.5(6.2)
Hapalosiphon luteolus 0.5(2.4) 1.5(5.6) 2.7(9.1) 1.6(5.7)
Nostoc punctiforme 1.4(5.1) 1.4(6.8) 5.4(10.6) 2.7(7.6)
Scytonema bohmeri 3.7(11.1) 4.4(11.4) 8.5(16.9) 5.6(13.1)
Tolypothrix nodosa 1.9(6.4) 3.3(8.0) 3.9(10.6) 3.0(8.3)
Non heterocystous cyanobacteria
Microchaete tenera 2.1(8.0) 3.4(10.2) 5.9(13.9) 3.8(10.7)
Microcoleus vaginatus 2.0(6.7) 2.4(8.9) 3.8(10.0) 2.7(8.5)
Phormidium tenue 2.7(9.3) 3.9(11.1) 7.0(15.2) 4.5(11.9)
Synechococcus nidulans 1.8(7.5) 2.7(9.3) 4.8(12.5) 3.1(9.8)
Control
Medium 1.0(4.6) 0.0(0.0) 0.7(3.9) 0.6(2.9)
Water 0.0(0.0) 0.0(0.0) 0.4(2.1) 0.1(0.7)
Mean 1.6(6.0) 2.3(7.2) 4.1(10.4)
CD (0.05P)
(treatment) (3.9)
(time) (1.9)
(treatment x time) N.S
*Figures in parentheses are arc sine transformed values. Average of 3 replications.

36
 Evaluation of heterocystous and non heterocystous cyanobacterial species

in the range of 81.3-96.6% and occurred within 24 as sonication significantly enhanced the per cent mortality
exposed J2 were immobile within this time. After 72 h, of M. incognita juveniles (Dutta et al., 2007; Holajjer et
immobility increased in the range of 91.5-98.3% in al., 2012).
sonicated extracts compared to 1.6% in medium and
0.7% in water (Table 3). It appears from the observations Significant differences in mortality were also observed
that the cyanobacteria like M. vaginatus, S. bohmeri among the cyanobacterial species. The highest mean
and P. tenue which caused significant immobility in J2s mortality (29.3%) was observed in S. nidulans followed
of M. incognita even without sonication, released their by P. tenue (27.8%), M. tenera (25.8%), T. nodosa
toxic metabolites in the medium. Other cyanobacteria (22.6%), M. vaginatus (21.3%) and S. bohmeri (17.5%).
like, S. nidulans do not appear to release sufficient However, in some cyanobacteria, the per cent immobility
amount of their toxic metabolites in the medium as their was reversible or temporary. This was observed in A.
unsonicated extracts caused significantly lesser immobility variabilis, H. luteolus, C. brevissima and N. punctiforme
compared to M. vaginatus, S. bohmeri and P. tenue. It where the per cent immobility was 96.5, 95.7, 92.8 and
is only after the breakdown of the cells that the nematicidal 95.7 but the mortality values were 5.6, 4.2, 5.1 and 6.5%,
components are released in S. nidulans extract, as respectively.

Table 3. Effect of extracts from sonicated heterocystous and non heterocystous cyanobacteria and exposure time on per cent
immobility of Meloidogyne incognita (J2)

Treatments % immobility of M. incognita (J2) after Mean

24h 48h 72h

Heterocystous cyanobactera
Anabaena variabilis 96.6(80.0) 94.6(76.6) 98.3(82.9) 96.5(79.8)
Calothrix brevissima 90.8(72.6) 92.9(76.0) 94.6(77.7) 92.8(75.4)
Hapalosiphon luteolus 93.9(76.2) 95.2(77.6) 98.0(82.2) 95.7(78.7)
Nostoc punctiforme 94.8(77.1) 96.1(79.0) 96.1(78.8) 95.7(78.3)
Scytonema bohmeri 90.9(72.4) 91.4(73.4) 93.4(75.1) 91.9(73.6)
Tolypothrix nodosa 86.6(69.4) 87.9(69.8) 91.5(73.6) 88.7(70.9)
Non heterocystous cyanobacteria
Microchaete tenera 86.3(68.4) 91.8(73.7) 92.5(74.1) 90.2(72.1)
Microcoleus vaginatus 83.5(66.8) 86.3(69.9) 94.9(77.5) 88.2(71.4)
Phormidium tenue 81.3(64.9) 89.5(71.2) 94.6(76.9) 88.5(71.0)
Synechococcus nidulans 94.9(77.6) 93.1(76.1) 94.6(77.5) 94.2(77.0)
Control
Medium 0.3(1.7) 1.4(5.5) 1.6(7.3) 1.1(4.9)
Water 1.1(5.0) 0.9(4.4) 0.7(3.9) 0.9(4.4)
Mean 75.1(61.0) 76.8(62.8) 79.2(65.6)

CD (0.05P)
(treatment) (4.8)
(time) (2.4)
(treatment x time) N.S.
*Figures in parentheses are arc sine transformed values. Average of 3 replications.

37
Prasanna Holajjer et al.

Table 4. Effect of extracts from sonicated heterocystous and non heterocystous cyanobacteria and exposure time on per cent
mortality of Meloidogyne incognita (J2)

Treatments % mortality of M. incognita (J2) after Mean

24h 48h 72h

Heterocystous cyanobactera
Anabaena variabilis 3.3(10.3) 6.3(14.5) 7.1(15.4) 5.6(13.4)
Calothrix brevissima 3.6(10.8) 4.0(11.4) 7.7(15.6) 5.1(12.6)
Hapalosiphon luteolus 1.6(6.0) 2.0(8.1) 9.0(17.4) 4.2(10.1)
Nostoc punctiforme 4.9(12.5) 5.5(13.6) 9.1(17.0) 6.5(14.3)
Scytonema bohmeri 16.2(23.7) 17.0(24.3) 19.1(25.7) 17.5(24.5)
Tolypothrix nodosa 20.5(26.8) 21.6(27.7) 24.7(29.6) 22.6(28.0)
Non heterocystous cyanobacteria
Microchaete tenera 22.0(27.8) 26.4(30.9) 29.1(32.6) 25.8(30.4)
Microcoleus vaginatus 13.0(21.0) 17.8(24.8) 33.3(35.1) 21.3(27.0)
Phormidium tenue 24.1(29.2) 26.3(30.7) 30.0(33.1) 26.8(31.0)
Synechococcus nidulans 13.3(21.1) 32.2(34.5) 42.4(40.6) 29.3(32.0)
Control
Medium 1.0(4.6) 0.0(0.0) 0.7(3.9) 0.6(2.8)
Water 0.0(0.0) 0.0(0.0) 0.4(2.1) 0.1(0.7)
Mean 10.6(16.4) 13.3(18.4) 17.7(22.3)

CD (0.05P)
(treatment) (3.1)
(time) (1.6)
(treatment x time) (5.4)
*Figures in parentheses are arc sine transformed values. Average of 3 replications.

An attempt was made to compare the mortality of Table 5. Chlorophyll and total soluble protein content of
juveniles with the protein and chlorophyll content of the heterocystous and non heterocystous cyanobacteria
cyanobacteria. The data on total chlorophyll (µg/g fresh Treatment Chlorophyll Total soluble
weight) and total soluble protein (mg/g fresh weight) are (µg/g fresh protein(mg/g
given in Table 5. The highest chlorophyll content was weight) fresh weight)
observed in H. luteolus (273.6 µg/g), which was at par Heterocystous cyanobacteria
with that in S. nidulans (268.8 µg/g) at CD(0.05P)=21.5.
Anabaena variabilis 214.2 41.2
The highest protein content (48.8 mg/g) was observed in Calothrix brevissima 171.7 16.1
N. punctiforme, which was at par with that in H. Hapalosiphon luteolus 273.6 47.2
luteolus (47.2 mg/g), which were significantly higher Nostoc punctiforme 202.5 48.8
than that in S. nidulans (18.4mg/g). Thus, both chlorophyll Scytonema bohmeri 128.7 38.2
and protein content could not be related to the toxic Tolypothrix nodosa 231.8 7.2
effects of the cyanobacteria. . Non heterocystous cyanobacteria
Microchaete tenera 31.6 20.1
The nematicidal culture filtrates or cell extracts from Microcoleus vaginatus 199.5 30.8
cyanobacteria can be formulated using suitable carriers Phormidium tenue 69.7 18.5
for seed or soil treatment as protectants against plant Synechococcus nidulans 268.8 18.4
parasitic nematodes. The nematicidal effect of S. CD (0.05P) 21.5 5.2

38
 Evaluation of heterocystous and non heterocystous cyanobacterial species

nidulans using formulations has been investigated in Robbins (ed.) Methods in Microbiology. Vol. VB.
microplot experiments against M. incognita, infecting Academic Press, New York, pp. 209.
brinjal, wherein significant reduction in nematode Holajjer, P., Kamra, A., Gaur, H.S. & Dhar, D.W. (2010).
multiplication and root galling was observed (Holajjer et Bioefficacy and shelf life of carrier based formulations
al., 2010). of cyanobacterium, Synechococcus nidulans against
Meloidogyne incognita infecting brinjal. International
ACKNOWLEDGMENTS Journal of Nematology 20: 162-168.

The authors are grateful to the Centre for Holajjer, P., Kamra, A., Gaur, H.S. & Dhar, D.W. (2012). In
vitro nematicidal activity of a terrestrial cyanobacterium,
Conservation and Utilisation of Blue Green Algae Synechococcus nidulans towards plant-parasitic
(CCUBGA), I.A.R.I., New Delhi and Department of nematodes. Nematology 14: 85-92.
Botany, University of Allahabad, Allahabad for providing
the cyanobacterial cultures. Khan, Z., Jairajpuri, M.S. & Khan, M.W. (1997). Effect of
culture filtrate to a blue-green alga, Microcoleus
vaginatus on the mortality and hatching of M. incognita.
REFERENCES International Journal of Nematology 7: 100-102.

Carmichael, W.W. (1994). The toxins of cyanobacteria. Khan, Z., Jairajpuri, M.S. & Khan, M.W. (1999). Efficacy of
Scientific American 270: 78-86. seed soaking with culture filtrate of a blue green alga,
Microcoleus vaginatus for the control of root-knot
Chandel, S.T. (2009). Nematicidal activity of the nematode, Meloidogyne incognita on okra. Indian
Cyanobacterium, Aulosira fertilissima on the hatch of Journal of Nematology 29: 82-84.
Meloidogyne triticoryzae and Meloidogyne incognita.
Archives of Phytopathology and Plant Protection 42: Lowry, O.H., Rosebrough, N.J., Farr, A.L. & Randall, R.J.
32-38. (1951). Protein measurement with the Folin phenol
reagent. Journal of Biological Chemistry 193: 265-275.
Das, H.K. (1991). Biological nitrogen-fixation in the context of
Indian agriculture. Current Science 60: 551-55. Mckinney, G. (1941). Absorption of light by chlorophyll
solutions. Journal of Biological Chemistry 140: 315-
Dhanam, M., Kumar, A.C. & Sowjanya. (1994). Microcoleus 322.
vaginatus (Oscillatoriaceae), a blue-green alga (or
Cyanobacterium) parasitizing plant and soil nematodes. Namikoshi, M. & Rinehart, K.L. (1996). Bioactive compounds
Indian Journal of Nematology 24: 125-132. produced by cyanobacteria. Journal of Industrial
Microbiology 17: 373-384.
Dutta, T.K., Kamra, A., Gupta, A.L., Gaur, H.S. & Dhandapani, Sharma, H.K. & Gaur, H.S. (2008). Hatch inhibition of
A. (2007). Toxicity of cyanobacterium, Synechochoccus Meloidogyne incognita by aqueous extracts and
(Anacystis) nidulans extracts on plant-parasitic exudates of five species of cyanobacteria. Nematologia
nematodes. International Journal of Nematology 17: Mediterranea 36: 99-106.
137-143.
Sharma, H.K., Gaur, H.S., Singh, D.V. & Dhar, D.W. (2007).
Gaur, H.S. (1995). Studies on the hatch inhibition and Bionematicidal potential of ten species of cyanobacteria
nematicidal action of exudates and extracts of against root-knot nematode, Meloidogyne incognita.
cyanobacteria and a green alga. Annals of Plant International Journal of Nematology 17: 35-40.
Protection Sciences 32: 158-163.
Stanier, R.Y., Kinisawa, R., Mandel, M. & Choen-Bazire, G.
Gomez, K.A. & Gomez, A.A. (1984). Statistical procedure for (1971). Purification and properties of unicellular blue
agricultural research (2nd Eds). John Wiley and Sons, green algae (Order: Chroococcales). Bacteriological
New York. Reviews 35: 171-205.

Herbert, D., Phipps, P.J. & Strange, R.E. (1971). Chemical Venkataraman, G.S. (1972). Algal biofertilizeres and rice
analysis of microbial cells. In: J. R. Norris and D. W. cultivation. Today and Tomorrow Printers, Haryana,
India.

39
B.S. Sunanda and A.U. Siddiqui
Vol. 43, No. 1, pp. 40-42 Indian Journal of Nematology June, 2013

In vitro Production of Steinernema carpocapsae in Different Artificial Media

B.S. SUNANDA AND A.U. SIDDIQUI

Department of Nematology, Rajasthan College of Agriculture, Maharana Pratap University of Agriculture & Technology,
Udaipur-313 001, India

Received on 06-01-2013 and Accepted on 01-03-2013

ABSTRACT: The successful implementation of entomopathogenic nematodes (EPN) in the families Steinernematidae and
Heterorhabditidae against soil inhabiting insects is due to their ease of culture. Mass multiplication of S. carpocapsae was studied
(in vitro) on different artificial media i.e., Wout’s, Modified egg yolk and Egg yolk media. Out of these highest (25.88) lakhs IJs were
obtained from Wout’s media followed by modified egg yolk (20.78) lakhs and Egg yolk (0.91) lakhs media. It was observed that S.
carpocapsae multiplied on both animals as well as plant protein based media.

Key words: Entomopathogenic nematodes, Steinernema carpocapsae, In vitro, artificial media, mass production.

Rhabditid nematodes of the families sponges (1.5 cm3). The flasks were filled with foam
Heterorhabditidae and Steinernematidae are potential chips medium mixture (1.5 g of foam chips: 8-9 g of
biocontrol agents of various insect pests, (Poinar, 1979; medium, w/w) and plugged tightly with cotton. The flasks
Klein, 1990; Kaya et al., 1993). They provide some of the were autoclaved for 20 min. at 121°C and allowed
most promising new alternatives to chemical insecticides cooling at room temperature before inoculation with
and are non-polluting, non toxic and environmentally infective juveniles (IJs) fresh IJs extracted from the
friendly with comparative safety to non-target and infected insect cadavers were used.
beneficial organisms. Although these nematodes are
easily produced in vitro on various complex semi-solid The nematodes were inoculated aseptically @ 1000
organic media. The successful implementation of IJs/flask. Care was taken not to shake the flasks after
entomopathogenic nematodes (EPN) in the families the inoculation of nematodes. The sealed flasks were
Steinernematidae and Heterorhabditidae against soil incubated at 28°C for 30 days. Colonies of the nematodes
inhabiting insects is due to their ease of culture (Georgis, were observed on the walls of the flasks after 20 days
1990). The associated bacteria convert a wide range of post inoculation. The harvesting of the nematodes was
substrates into media suitable for nematode development done after 30 days. The nematode yield from each
and the nematode-bacterium complex is culturable on a treatment medium harvested was expressed in terms of
variety of non-restrictable artificial media (Bedding, number of IJs/flask.
1981; Friedman, 1990). Mass production at low cost is a
pre requisite for the use of EPN. In vitro methods using Wout’s medium: Nutrient-broth 0.88 g; Yeast-extract
artificial media come handy for mass production of IJs in 0.32 g; Groundnut-oil 10.40 g; Soy flour 14.40 g; Distilled-
large numbers. Protein and fatty acids play a vital role in water 60 ml.
the progeny production of EPN.
Egg yolk medium: Solid egg yolk 7.00 g; Yeast-extract
MATERIALS AND METHODS 2.00 g; Sodium chloride 0.80 g; Groundnut oil 15.00 g;
Distilled-water 60 ml.
The ingredients were mixed together according to
different media composition with polyether polyurethane

40
 In vitro production of Steinernema carpocapsae

Modified egg yolk medium: Egg yolk 7.00 g; Soy flour maximum 64.6 × 105 and 48.9 × 105 IJs/250 ml flask.
20.00 g; Yeast-extract 2.00 g; Sodium chloride 0.80 g; Similar studies were made on in vitro mass multiplication
Groundnut oil 15.00 g; Distilled-water 60 ml. The and estimation of cost production of Steinernema seemae
observation was taken after 30 days of inoculation. Ali et al. (2008) which cost Rs. 2.20 to get 60 × 105 IJs.
Vyas et al (2010) worked out in vitro production cost of
RESULTS AND DISCUSSION S. glaseri for 1 million IJs of S. glaseri for Rs. 0.25 in
egg yolk and Rs. 0.72 in dog food biscuit media. As
The mass multiplication (in vitro) of S. carpocapsae for as economics of mass production of infective juveniles
was done on different artificial media. The infective is concerned out of these artificial media were taken, out
juveniles of S. carpocapsae were inoculated @ 1000 of which Wout’s media was found more economical than
IJs/ 250ml flask. It was observed that IJs multiplied both modified egg yolk and egg yolk media in respect to cost
on plant and animal protein media. Out of these media involved and production of IJs/250 ml flask. The Wout’s
Wout’s media was found better as compared to other media was synthesized using locally available ingredients
media. The maximum number (in lakh) of S. carpocapsae such as nutrient broth 0.88 g, yeast extract 0.32 g,
obtained in Wout’s media (25.88) lakhs followed by groudnut oil 10.40 g, soyflour 14.40 g and distilled water
Modified egg yolk (20.83) lakhs and Egg yolk media 60 ml which has costed Rs. 12.20 to get 25.88 lakh IJs of
(0.91) lakhs. Similar studies in this regard were made by S. carpocapsae as against modified egg yolk and egg
Hussaini et al. (2000) who reported maximum yield yolk media costed 16.20 and Rs. 18.50 to obtain 20.78
(30.58 × 105) of S. carpocapsae in Wout’s media as and 0.91 lakh IJs respectively.
compared to other media. Yadav (2006) also studied the
mass multiplication of S. carpocapsae on artificial Economics of in vitro mass production of S.
media, and recorded maximum (9.37 × 105) yield from carpocapsae on different artificial media
Wout’s media. However, Vyas et al. (2001) tested 21
animals and plant protein based media for the mass Media Composition production cost/
production of S. carpocapsae and recorded poor yield 250 ml flask(Rupees)
of Steinernema sp. in plant protein as compared to
Wout’s Rs. 12.20 (25.88 lakh IJs)
animal protein based media. Hussaini (2007) tested
Wout’s medium for in vitro mass multiplication of Egg yolk Rs. 16.80 (20.78 lakh IJs)
indigenous isolate of S. carpocapsae was obtained in Modified egg yolk Rs. 14.20 (0.91 lakh IJs)
medium supplemented with better Soya and Sunflower

Table 1: Yield of Steinernema carpocapsae obtained on different artificial media (30 DAI)

Treatments Yield of S. carpocapsae (in lakhs)

2010-11 2011-12 Pooled Mean

T1 = Wout’s medium (1000 IJs/250 ml flask) 25.75 26.00 25.88

T2 = Modified egg yolk medium (1000 IJs/250 ml flask) 21.55 20.80 20.78
T3 = Egg yolk medium (1000 IJs/250 ml flask) 0.81 1.00 0.91
SEm± 0.49 0.50 0.35
CD at 5% 1.56 1.60 1.04

(Treatments of 4 replications)

41
B.S. Sunanda and A.U. Siddiqui

REFERENCES Hussaini, S.S., Nagesh, M., Rajeshwari, R. & Manzoor, H.

(2007). Effect of Protein and Lipid Sources in the Wout’s
Ali, S.S., Pervez, R. Hussain, M.A. & Ahmad, R. (2008). Medium on the Yield and Pathogenicity of Steinernema
Susceptibility of three lepidopteran pest to five carpocapsae and S. tami, Indian Journal of Plant
entomopathogenic nematodes and in vivo mass Protection 35: 93-96.
production of these nematodes. Archieves of
Phytopathology and Plant Protection 39: 17-22. Kaya, H.K., Burlando, T.M. & Thurston, G.S. (1993). Two
entomopathogenic nematode species with different
Bedding, R.A. (1981). Low cost in-vitro mass production of search strategies for insect suppression. Environmental
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114 Klein, M.G. (1990). Efficacy against soil inhabiting insect
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Friedman, M.J. (1990). Commercial production and Entomopathogenic nematodes in biological control.
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CRC Press: Boca Raton, Florida, U.S.A., 153-172. Poinar, G.O. Jr. (1979). Nematodes for biological control of
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Georgis, R. (1990). Formulation and application technology,
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Raton, Florida. U.S.A. 173-191. to Maharana Pratap University of Agriculture and
Technology, Udaipur, Rajasthan.
Hussaini, S.S., Kavita Satya, J. & Hussaini, M.A. (2000).
Mass production of a native Steinernema sp. (SSL2) Vyas, R.V. (2001). In vitro mass production of native
PDBC EN 13.21 (Nematode: Steinernematidae) on Steinernema sp. Annals of Plant Protection Sciences 9:
different artificial media. Indian Journal of Plant 77-80.
Protection 28: 94-96.

42
 Methanol neem extracts on mortality of Meloidogyne incognita
Vol. 43, No. 1, pp. 43-46 Indian Journal of Nematology June, 2013

Effect of Methanol Neem Extracts on Mortality of Meloidogyne incognita

Juvenile
NARESH NAYAK* AND J.L. SHARMA

P.G. Department of Botany, Govt. College, Kota, (Raj.), India

*Corresponding author, E-mail: nayak.nema2009@[Link]

Received on 10-01-2013 and Accepted on 20-03-2013

ABSTRACT: Root-knot nematode Meloidogyne incognita is regarded as one of the obstacles in the agriculturel productivity. Natural
products are used as safer alternative to control root knot nematode. An experiment was carried out in vitro to test the neem plant
parts methanol extract. Different concentration of shade-dried leaves, bark and seeds of neem were tested for mortality of Meloidogyne
incognita juveniles. Increase in concentration and exposure period resulted in increased mortality rate of Juveniles. Methanol neem
seed (5 ml concentration) proved to be the most effective among used plant parts viz. bark, leaves, and seed. Neem bark was the least
effective among the three.

Key words: M. incognita, Methanol, Plant extract, Mortality

The root-knot nematode, Meloidogyne incognita overnight, powder of plant parts were prepared by using
is a serious problem to the cultivation of agricultural crop a mixer or blender. Five gm powder of neem bark, neem
through out the world. It causes severe damage and yield leaves and neem seeds were soaked in 50 ml methanol
loss to a wide group of vegetable crop in all parts of India for 7-9 days for evapouratation after that it was washed
(Khan & Khan, 1996). Almost all the chemical with distilled water and absolute organic solvents. Extract
nematicides are expensive and create environmental were filtered through four ply muslin cloth and then
problems. Modern agriculture moves to adopt more passed through Whatman filter paper no.1 filtered extract
environment friendly practices. There is increasing were make up 50 ml with required amount of distilled
interest in the use of biopesticides that are pest specific water and then centrifuged at 4000 rpm for 10 minutes
and non toxic to human and beneficial organisms. Extract then again filtered through Whatman filter paper no.1.
of plant product contain nematicidal compound. Thousand The extract so obtained was put in conical flasks and
of plants possessing insecticidal properties are known steam sterilized in an autoclave. The plant part extract
today (Banerji et al. 1985). Use of neem and neem viz. neem bark, neem leaves and neem seed designated
product has been advocated for the management of root- as stock solution for preparing different dilution. The
knot nematode by many workers (Dekha & Rehman, sterilized distilled water served as control.
1998). Therefore the present investigation was known
about the nematicidal effect of neem plant parts as Effect of methanol extracts on juvenile mortality of
methanol extract on mortality of M. incognita juveniles. M. incognita: Seven dilutions viz. (5 ml, 2.5ml, 2 ml, 1.5
ml, 1 ml, 0.50 ml, 0.25 ml) were prepared from stock
MATERIALS AND METHODS solution. Five ml of each plant parts methanol extracts
were prepared by adding the required amount of distilled
Preparation of Extracts: The Neem (Azadirachta water were taken in sterile beaker of 50 ml capacity.
indica) plant parts viz. neem bark, neem leaves, neem Two hundred freshly hatched second stage juveniles J2s
seeds were shade dried and then oven dry at 60oC for of M. incognita were transferred in beaker. Equal

43
Naresh Nayak and J.L. Sharma

number of J2s was also transferred to separate beaker concentration had only 54% mortality after 120 h of
containing sterilized water to serve as control. Three exposure period.
replicated were taken completely randomized design
(CRD). Observations of both live and dead nematodes Cumulative percent mortality of M. incognita was
were made at 24, 48, 72, 96 and 120 h after inoculation. maximum in neem seed extract and minimum in neem
bark extract which could be due to the fact that Neem
RESULTS AND DISCUSSION bark does not possess sufficient active ingredient which
are nematicidal. Neem leaves and Neem seed having
Result present in table (1, 2 & 3) revealed the more concentrated nematicidal/nematostatic properties.
maximum mortality was recorded in 5ml concentration The present investigation in adjustable conformity with
which increased with increase in methanol extract con. the finding of Vijayalaxmi et al. (1979) found leaf extract
more number or percentage of juveniles mortality with of A. indica caused increased Mortality of M. incognita
increase in exposure period too. Minimum mortality per juvenile. These results also agree with the Joymatidevi
cent was observed in neem bark methanol extract (Table (2010) on effect of ten different medicinal plants methanol
1) there was 81 percent mortality at its highest 5ml con. extract against second stage juveniles of M. incognita.
and 24 h exposure period. Only 24 percent mortality Haseeb et al. (2007) reported that A. indica seeds
recorded at its lowest concentration (0.25ml) after 120 powder were found effective against M. incognita.
hrs of exposure period. Highest concentration of neem Joymatidevi (2008) tested methanol extracts in laboratory
leaf extract (5ml) revealed that cent percent mortality condition and found A. indica and Melia azaderach
after 48 h of exposure period and 41.66 percent mortality show highest larval mortality. Neem producte (Neem
recorded at it lowest concentration (0.25ml) after 120 h Kernel, Neem leaf, Neem bark extract) are known to
of exposure period. Neem seed methanol extract on the posses nematicidal activity against nematode population
other hand was found to be the most effective in (Zaki & Bhatti, 1989, Derkar et al., 1990). The present
juvenile’s mortality (Table 3), the highest concentration study also revealed that the mortality rate was increased
(5ml) were found to be most effective caused cent with increased expose period and concentration as
percent mortality from 24 h exposure and its lowest reported by Siddiqui & Alam (1988). Singh & Sitaramaiah

Table 1. Effect of Neem Bark Extract (Methanol) on juveniles Mortality of Meloidogyne incognita at different time intervals
(Observations are mean of three replicates)

Extract Con. % Mortality at different exposure period

(ml) 24 h 48 h 72 h 96 h 120 h

0.25 4.000 12.333 18.000 22.000 24.000

0.50 5.667 17.333 21.000 28.000 35.667
1.0 13.667 19.667 39.667 48.667 55.333
1.5 21.000 36.000 53.000 61.333 69.333
2.0 32.333 41.667 60.333 64.667 75.333
2.5 27.000 55.333 66.333 78.000 81.667
5.0 81.000 87.333 91.000 93.000 94.667
Control 0.000 0.000 0.000 0.000 0.000
SEM ± 5.923 1.258 2.309 1.080 1.354
C.D. (P=0.05) 13.659 2.902 5.325 2.491 3.122

44
 Methanol neem extracts on mortality of Meloidogyne incognita

Table 2. Effect of Neem Leaf Extract (Methanol) on juveniles Mortality of Meloidogyne incognita at different time intervals
(Observations are mean of three replicates)

Extract Con. % Mortality at different exposure period

(ml) 24 h 48 h 72 h 96 h 120 h

0.25 3.000 15.000 24.667 32.000 41.667

0.50 8.000 21.667 30.667 39.000 52.000
1.0 14.333 31.667 43.333 52.667 64.000
1.5 27.000 44.667 55.667 63.333 73.333
2.0 49.333 57.000 73.000 84.000 90.667
2.5 75.333 85.667 92.667 99.000 100.000
5.0 98.333 96.667 100.000 100.000 100.000
Control 0.000 0.000 0.000 0.000 0.000
SEM ± 1.236 2.055 1.724 1.280 1.633
C.D. (P=0.05) 2.850 4.738 3.976 2.952 3.766

Table 3. Effect of Neem Seed Extract (Methanol) on juveniles Mortality of Meloidogyne incognita at different time intervals
(Observations are mean of three replicates)

Extract Con. % Mortality at different exposure period

(ml) 24 h 48 h 72 h 96 h 120 h

0.25 9.000 20.333 38.000 46.667 54.000

0.50 18.000 30.667 47.333 52.000 69.667
1.0 30.333 42.333 55.000 66.000 78.333
1.5 43.000 58.667 72.000 78.333 87.667
2.0 74.000 81.667 86.333 92.667 100.000
2.5 83.333 97.333 100.000 100.000 100.000
5.0 100.000 100.000 100.000 100.000 100.000
Control 0.000 0.000 0.000 0.000 0.000
SEM ± 1.143 1.528 1.247 1.354 1.190
C.D. (P=0.05) 2.635 3.522 2.876 3.122 2.745

(1973) and Khan [Link]. (1974) reported that the responsible for mortality of M. incognita juveniles. It is
nematostatic and nematicidal properties of Neem therefore concluded that the incorporation of plant product
products might be due to azadirachtin as in active like neem could provide a safer, suitable and cheaper
principles and toxic chemicals present in them, which alternative to root-knot nematode M. incognita.
could have nematicidal/nematostatic effect. It was

45
Naresh Nayak and J.L. Sharma

ACKNOWLEDGEMENT 37: 19-22.

I am highly grateful to Head, PG Department of Joymatidevi (2008). Evaluation of chloroform methanol extracts
of medicinal plant on egg hatching and larval mortality
Botany Govt. College, College, Kota for providing
of Meloidogyne incognita. Indian Journal of
necessary facilities. Nematology 38: 168-171.

REFERENCES Khan, A.A. & Khan, M.W. (1996). Distribution of root-knot

nematode species and races infesting vegetable crop in
Banerji, R., Mishra, G. & Nigam, S.K. (1985). Role of eastern Uttar pardesh. Indian Journal of Nematology
indigenous plant material in pest control. Pesticides 19: 26: 238-244.
32-38.
Siddiqui, M.A. & Alam, M.M. (1988). Toxicity of different plant
Deka, B.C. & Rahman, M.F. (1998). Management of parts of Tagetes lucida to plant parasitic nematodes.
Meloidogyne incognita infecting okra by using neem Indian Journal of Nematology 18: 181-185.
part & products. Journal of Agriculture Science &
Society North East India. 11, 124-126. Vijayalakshmi, K., Mishra, S.D. & Prasad, S.K. (1979).
Nematicidal properties of some indigenous plant materials
Derker, K.S., Mhase, N.L. & Shelka, S.S. (1990) effect of against second stage juveniles of Meloidogyne
placement of non edible oilseed cake on the control of incognita (Kofoid and white, 1919) chitwood, 1949.
root-knot nematode. Indian Nematology Network Indian Journal of Engineering 41: 326-333.
Newsletter 7: 5-70.
Zaki, F.A. & Bhatti, D.S. (1989). Phytotherapeutic effect of
Haseeb, A., Amen, A. & Sharma, A. (2007). Management of some plant leaves on Meloidogyne javanica infecting
Meloidogyne incognita. Fusariun oxysporum Disease tomato plants. Nematologia Mediterranea 17:71-72.
complex of Pisum sativum. Indian Journal of Nematology

46
 Check-list of globally known species of Tylenchorhynchus
Vol. 43, No. 1, pp. 47-60 Indian Journal of Nematology June, 2013

A Check-List of Globally Known Species of Tylenchorhynchus Cobb, 1913

along with Compendium of the Indian Species
SUDERSHAN GANGULY*, M. LAL AND K.S. RATHOUR

Division of Nematology, Indian Agricultural Research Institute, New Delhi-110 012, India
*Corresponding author, E-mail: sg_nema@[Link]

Received on 22-02-2013 and Accepted on 30-03-2013

ABSTRACT: Stunt nematodes of the genus Tylenchorhynchus are widely distributed in association with almost all the field and
horticultural crops, and about 158 species are known globally, of which 60 species have been described from India. This paper provides
a checklist of all the known species of Tylenchorhynchus, along with compendium of the nominal Indian species and other recorded
species of genus from India.

Keywords: Checklist, compendium, crop host, locality, morphometrics, Species, Stunt nematodes, synonyms, taxonomy, India.

The soil dwelling stunt nematodes, earlier sometimes areolated. Mahajan (1988) gave a diagnostic
characterized under one genus Tylenchorhynchus are compendium to species of Tylenchorhynchus and
now split into eight more genera namely, Bitylenchus, included 89 valid species in the genus. Esser (1991) listed
Dolichorhynchus, Quinisulcius, Merlinius, 257 nominal species in his checklist of the genus. Brzeski
Amplimerlinius, Triversus and Trilineelus. Most of the and Dolinski (1998) compiled a compendium containing
information, however, on their association with diseased 177 species with 2 to 5 lines in the lateral field. The
conditions of plants is under the former complex genus, taxonomy of stunt nematodes has been advanced through
Tylenchorhynchus. Members of these genera possess scanning electron microscopy (Fortuner and Luc, 1987;
similar anatomy and may be easily mistaken for one Powers, 1983; Powers et al., 1983; Siddiqi, 1986).
another. Handoo (2000) prepared a key and a compendium
containing morphometric and related details to facilitate
The cosmopolitan genus Tylenchorhynchus was easy identification of 111 valid species of
established by Cobb (1913) for T. cylindricus found in Tylenchorhynchus. As a result of the inevitable
southern California. Allen (1955) enhanced taxonomic taxonomic changes, development of a dichotomous key
criteria for Tylenchorhynchus. Tarjan (1973) gave a to Tylenchorhynchus spp. has become increasingly
synopsis, key, and diagnostic data of the genera and difficult. Many species previously included in
species in the Subfamily Tylenchorhynchinae. The Tylenchorhynchus have been placed in newer genera,
history of the genus was discussed by Hooper (1978), and numerous species within related genera have been
Golden et al., (1987), and Anderson and Potter (1991). either shifted to Tylenchorhynchus or synonymized
Siddiqi (1986 & 2000) recognized 71 and 104 total with other species. The most important character used in
species, respectively as being in Tylenchorhynchus. distinguishing these genera is the number of lateral lines
Fortuner and Luc (1987), in their reappraisal of Tylenchina, or incisures, which can range from three to six.
included Tylenchorhynchus under the family
Telotylenchidae, subfamily Telotylenchinae, and In the present study, we compiled a check list of 158
recognized 129 valid species, defining Tylenchorhynchus globally known nominal species of Tylenchorhynchus,
as having 2 to 5 lines in the lateral field, which is (Fig. 1), listed 47 species which are either transferred to

47
Sudershan Ganguly et al.

other genera or synonymized with other genera/ species Neodolichorhynchus Jairajpuri & Hunt, 1984
(Siddiqi, 2000) and other species reported from India
(Fig.1 & Table 1). We also present a check-list and Paratrophurus Arias, 1970
compendium of 60 nominal Indian species of
Tylenchorhynchus containing morphometric and related Quinisulcius Siddiqi, 1971
details to facilitate easy identification (Table 2 & 3).
Sauertylenchus Sher, 1974

Telotylenchoides Siddiqi, 1971

Species described

Trichotylenchus Whitehead, 1960

Trophurus Loof, 1956

Tylenchorhynchus Cobb, 1913

Uliginotylenchus Siddiqi, 1971

Year
Genus Tylenchorhynchus Cobb, 1913
Fig. 1. Tylenchorhynchus species described in different
Diagnosis: Telotylenchinae. About 1 mm or less long.
decades along with the cumulative number of species
Cuticle prominently annulated may be marked by
longitudinal striae, longitudinal ridges or lamellae outside
Subfamily Telotylenchinae Siddiqi, 1960 lateral fields absent; not abnormally thickened on tail.
Lateral field with three or four incisures, generally not
Diagnosis: Lateral fields each with three to five incisors. areolated behind oesophageal region. Cephalic region
Amphidial aperture pore-like, labial, indistinct. Derids offset from body (type genus) or continuous, annulated,
absent. Stylet small to moderately long ( about 11-40µm or rarely smooth, without longitudinal indentations of
), with conus about as long as shaft and distinct basal annules; labial disc indistinct, in SEM en face squarish,
knobs. Median oesophageal bulb well developed; flattened, fused with first lip annule; framework light to
Oesophageal glands enclosed in a basal bulb or dorsal moderately sclerotized. Stylet well developed, generally
gland enlarged, formimg a long lobe extending over 15-25 µm long, knobs prominent. Median bulb round or
intestine. Vulva slit like, lacking epiptygma. Vagina not oval, with distinct refractive thickenings, usually marked
sclerotized. Ovaries paired except Trophurus. Female off from pre-corpus and isthmus. Basal bulb offset from
tail Conoid, subcylindroid or subclevate, rarely spicate, intestine, or base with slightly extending over intestine.
between two and six anal body widths long. Male tail Cardia prominent. Vulva median; Ovaries paired,
conical, about as long as that of female. Bursa simple, outstretched. Female tail usually conoid with blunt tip as
well developed, enveloping tail. Hypoptygma absent. in type species, but may be subcylindroid, cylindroid or
Spicules arcuate, with large distal flanges and pointed or subclavate; terminus smooth, rarely striated or narrowing
indented tip. Gubernaculum rod-like, usually protrisible, to a point. Males generally present. Tail enveloped by a
distally boat shaped in lateral view, with or without crust. large simple bursa. Spicules distally flanged.
Type genus: Telotylenchus Siddiqi, 1960 Gubernaculum well developed, about half as long as
Other genera: Bitylenchus Filipjev, 1934 spicule, generally rod-like and with a velum, protrusible.

Histotylenchus Siddiqi, 1971

48
 Check-list of globally known species of Tylenchorhynchus

Table 1. A check-list of globally known species of Tylenchorhynchus Cobb, 1913.

[Link] Species Author Year Remarks

1. aduncus de Guiran 1967
2. aerolatus Tobar Jime´nez 1970
3. agri Ferris 1963
4. alami Shaw & Khan 1996
5. allii Khurma & Mahajan 1987 (syn. of Bitylenchus quaidi)
6. amgi Kumar 1981
7. ancorastyletus Ivanova 1983
8. annulatus a (Cassidy, 1930) Golden 1971
9. antarcticus Wouts & Sher 1981
10. ascicaudatus Chang 1991
11. aspericutis Knobloch 1975
12. badliensis Saha & Khan 1982
13. bambusi Singh, Lal, Rathour & Ganguly 2010
14. berberidis Sethi & Swarup 1968 (syn. with Nagelus hexagramus)
15. bicaudatus Khakimov 1973
16. bohrrensis Gupta & Uma 1980
17. brassicae Siddiqi 1961
18. brevilineatus a Williams 1960 (Transferred to Bitylenchus)
19. bryobius Sturhan 1966 (Transferred to Bitylenchus)
20. cacti Chawla, Bhamburkar, Khan & Prasad 1968 (Transferred to Quinisulcius)
21. canalis Thorne & Malek 1968 (Transferred to Bitylenchus)
22. chirchikensis Mavlyanov 1978
23. chonai Sethi & Swarup 1968
24. cicerus Kakat, Khan & Siddiqi 1995
a
25. clarus Allen 1955
26. clathrocutis (Lewis & Golden) Fortuner & Luc 1987
27. clavicaudatus Seinhorst 1963 (Transferred to Bitylenchus)
28. clavus Khan 1990
29. claytoni Steiner 1937
30. coffeae Siddiqi & Basir 1959
31. contractus Loof 1964
32. crassicaudatus Williams 1960
33. cristatus Ivanova 1983 (Transferred to
Neodolichorhynchus)
34. crotoni Pathak & Siddiqi 1997
35. cuticaudatus Ray & Das 1983 (Transferred to Bitylenchus
36. cylindricus Cobb 1913
37. cynodoni Kumar 1981
38. dactylurus Das 1960
39. delhiensis Chawla, Bhamburkar, Khan & Prasad 1968

49
Sudershan Ganguly et al.

[Link] Species Author Year Remarks

40. depressus Jairajpuri 1982 (Transferred to Bitylenchus
41. dewaeli Kleynhans 1992 (Transferred to Telotylenchus)
42. digittatus Das 1960
43. dispersus Siddiqi & Sharma 1995
44. divittatus Siddiqi 1961
45. dubius a (Butschli, 1873) Filipjev 1936 (Transferred to Bitylenchus)
46. ebriensis Seinhorst 1963
47. elamini Elbadri et al., 2010
48. elegans Siddiqi 1961
49. eremicolus Allen 1955
50. erevanicus Karapetjan 1979
51. eroshenkoi Siddiqi 1986
52. estherae Kleynhans 1992 (Transferred to
Neodolichorhynchus)
53. ewingi a Hooper 1959
54. fugianensis Chang 1990
55. georgiensis Eliashvili 1971
56. goffarti a Sturhan 1966 (Transferred to Bitylenchus)
57. goldeni Rashid & Singh 1982 (syn. of T. elegans
58. gossypii Nasira & Maqbool 1996
59. graciliformis Siddiqi & Siddiqui 1983
60. haki Fotedar & Mahajan 1971
61. handooi Khan, A 2004
62. hexincisus Jairajpuri and Baqri 1968 (Transferred to Scutylenchus)
63. hordei Khan 1972
64. huesingi Paetzold 1958
65. iarius Saha, Gaur & Lal 1998
66. ibericus Mahajan & Nombela 1986 (Transferred to Quinisulcius)
67. impar Ray & Das 1983
68. indicus (Siddiqi) Fortuner & Luc,1987 1960 (Transferred to Quinisulcius)
69. iphilus Minagawa 1995 (Transferred to Bitylenchus)
70. irregularis Wu 1969
71. ismaili Azmi & Ahmad 1991
72. kamlae Shaw & Khan 1996
73. kangwoensis Geraert, Choi & Choi 1990
74. karnalensis Saha, Singh, Lal & Kaushal 2002
75. kashmirensis Mahajan 1974
76. kegasawai Minagawa 1995
77. kegenicus Litvinova 1946
78. kidwaii Rashid & Heyns 1990 (Transferred to Bitylenchus)
79. labiatus (Jairajpuri, 1984) Siddiqi 1986

50
 Check-list of globally known species of Tylenchorhynchus

[Link] Species Author Year Remarks

80. lamilliferus (de Man, 1880) 1936 (Transferred to
Neodolichorhynchus)
81. latus a Allen 1955
82. leucaenus Azmi 1991
83. leviterminalis Siddiqi, Mukherji & Dasgupta 1982
84. madrasensis Gupta & Uma 1981
85. malinus Lin 1992
86. mangiferae Laqman & Khan 1986
87. manubriatus Litvinova 1946
88. marudharensis Lal, Mathur & Rajan 1989 (Transferred to Telotylenchus
89. mashhoodi Siddiqi & Basir 1959
90. maximus Allen 1955 (Transferred to Sauertylenchus)
91. mexicanus Knobloch & Laughlin 1973
92. microcephalus Siddiqi & Patel 1990
93. microconus Siddiqi, Mukherjee & Dasgupta 1982
94. minutus Karpetjan 1979
95. musae Kumar 1981
96. namibiensis Rashid & Heyns 1990 (Transferred to Telotylenchus)
97. natalensis Kleynhans 1984 (Transferred to Bitylenchus)
98. neoclavicaudatus Mathur, Sanwal & Lal 1979
99. nilgiriensis Seshadri, Muthukrishnan & Shanmugam 1967 (syn. Quinisulcius capitatus)
100. nordiensis Khan & Nanjappa 1974
101. novenas Nobbs 1989 (Transferred to
Neodolichorhynchus)
102. nudus a Allen 1955
103. obscurisulcatus Andrassy 1959
104. oleraceae Gupta & Uma 1981
105. oryzae Kaul & Waliullah 1995
106. pachys Thorne & Malek, 1968
107. paracanalis Khan 1991
108. paranudus Phukan & Sanwal 1982 (syn. of T. leviterminalis)
109. paratriversus Brzeski 1991
110. parvus Allen 1955 (Transferred to Bitylenchus)
111. paulettae Bloemers & Wanless, 1998
112. penniseti Gupta & Uma 1980 (syn. of T. elegans)
113. persicus Sultan, Singh & Sakhuja 1991
114. phallocercus Chang 1991
115. phaseoli Sethi & Swarup 1968 (Transferred to
Neodolichorhynchus)
116. projectus Khan 1990
117. pruni Gupta & Uma 1981
118. punensis Khan & Darekar 1979 (syn. of T. elegans)

51
Sudershan Ganguly et al.

[Link] Species Author Year Remarks

119. punici Gupta and Uma 1980 (syn. Quinisulcius)
120. qasimi Ramzan, Handoo & Fayaz 2008
121. quaidi Golden, Maqbool & Handoo 1987 (Transferred to Bitylenchus)
122. queirozi Monteiro & Lordello 1976 (Transferred to Bitylenchus)
123. robustus Thorne & Malek 1968
124. rosei Zarina & Maqbool 1991 (syn. of T. varicaudatus)
125. sabourensis Shaw & Khan 1997
126. sacchari Sivakumar & Muthukrishnan 1983 (syn. of T. elegans)
127. sanwali Kumar 1982
128. sculptus Seinhorst 1963
129. shimizui Talavera, Watanabe & Mizukubo 2001
130. shivanandi Shaw & Khan 1992
131. siccus Nobbs 1990
132. silvaticus Ferris 1963
133. solani Gupta & Uma 1982 (Transferred to Neodolichorhynchus
134. spinaceai Singh 1976
135. striatus Allen 1955
136. sudanensis (Decker, Yasin & El-Amin, 1975) Castillo, 1989
Siddiqi & Gomez-Barcina
137. swarupi Singh & Khera 1981 (Transferred to Bitylenchus)
138. tarjani Andrassy 1969
139. tenuicaudatus Wouts & Sher 1981
140. teeni Hashim 1981 (Transferred to Bitylenchus)
141. teres (Khan & Darekar, 1979) Siddiqi 1986
142. thermophilus Golden, Baldwin & Mundo-Ocampo 1994
143. tobari Sauer & Annells 1981 (Transferred to Bitylenchus)
144. triglyphus Seinhorst 1963
145. trilineatus Timm 1963
146. tritici Golden, Maqbool & Handoo 1987
147. tuberosus Zarina & Maqbool 1994
148. usmanensis Khurma & Mahajan 1987 (Transferred to Bitylenchus)
149. varainnus Mavlyanov 1978
150. varicaudatus Singh 1971
151. velatus Sauer & Annells 1981 (Transferred to Sauertylenchus)
152. ventrosignatus Tobar Jime´ 1969 (Transferred to Bitylenchus)
153. vishwanathensis Pathak & Siddiqi 1997
154. vulgaris Upadhyay, Swarup & Sethi 1972 (Transferred to Bitylenchus)
155. wilskii Kornobis 1980 (syn. of sauertylenchus maximus)
156. yugaensis Geraert, Choi & Choi 1990
157. zambiensis Venditti & Noel 1995 (Transferred to Bitylenchus)
158. zeae Sethi & Swarup 1968
(a = Reported from India).

52
 Table 2. Check list of Tylenchorhynchus species described from India, along with their type host, locality, Reference and Accession Number in National
Nematode Collection of India (NNCI).

[Link]. Species Author year Type host Type Locality Reference Type accession Remarks
no. in NNCI

1 alami Shaw & Khan 1996 Musa sp. Ranchi, Bihar Journal of Research - -
Birsa Agricultural
University 8 : 1-8
2 allii Khurma & 1987 Allium cepa Jalandhar , Panjab Indian Journal of - (syn. of Bitylenchus
Mahajan Nematology 17: 202-207 quaidi)
3 amgi Kumar 1981 Coffea arabica Chickmaglur Journal of Coffee - -
Research 11: 88-99
4 badliensis Saha & Khan 1982 Lycopersicon Rohtak Indian Journal of 1192-1202 -
esculentum Nematology 11: 205-211
5 berberidis Sethi and Swarup 1968 Berberis North -Western Nematologica 5: 73-77 27-30 (syn. with Nagelus
aristata India hexagramus)
6 bambusi Singh, Lal, 2010 Bamboo Assam Indian Journal of 2291-2298 -
Rathour & Nematology.40 : 216-220
Ganguly

53
7 bohrrensis Gupta & Uma 1980 Sugarcane Bohrr [Link] .Parasitology, - -
40: 423-427
8 brassicae Siddiqi 1961 Cauliflower Aligarh Zeitschrift Parasitenkunde, - -
and cabbage 21: 46-64
9 cacti Chawala, 1968 Cactus spp. - Labdev Journal of Science - (Transferred to
Bhamburkar, and Technology.10: 205-210 Quinisulcius)
Khan & Prasad
10 chonai Sethi & Swarup 1968 Bougainvillea Western India Nematologica 14:77-88 31-34 -
spp.
11 cicerus Kakar, Khan & 1995 Pigeon pea Assam Annals of Plant Protection - -
Siddiqi Science. 3:149-154
12 dactylurus Das 1960 Capsicum Hyderabad Zeitschrift fur parasitenkunde - -
annum 19: 553-605
13 digitatus Das 1960 Lactuca sativa Hyderabad Zeitschrift fur Parasitenkunde - -
19: 553-605
14 coffeae Siddiqi & Basir 1959 Coffee Plants Mysore Proceedings of 46th Indian - -
Science Congress P.15
Check-list of globally known species of Tylenchorhynchus
 [Link]. Species Author year Type host Type Locality Reference Type accession Remarks
no. in NNCI

15 crotoni Pathak & Siddiqi 1997 Croton sp. Udaipur Indian journal Nematology - -
27: 99-103
16 cuticaudatus Ray & Das 1983 Potato Cuttack Indian Journal of Nematology - (Transferred to
13: 16-25 Bitylenchus)
Sudershan Ganguly et al.

17 cynodoni Kumar 1981 Coffee soils South India Journal of Coffee Research - -
Cynodon dactylon 11: 88-99
18 delhiensis Chawla, 1968 Anana Karol Bagh Labdev Journal of Science - -
Bhamburkar, squamosa and Technology 68: 86-100
Khan & Prasad
19 elegans Siddiqi 1961 Grass Aligarh Z. Parasitenkunde 21: 46-64 - -
20 divittatus Siddiqi 1961 - Aligarh Z. Parasitenkunde 21: 46-64 - -
21 goldeni Rashid & Singh 1982 sugarcane Lucknow Indian Journal of 1572-74 (syn. T. elegans)
Nematology 12: 193-195
22 haki Fotedar & 1971 Brassica Srinagar Kashmir Science: 120 - -
Mahajan oleracea

54
23 hexincisus Jairajpuri & 1968 Citrus plants Gujrata Nematologica 14: 217-222 - (Transferred to
Baqri Scutylenchus)
24 indicus (Siddiqi) 1960 Grass Aligarh Nematologica 5: 73-77 - (Transferred to
Fortuner & Quinisulcius)
Luc,1987
25 iarius Saha,Gaur & Lal 1998 Rice IARI Farm Annals of Plant protection 1833-1844 -
Science 6: 63-65
26 impar Ray & Das 1983 Broad Bean OUAT Orchard, Indian Journal of Nematology - -
Orissa 13: 16-25
27 ismaili Ajmi & Ahmad 1989 Albizta Jhansi Indian Journal of Nematology - -
19(2): 279-282
28 kamlae Shaw & Khan 1996 Kenddiopspy- Jamalpur ,Bihar Journal of [Link]. - -
rosexsculpta Agriculture. University 8: 1-8
29 karnalensis Saha, Singh, 2002 Rice Karnal Annanls of Plant Protection 2124-2129 -
Lal & Kaushal Science,10(2): 355-359
30 Kashmirensis Mahajan 1974 Brassica Srinagar Proceedings of Helminth- 834-835 -
oleracea ological Society of Washington
41: 13-16
 [Link]. Species Author year Type host Type Locality Reference Type accession Remarks
no. in NNCI

31 leucaenus Azmi 1991 Leucaena Jhansi Current Nematology.2: 81-82 - -

leucocephala
32 madrasensis Gupta & Uma 1981 Nanja Herb Plant Madras Helminthologica 18: 53-59 - -
33 mangiferae Luqman & Khan 1985 Mangifera Meerut Indian Journal of 1588-1589 -
indica Nematology 15: 202-208
35 marudharensis Lal, Mathur 1989 Datepalm Bikaner ,Rajasthan Indian Journal of - (Transferred to
& Rajan Phoenix dactylifera Nematology 9: 51-54 Telotylenchus)
34 mashhoodi Siddiqi & Basir 1959 Sugarcane Coimbatore Proc.46th Indian Science - -
Congress. P.15
36 musae Kumar 1981 Musa spp Chickmaglur Journal of Coffee - -
Research 11: 88-89
37 nilgiriensis Seshadri, 1967 Brassica Madras Current Science 36: 551-553 - (syn. Quinisulcius
Mutukrishnan oleracea capitatus)
& Shanmugam
38 neoclavi Mathur, Sanwal 1979 Imported USA Indian Journal of 1291-1292 -

55
caudatus & Lal Potato tubers Nematology 8: 148-150
39 nordiensis Khan & 1974 Citrus Panipat Indian Journal of - -
Nanjappa nomen Nematology 2: 216
novum for T.
areolatus
Khan &
Nanjappa, 1971
40 oleraceae Gupta & Uma 1981 Brassica Panipat Revista Ibero - -
oleracea Parasitologia 42:289-292
41 oryzae Kaul & 1981 Prunus persica Srinagar Annals of Plant Protection - -
Walliullah Science 3: 155-157
42 pruni Gupta & Uma 1995 Prunus persica Srinagar Helminthologica 18: 53-59 - -
43 punici (Gupta & 1987 Punica Srinagar Indian Academy of Science (syn. Quinisulcius)
Uma, 1980) granatum Animal. Science 89: 415-418
Fortuner & Luc
44 paranudus Phukan & Sanwal 1982 Areca catechu Karim ganj assam Indian Journal of 1371-1372 (syn. of T.
Nematology 12: 383-385 leviterminalis)
45 penniseti Gupta & Uma 1980 Pennisetum Naurangabad , Indian Journal of - (syn. T. elegans)
Check-list of globally known species of Tylenchorhynchus

typhoides Haryana Parasitology 4: 157-159

 [Link]. Species Author year Type host Type Locality Reference Type accession Remarks
no. in NNCI

46 persicus Sultan,Singh 1989 Prunus persica Ludhiana, Panjab Indian Journal of Nematology - -
& Sakhuja 19: 215-222
47 phaseoli Sethi & Swarup 1968 Phaseolus Panjab Nematologica 14:77-78 35 (Transferred to
aconotifolius Neodoli-
chorhynchus)
Sudershan Ganguly et al.

48 punensis Khan & Darekar 1978 Solanum Nasik, Maharashtra Indian Journal of 942-946 (syn. of T. elegans)
melongena Nematology 8: 43-48
49 sacchari Sivakumar & 1983 Sugarcane Quddalore, Indian Journal of 1443 (syn. of T. elegans)
Muthukrishnan Tamilnadu Nematology12: 393-395
50 sanwali Kumar 1982 Brassica Lucknow Kanpur University Research - -
oleracea 1: 185-192
51 savourensis Shaw & Khan 1997 Blackgram Bihar - - -
52 shivanandi Shaw & Khan 1992 Sugarcane Nagaland . India Bulletin of Entomology 33: 713 - -
53 solani Gupta & Uma 1982 - - Indian Journal of - (Transferred to
Parasitology 5: 37-38 Neodoli-

56
chorhynchus)
54 Spinaceai Singh 1974 Spinacea Lucknow Indian Journal of Zootomy - -
oleracea 15: 187-192
55 Swarupi Singh & Khera 1981 Cauliflower Hoogly, West Bulletin of Zoological - (Transferred to
Bengal Survey of India 1: 25-28 Bitylenchus)
56 usmanensis Khurma & 1987 Cucunis melo Panjab Indian Journal of - (Transferred to
Mahajan Nematology 17: 202-207 Bitylenchus)
57 variacaudatus Singh 1971 Fern Hyderabad Journal of Helminthology - -
45: 353-369
58 vishwanathensisPathak & Siddiqi 1996 Chandni Udaipur Indian Journal 0f - -
(Jasmium Nematology 26(2): 274-277
grandiflorum
59 vulgaris Upadhyay, 1972 Zea mays IARI,FARM Indian Journal of 622-639 (Transferred to
Swarup & Sethi Nematology 2: 129-138 Bitylenchus)
60 zeae Sethi & Swarup 1968 Zea mays Sangrur, Panjab Nematologica 14: 77-88 36-37 -
 Table 3. Diagnostic characters of nominal species of Tylenchorhynchus Cobb,1913 described from India.

Species L a b c c‘ V% Lip Lip Frame- Stylet Lateral Tail Tail Tail Spicule Guberna-
(mm) ratio region annules work (µm) incisureannules shape terminus (µm) culum
Sclerotization (µm)

aerolatus 0.55-0.68 27-34.4 5-6.3 14-17 2.4-2.7 52.7-56.6 CNT 4 LSC 11-13 4 14-19 BLROU SMO 20 10
alami 0.61-0.70 26.3-33.3 4.75-5.64 12.5-16 3-4 54.5-57.1 CNT 3 WSC 19-21 4 25 CON SMO 17-18 9-10
allii* 0.54-0.68 27.1-33.9 5.3-5.8 11-16 2.7-3.6 51.9-55 OFF 6-7 MSC 15-17 4 34-47 SCYL SMO 23-24.6 10-12.3
amgi 0.67-0.75 33-38 4.3-4.7 16-17 3.7 56-59 CNT 0 LSC 19-20 4 16-20 SCYL SMO - -
badliensis 0.62-0.71 28-35 5.0-5.1 15-19 3-4 53-61 OFF 3 LSC 17-19 4 17-22 CON SMO 20-22 10-11
berberidis* .72-.95 26-31 4.7-6.0 13-16 2.0-3.2 53-57 CNT 2-3 LSC 28-36 5-6 40 Bluntly ROU SMO 23-27 12-14
bambusi 0.66-0.77 27.2-38.0 4.8-5.7 13.3-18.7 2.7-4.5 43.7-56.2 CNT 2-3 LSC 17-19 4 17-25 CLA SMO 21-24 8-13
bohrrensis 0.61-0.75 24-30 4.7-6.0 13-18 2-2.3 53-57 CNT 2-3 LSC 15-17 4 17-21 SCYL SMO 23-27 12-14
brassicae 0.58-0.72 26-35 5.0-6.0 14-27 1.9-3.0 52-58 OFF 4 LSC 16-17 4 18-33 CON SMO 18-21 9-11
cacti* 0.6-0.7 25-33 4-6 14-19 - 54-59 OFF 6 - 15-19 5 23 CON SMO - -
chonai 0.56-0.66 28-32 4.3-5.8 15-17 2.5 57-62 CNT 3 - 25-28 3 14 CON SMO 21 14

57
cicerus 0.5-0.6 28.7-34.0 5.1-5.9 12.5-15 - 53-58 SEM OFF 3 MSC 13-16 3 - - - 20-23 9-11
coffeae 0.57-0.63 15-17 3-3.3 - - 2 - 17-20 4 19 SCYL SMO - -
crotoni 0.53-0.58 26.5-31.9 4.7-5.7 14.318.1 - 55.6-57 OFF 4-5 MSC 13-18 4 Fine CON ANN - -
cuticaudatus*0.50-0.62 31-37 4.2-4.8 12.3-15 2.3-2.4 53-60 OFF 5-6 LSC 14-16 4 33-45 CYL SMO 23-26 9-14
cynodoni 0.63-0.68 31-34 4.7-5.5 13-15 4.8 52-61 CNT 0-1 LSC 13-15 4 20-26 CLA SMO 28 11
dactylurus 0.64-0.71 30.8-32.5 4.6-5.0 14.8-16 - 52-54 CNT 4 LSC 20-21 4 25 BLP-ROU SMO 22 13
delhiensis 0.6-0.7 24-33 5-6 13-18 4 47-58 ROU 2 INC 14-16 4 29 SCYL SMO - -
digitatus 0.67-0.72 24-27 4.7-5.2 14.0-16.5 - 52-73.7 CNT INC 20 4 - CYL SMO 23 13
divittatus 0.55-0.77 32-38 5.4-6.4 16-19 2.8 53-55 OFF 5 INC 16-17 3 21 SCLA SMO 17 8
elegans 0.56-0.70 25-29 4.4-5.2 14.5-16 3.0 54-55 CNT 3-4 LSC 15-17 4 20-26 SCYL SMO 22-26 10-13
goldeni* 0.57-0.82 21-40 4.5-5.6 13-15 - 55-59 CNT 2-3 - 16-19 4 19-36 SCYL SMO 22-26 10-13
haki 0.55-0.63 26-33 5.0-5.9 20-27 2.5-4.0 56-59 CNT 3 LSC 16-18 4 14-20 CON SMO 18-22 10-15
hexincisus* 0.85-1.08 32-39 6.0-8.2 18-23 - 52-55 OFF 6 LSC 17-19 6 - SCYL ANN 28-31 8-9
indicus* 0.5-0.67 24-33 4.3-5.7 11.7-16.2 - 53-57 OFF 7 - 14-16.5 4 - - - 22-24 11-12.5
Check-list of globally known species of Tylenchorhynchus
 Species L a b c c‘ V% Lip Lip Frame- Stylet Lateral Tail Tail Tail Spicule Guberna-
(mm) ratio region annules work (µm) incisureannules shape terminus (µm) culum
Sclerotization (µm)

impar 0.57-0.68 24-29 4.5-5.3 16-20 1.9-2.2 55-60 OFF 4 LSC 22-24 3 11-14 CYL SMO 23-26 13-15
Ismaili 0.46-0.66 27-33 3.8-5.5 18-26 2.5-2.6 54-61 OFF 4-5 LSC 17-20 4 14-17 SCYL ANN - -
kamlae 0.55-0.63 27.4-32.1 4.4-5.7 11.7-16.2 3-4.7 53-59 OFF 4 MSC 19-21 4 21 SCYL SMO 22 11.7
Sudershan Ganguly et al.

karnalensis 0.59-0.74 24-29 4.5-5.5 12.5-15.5 2.7 – 3.2 50-55 CNT 3 LSC 16-17 4 22-28 CYL SMO - -
kashmirensis 0.60-0.74 24-34 4.0-5.5 24-37 - 62-64 OFF 3 MSC 17-21 4 13-17 CON ANN 19-21 -
leucaenus 0.55-0.56 36-37 4.6-4.7 16-22 - 55-56 OFF 4-5 LSC 17-18 4 18-21 SCYL ANN 13.5 5
madrasensis 0.53-0.66 23-25 4.4-4.7 15 1.9-2.2 56-59 CNT 1-2 LSC 19-22 3 10-13 SHM SMO 22-24 12
mangiferae 0.54-0.67 32-39 5.0-5.7 14-16 - 51-56 OFF 5-6 - 12-13 4 35-38 SCLA SMO 22 7-9
mashhoodi 0.61-0.76 26-30 4.9-5.5 16-19 2.5-4.0 55-59 - 3-4 LSC 17-18 4 14-16 CYL SMO 22-24 12-13
marudharensis*0.64-0.81 36-42 6.0-6.6 14-18.5 3.0-3.7 52-55 OFF 7-8 LSC 14.5-16.0 4 35-42 CON SMO 23.5-25 11-13
musae 0.58-0.65 28.1-32.8 4.9-6.0 14.8-16.4 - 54.9-56.9- OFF 1-2 LSC 18-19 4 16-20 SCYL SMO 24-26 10-12
nilgiriensis* - - - - - - - - - - - - - - - -

58
neoclavic- 0.59-0.72 23-31 4.0-5.8 12-15 27-36 53-56 CNT 2-3 LSC 20-23 4 32-50 CLA SMO 23-25 11-15
audatus
nordiensis 0.55-0.68 27.4-34.4 5.0-6.3 14-17 - 52.7-56.6 CNT 4 LSC 11-13 4 14-19 ROU SMO 20 10
oleraceae 0.48-0.68 - - 12-16 2.2-3.2 - OFF 4-5 - 12-15 4 38 CON SMO - -
oryzae 0.43-0.61 24-29 3.6-5.1 12.8-18.7 - 53-62 OFF LSC 20-23 4 >20 SCYL SMO 20-24 11-12.5
pruni 0.58-068 25-30 5.0-5.9 16-22 1.7-2.6 55-62 OFF 4-5 LSC 14-17 3 17 CON SMO 24 9
punici* 0.65-0.76 - - 15-18 2.1-3.2 - OFF 6-7 - 16-17 5 38-42 CON SMO - -
paranudus 0.59-0.78 25-30 4.8-5.8 12-16 2.5-3.7 51-56 OFF O - 18-21 4 14-25 CLA SMO 23-27 12-14
penneseti* 0.53-0.67 - - - 2.0-3.2 - CNT 3 - 15-17 4 13-17 CYL SMO - -
persicus 0.71-0.73 35-41 4.9-5.8 13-17 - 50-55 OFF - MSC 14-16.5 - - CON BLUNT 22-24 12-14
phaseoli* 0.61-0.77 28-35 4.0-5.4 17-20 54-58 OFF 6 LSC 19-26 4 22 CON SMO 23.5 11.5
punensis* 0.60-0.75 31-41 4.6-5.5 12-15 3.7-4.8 53-59 CONROU 2-3 WSC 16-18 4 24-27 CYL SMO 20-22 9-10
sacchari* - - - - - - - - - - - - - - - -
sanwali 0.45-0.71 21.5-23.7 7.5-10.3 16.3-18.7 2 - OFF 2 - 19.5 4 30-31 CYL ANN - -
savourensis 0.49-0.70 30-36 4.8-5.5 13-17 3-3.5 53-58 SEMOFF 5 LSC - 3 20 - SMO 21-22 8-9
 Species L a b c c‘ V% Lip Lip Frame- Stylet Lateral Tail Tail Tail Spicule Guberna-
(mm) ratio region annules work (µm) incisureannules shape terminus (µm) culum
Sclerotization (µm)

shivanandi 0.48-0.64 26-35 4.5-5.7 12-16 3-4 52-59 CNT 4 MSC 16-19 4 21 SCYL SMO - -
solani* 0.60-0.70- - - - - - - 5-6 - 16-17 4 34 CON SMO - -
spinaceae 0.60-0.81 - - - - - - 4 - 14-18 4 12-18 CYL SMO - -
swarupi* 0.42-0.54 - - 14-15 2.9 - OFF 5-6 - 13-15 4 - CON - - -
usmanensis 0.55-0.65 25.7-30.2 5.0-5.6 12.2-14 2.3-3.0 52.7-56 OFF 5-6 LSC 14-16 4 34-40 CON SMO 16-24 11-11.5

59
varicaudatus 0.50-0.56 28-33 4.3-5.2 16-19 2.3-2.8 57-59 CNT 2 LSC 17-18 4 14-15 CON SMO - -
vishwanathensis0.53-0.67 25-26.6 4.8-5.7 9.5-28.8 - 55.4-57.8 OFF - MSC 15-20 4 FINE CON ANN - -
vulgaris* 0.56-0.67 25-30 4-6 14-20 - 52-57 OFF 6-7 MSC 14-16 4 35-42 CON SMO 22-25 13-16
zeae 0.53-0.64 26-34 4.8-6.0 14-20 2.6 57-61 CNT 4 LSC 17-20 4 16 CON SMO 20 9

Shape of lip region: CNT= Continuous; CON = Conoid; OFF = Offset; SEM-OFF = Semi-offset; ROU = Round.
Frame-work Sclerotization: LSC= Lightly Sclerotized; MSC = Moderately Sclerotized; WSC = Weakly Sclerotized; INC = Inconspicuous.
Shape of tail: BLR = Bluntly- rounded; CLA = Clavate; CON = Conoid; CYL = Cylindrical; SCLA = Sub-clavate; SCYL = Sub-cylindrical; ROU = Rounded; BLP = Bluntly- pointed;
SHM = Semi-hemispherical.
Shape of tail terminus: SMO= Smooth; ANN = Annulated.
* Transferred/ Synonymized with other genera/ species.
Check-list of globally known species of Tylenchorhynchus
Sudershan Ganguly et al.

ACKNOWLEDGEMENT Agriculture Nematode Collection, Beltsville, Maryland.

Journal of Nematology 30: 108-158.
Authors are thankful to the Head, Division of
Hooper, D.J. (1978). The Tylenchorhynchidae. The identification
Nematology and Director, IARI, New Delhi for providing
of stunt nematodes (Tylenchorhynchinae, Merlininae and
facilities. Thanks are also due to Mr. Siddappa Annigeri, Trophurinae), especially those in western Europe. Pp. 1–
Ph.D. student for his help in procuring the literature. 21 in spiral and stunt nematodes. Manual prepared for
workshop sponsored by the Nematology A Check.
REFERENCES
Jairajpuri, M.S. (1982). Some studies on Tylenchorhynchinae:
Allen, M.W. (1955). A review of the nematode genus
The subgenus Bitylenchus Filipjev, 1934, with description
Tylenchorhynchus. University of California
of Tylenchorhynchus (Bitylenchus) depressus n. sp. and
Publications in Zoology 61: 129-166.
a key to species of Bitylenchus. Mededelingen van de
Anderson, R.V. & Potter, J.W. (1991). Stunt nematodes: Fakulteit Landbouwwetenschappen, Rijksuniversiteit
Tylenchorhynchus, Merlinius, and related genera. Pp. Gent 47: 765-770.
529–586 in W. R. Nickle, ed. Manual of Agricultural
Mahajan, R. (1988). A conspectus of the genus
Nematology. New York: Marcel Dekker.
Tylenchorhynchus Cobb, 1913 (Nematoda:
Brzeski, M.W. & Dolinski, C.M. (1998). Compendium of the Tylenchorhynchinae). Indian Journal of Nematology
genus Tylenchorhynchus Cobb, 1913 sensu lato 18: 199-206.
(Nematoda: Belonolaimidae). Russian Journal of
Minagawa, N. (1995). Bitylenchus iphilus sp. n. and
Nematology 6: 189-199.
Tylenchorhynchus kegasawai sp. n. (Nematoda:
Cobb, N.A. (1913). New nematode genera found inhabiting Tylenchida) from Japan. Afro-Asian Journal of
fresh-water and non-brackish soils. Journal of the Nematology 5: 151-160.
Washington Academy of Sciences 3: 432-445.
Powers, T.O. (1983). Systematic analysis of Merlininae Siddiqi,
Esser, R.P. (1991). A computer-ready checklist of the genera 1971 (Nematoda: Dolichodoridae). Ph.D. thesis.
and species of phytoparasitic nematodes, including a University of California, Riverside.
list of mnemonically coded subject categories. Bulletin
Powers, T.O., Baldwin, J.G. & Bell, A.H. (1983). Taxonomic
13. Florida Department of Agriculture and Consumer
limits of the genus Nagelus (Thorne and Malek, 1968)
Services, Division of Plant Industry, Contribution No.
Siddiqi, 1979 with a description of Nagelus borealis n.
387, Gainesville.
sp. from Alaska. Journal of Nematology 15: 582-593.
Filipjev, I.N. (1934). Harmful and useful nematodes in rural
Siddiqi, M.R. (1986). Tylenchida parasites of plants and insects.
economy. Moscow, Leningrad. (In Russian.)
Commonwealth Agricultural Bureaux, Slough, London,
Fortuner, R. & Luc, M. (1987). A reappraisal of Tylenchina United Kingdom. Pp. 645.
(Nemata). 6. The family Belonolaimidae Whitehead,
Siddiqi, M.R. (2000). Tylenchida: parasites of plants and
1960. Revue de Nematologie 10: 183-202.
insects. Commonwealth Agricultural Bureaux, Slough,
Golden, A.M., Maqbool, M.A. & Handoo, Z.A. (1987). London, United Kingdom. Pp. 833
Description of two new species of Tylenchorhynchus
Sitaramaiah, K. (1984). Plant parasitic nematodes of India.
Cobb, 1913 (Nematoda: Tylenchida), with details of
Today & Tomorrow’s Printers & Publishers, Karol
morphology and variation of T. claytoni. Journal of
Bagh, New [Link]. 292.
Nematology 19: 58-68.
Swarup, G., Dasgupta, D.R. & Koshy, P.K. (1989). Plant diseases.
Gomez-Barcina, A., Siddiqi, M.R. & Castillo, P. (1992). The genus
Anmol Publications, Darya Ganj, New Delhi. Pp. 395
Bitylenchus Filipjev, 1934 (Nematode: Tylenchida) with
descriptions of two new species from Spain. Journal of the Tarjan, A.C. (1964). A compendium of the genus
Helminthological Society of Washington 59: 96-110. Tylenchorhynchus (Tylenchidae: Nematoda).
Proceedings of the Helminthological Society of
Handoo, Z.A. (2000). A Key and Diagnostic Compendium to
Washington 31: 270-280.
the Species of the Genus Tylenchorhynchus Cobb, 1913
(Nematoda: Belonolaimidae). Journal of Nematology Tarjan, A.C. (1973). A synopsis of the genera and species in
32: 20-34. the Tylenchorhynchinae (Tylenchoidea, Nematoda).
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Washington 40: 123-144.
specimens on deposit in the United States Department of

60
 Efficacy of carbofuran against root-knot nematode on tomato
Vol. 43, No. 1, pp. 61-64 Indian Journal of Nematology June, 2013

Role of Organic Carbon on the Efficacy of Carbofuran Against Root-Knot

Nematode on Tomato
H.K. SHARMA, PANKAJ AND JAGAN LAL

Division of Nematology, Indian Agricultural Research Institute, New Delhi-110 012, India
*Corresponding author, E-mail:

Received on 02-04-2013 and Accepted on 21-04-2013

ABSTRACT: Investigation was carried out to study the efficacy of carbofuran (1 and 2 kg a.i/ha) with the varying levels of FYM 5,10
and 20%w/w in tomato infected with root-knot nematode under green house conditions. Results on number of root-knot galls
indicated that carbofuran efficacy increased with the levels of FYM. Plant growth such as shoot wt. and shoot length improved in
treatment while root wt. was decreased amongst treatments.

Key words: Root-knot nematode, tomato, FYM, carbofuran

Nematicides form an important component of significance of organic matter coupled with carbofuran
nematode management, applied to soil against soil born in the management of important nematode pest like root-
nematodes parasitic to crops. These chemicals come in knot nematode infecting tomato has been investigated to
contact with soil particles as well as with organic matter study the role of organic matter for the better efficacy of
present in soil. Thus, affecting the overall activity of carbofuran.
biological entities in soil. The quality of organic matter
also has implication on chemical activity. Together, soil MATERIALS AND METHODS
particle do affect the chemical dynamics of the compound.
The organic matter and chemical interaction phenomenon An experiment was conducted to study the role of
involved adsorption and desorption of chemical compound farm yard manure (FYM) on the efficacy of carbofuran
which determine the persistence and decomposition of against root-knot nematode, Meloidogyne incognita
molecule. This determine availability of chemical in soil infecting tomato in 15 cm earthen pots in completely
water interphase. It is well known that there is a direct randomize design with three replication under green
correlation of amount of organic matter and persistence house conditions. Healthy seedlings of tomato cv. Pusa
of chemical compound and also amount of organic Ruby were raised in sterilized soil and sand mixture (1:1)
matter has inverse relation with uptake of chemical by in earthen pan. Sterilized sandy loam soil was filled in
plant. Thus, quantity of FYM plays a major role in the each pot and thirty days old seedlings were planted in
pesticide efficacy. Beside, it may also affect the pesticide pots. FYM was applied @ 0, 5, 10 and 20% w/w per pot
by having good growth of soil microflora that also has at the time of filling the pots. Freshly hatched juveniles
bearing on pesticide. The systemic nematicides are (J2s) were collected from infected tomato roots,
generally adsorbed on FYM which can affect the maintained in the Division of Nematology, IARI. These
availability of pesticide in a soil environment (Bansal, J2s were inoculated @ 2 J2/g soil after 7 days of
2009, Bansal, 2010). The organic matter and pesticide transplant. Subsequently, carbofuran 3G @ 1 and 2 kg
interaction is well reviewed by Gabriela et al., 2007) a.i./ha was applied per pot 24h after nematode inoculation.
Management of root knot nematode quite often is achieved Inoculated control was kept as check. Observations on
with the chemical nematicides like carbofuran. The plant growth parameters (such as root and shoot weight

61
H.K. Sharma et al.

and shoot length), nematode multiplication (number of than higher level of carbofuran. In general, there was
galls and egg mass) and soil population were observed significant reduction in number of galls by different
after 60 days of treatment. Soil population was assessed treatments in comparison of control. The number of gall
by washing 200 cc soil collected from the rhizosphere of reduced by carbofuran 1 kg a.i/ha at FYM 20% do not
the treated and control plants. The soil was processed as vary significantly with carbofuran 2kg a.i./ha at any level
per the procedure by Cobb (1918). Further, reproduction of FYM.
factor was calculated by using formula Pf/Pi (Pf= final
nematode population, Pi= Initial nematode population). Similar observations were recorded on number of
egg mass and soil population of nematode in various
RESULTS AND DISCUSSION treatments. Reproduction factor was lower in combined
use of carbofuran and FYM than the carbofuran alone
Number of galls caused by root-knot nematode in application.
tomato under different treatments indicated that efficacy
of carbofuran was altered by varying dose of FYM With regards to plant growth of tomato i.e shoot wt.
(Table 1). Root-knot galls progressively decreased with enhanced with both the concentrations of carbofuran
an increase of FYM (5-20%) with carbofuran 1 kg a.i/ with the rise in FYM levels. The increase was observed
ha, as the nematode galls decreased from 30 to 82% over to be about 72% at low levels of carbofuran over control.
control. However, in comparison to carbofuran alone the Combined application of FYM and carbofuran improved
reduction ranged between 5 to 74% clearly suggests, the the shoot wt compared to control and when carbofuran
role of FYM in influencing the efficacy of chemical. At was applied alone. Similar observation was made at
higher carbofuran concentration, reduction in nematode higher level of carbofuran. The shoot weight improved
galls was 79% which further increased to 84% at highest by the application of carbofuran alone compared to
level of FYM over control. Similarly, in contrast to control. Shoot length also showed similar pattern as that
carbofuran 2 kg a.i./ha the reduction in nematode galls of shoot wt. Shoot length was not significantly affected
between ranged of 15-22%. Therefore, significant effect in carbofuran 1 kg a.i/ha applied in 10 and 20% FYM or
of FYM on the carbofuran efficacy against nematode carbofuran 2 k.g a.i/ha in FYM 5%. Shoot length show
observed at lower concentration of carbofuran rather
Table 2. Effect of carbofuran in presence of varying dose of
Table 1. Effect of carbofuran in presence of varying dose of FYM against root knot nematode infected tomato
FYM on root knot nematode multiplication on tomato. plant growth.

Treatments Number Number Soil Treatments Shoot Root Shoot

of of egg population/ wt wt length
galls mass 200cc soil (g) (g) (cm)

Carbofuran 1kg 220 40 400 Carbofuran 1kg 15.2 5.1 23.3

Carbofuran 1kg a.i/ha+ FYM 5% 200 30 250 Carbofuran 1kg a.i/ha+ FYM 5% 11.9 5.0 22.3
Carbofuran 1kg a.i/ha+FYM 10% 110 20 100 Carbofuran 1kg a.i/ha + FYM 10% 18.9 4.5 30.8
Carbofuran 1kg a.i/ha+FYM 20% 57 10 100 Carbofuran 1kg a.i/ha FYM 20% 19.2 3.6 30.5
Carbofuran 2kg 77 12 300 Carbofuran 2kg 15.8 3.9 30.5
Carbofuran 2kg a.i/ha+ FYM 5% 50 15 100 Carbofuran 2kg a.i/ha+ FYM 5% 21.8 3.5 27.0
Carbofuran 2kg a.i/ha+FYM 10% 65 8 150 Carbofuran 2kg a.i/ha + FYM 10% 26.0 4.4 39.5
Carbofuran kg a.i/ha+ FYM 20% 52 7 100 Carbofuran kg a.i/ha+ FYM 20% 25.4 4.1 32.0
Control 320 60 500 Control 11.4 5.7 18.8
CD (P=0.05) 11.3 CD (P=0.05) 2.6 0.6 3.3

62
 Efficacy of carbofuran against root-knot nematode on tomato

carbamates was more in unamended soil compared to

manure amended soil and decreased with the increase in
amount of organic manure (Bansal, 2010). Adsorption of
carbamate pesticides by clays have shown that they are
adsorbed by co-ordination and/or protonation at the
carbonyl oxygen by exchangeable cations of clays (Bansal
1983, 2009; Li et al., 2003). Shamalie et al. (2011)
reported no effect of organic matter on the efficacy of
carbofuran either on plant growth or nematode
development, may be because doses of carbofuran and
organic matter were too low. The high amount of residue
of carbofuran linked to high amount of organic matter
(Khuntong et al., 2010) and at high pH degradation is
faster. This has implication on nematode control. In the
[Link] 1kg, [Link] 1kg a.i/ha+ FYM 5%,
[Link] 1kg a.i/ha+FYM 10%, [Link] 1kg a.i/ present study carbofuran 2 kg a.i/ha do not have effect
ha+FYM 20%, [Link] 2kg, [Link] 2kg a.i/ha+ varied with organic matter probably that this concentration
FYM 5%, [Link] 2kg a.i/ha+FYM 10%, [Link] kg
was too high for organic matter (5-20%) to hold this and
a.i/ha+ FYM 20%, [Link]
thus more carbofuran might be available in soil water
Fig. 1. Reproduction factor of root knot nematode as phase that is why it is more effective against nematode.
influenced by different treatment on tomato
The study find that carbofuran efficacy increased with
FYM dose point out that carbofuran was adsorbed more
significant improvement with an increase in FYM levels and released for longer period by FYM and hence its
at both the concentrations of carbofuran. Maximum effect as nematocide was more pronounced. Carbamate
plant growth was recorded in carbofuran 2 kg a.i/ha and pesticides has been reported to be adsorbed by organic
FYM (10-20 %) treatment. Improvement was more than material amended soil may be retained for a longer time
71%. Shoot length increased with the increase in FYM in the soil and control the pesticidal activity effectively
levels. (Bansal, 2010). This property also has significance on
the efficacy of pesticides and that suggest that it is better
In contrast, root weight of tomato decreased with the
to apply pesticide as per the FYM levels in soil. It could
increase in FYM doses with carbofuran 1kg a.i/ha while
be inferred that FYM does play a role in the effectiveness
at higher dose (2 kg a.i/ha) of carbofuran there had been
of carbofuran against root-knot nematode and
little improvement even though it was lower than control.
accordingly reflected in plant growth as well. Beside,
Root weight was not affected significantly in the
shoot weight and length improved in treatments over
treatments however in comparison to control it was
control but root weight was lower in treatments in
affected.
comparison to control.
The variation in root-knot galls in tomato plant in
REFERENCES
response to systemic nematicide has to do with adsorption
and desorption of chemical on organic matter which Bansal, O.P. (1983). Adsorption of oxamyl and dimecron in
actually suggest whether the chemical is available in soil montmorillonite suspensions and desorption of three
water phase to be effective against pest. (Durovic et al., carbamate pesticides. Soil Science Society of America
2009;Osborn et al., 2009; Villavarde et al., 2008). The Journal 47: 877-883.
adsorption increased with the increase in organic manure.
The adsorption capacity was significantly positively Bansal, O.P. (2009). Adsorption and desorption of three
carbamate pesticides by montmorillonite and humic
correlated with soil organic carbon and CEC and
acid-clay complexes. Journal of Indian Society of Soil
negatively correlated with soil pH. Desorption of systemic Science 57: 287- 290.

63
H.K. Sharma et al.

Bansal, O.P. (2010). The Effects of composts on adsorption- Khuntong, S., Sanya, S., Pakkong, P. & Cheema, S. (2010).
desorption of three carbamate pesticides in different Adsorption kinetics of carbonate pesticide in rice fed
soils of Aligarh district. Applied Sciences Environment soil. Environment Asia 3: 20-28.
Management 14: 155-158.
Li, H., Sheng, G., Teppan, B.J., Johnston, C.T. & Boyd, S.A.
Durovic, R., Gajic-umiljendic, J. & Dordevic, T. (2009). Effects (2003). Sorption and desorption of pesticides by clay
of organic matter and clay content in soil on pesticide minerals and humic acid- clay complexes. Soil Science
adsorption processes. Pesticides Phytomedicine 24: Society America Journal 67: 122-131.
51-57.
Osborn, R.K., Edwards, S.G., Wilcox, A. & Haydock, P.P.J.
Fernandes, M.C., Cox, L., Hermosin, M.C. & Corenjo, J. (2009). Potential enhancement of degradation of the
(2003). Adsorption-desorption of metalaxyl as affective nematicides aldicarb, oxamyl and fosthiazate in U.K.
dissipation and leaching in soils: role of mineral and agricultural soils through repeated applications. Pest
organic components. Pest Management Science 59: Management Science 66: 253-261.
554-552.
Shanalie, B.V.T., Fonseka, R.M. & Rajapaksha (2011). Effect
Gabriela, B., Graciela, P. & Nelson, D. (2007). Influence of of Trichoderma viride and carbofuran (curator) on
organic amendment on the biodegradation and movement management of root-knot nematode and growth
of pesticides. Environmental Science and Technology parameters of gotukola (Centella asiatica L.). Tropical
37: 233-271. Agricultural Research 23: 61-69.

64
 Evaluation of Paecilomyces lilacinus in flue cured virginia (FCV) tobacco nursery
Vol. 43, No. 1, pp. 65-69 Indian Journal of Nematology June, 2013

Evaluation of Paecilomyces lilacinus for the Management of Root-Knot

Nematode, Meloidogyne incognita in Flue Cured Virginia (FCV)
Tobacco Nursery
S. RAMAKRISHNAN* AND C. PANDURANGA RAO

Central Tobacco Research Institute Research Station, Hunsur-571105, Karnataka, India

*Corresponding author and E-mail: ramkictri@[Link]

Received on 10-04-2013 and Accepted on 15-05-2013

ABSTRACT: Flue Cured Virginia (FCV) tobacco is a major commercial crop grown in light soil regions of Karnataka. Root-knot
nematodes, Meloidogyne incognita is a major threat to the successful production causing heavy yield and quality loss. Nematicide
like carbofuran is being used against this nematode, but with limited efficacy. Extensive use of pesticides of chemical origin
especially in higher doses for nematode control has to be avoided due to higher costs and associated hazards to the environment.
Replicated trials were conducted with varied dosage levels of Paecilomyces lilacinus against the nematode in FCV tobacco nursery
raised in root-knot nematode sick plots. Results revealed that at the time of final pulling, P. lilacinus @ 30g/m2 recorded significantly
increased total healthy transplants count of 857.5/m2 and was on par with P. lilacinus @ 25g/m2 (845.1/m1. Similarly at 60 DAS,
applications of P. lilacinus @ 30g/m2 and @ 25g/m2 were on par with each other in recording the significantly reduced RKI of 1.81
and 1.88 respectively. Both the treatments were also on par with combined application P. lilacinus @ 30g/m2 + Carbofuran @ 10g/
m2, which recorded the significantly reduced RKI of 1.81 compared to untreated check (3.86).

Key words: Paecilomyces lilacinus, carbofuran, Meloidogyne incognita, FCV tobacco nursery.

Flue-Cured Virginia (FCV) tobacco is an important caused by Fusarium oxysporum f. sp. nicotianae
commercial crop grown in Karnataka light soils (KLS) as contributing to significantly reduced yields. Both fumigant
rainfed crop has lot of export potentiality. KLS tobacco and non-fumigant nematicides such as dazomet and
is preferred internationally due to its ideal chemistry with carbofuran have been successfully used against this
below detectable levels of TSNA (Tobacco specific nematode in nursery (Ramakrishnan et al., 1998). But
nitrosamines) compounds and pesticides residues. Among excessive use of synthetic pesticides is not cost effective
the various pest and diseases associated with the crop, and moreover many effective nematicides were
plant parasitic nematodes and particularly the root-knot withdrawn from the market due to their ill-effects and
nematode, Meloidogyne incognita pose serious threat hazards they pose to environment. FCV tobacco. grown
for the production and productivity causing significant in Karnataka being an exportable commodity, presence
reduction in terms of both yield and quality of the tobacco of chemical residues is a great concern and hence use of
crop in nursery and main field to the tune of 59.4% and such nematicides is highly discouraged. Bio-management
52.9% respectively (Hussaini, 1983; Ramakrishnan et of root-knot nematode with antagonistic organisms is an
al., 2001). Root-knot nematode infested seedlings, when alternative, cost effective and eco-friendly approach.
transplanted in main field, exhibit stunted growth and Biological control of plant parasitic nematodes is important
may even collapse tesulting in gaps. Losses caused by in view of long-term advantage of management.
this nematode are very high, especially when they interact Paecilomyces lilacinus is the potential egg parasitic
with other disease causing pathogens. Ramakrishnan et fungus, which is highly safe and has been used successfully
al. (2008) had reported that root-knot nematode, M. to manage root-knot nematodes in various crop plants
incognita predispose FCV tobacco crop to wilt disease (Ekanayake & Jayasundara, 1994; Jonathan et al., 1995;

65
S. Ramakrishnan and C. Panduranga Rao

Ramakrishnan & Nagesh, 2011). The fungus affects the dimension of 100 cm2, from which mean was calculated.
egg masses, engulfs and penetrates the eggs and Twenty five seedlings were examined for root-knot
proliferates within them by consuming the egg contents infection per replication in each treatment and graded
(Zaki & Bhatti, 1990). Hence, an attempt was made to under 0-5 scale, from which mean RKI was calculated.
evaluate commercial formulation of Paecilomyces In addition, number of galls/g. root, egg mass/g. root and
lilacinus (2 x 106 spores/g) in varied dosage levels and eggs/egg mass were also recorded. Soil samples were
also in combination with carbofuran under replicated drawn before the application of treatments (initial) and at
trials for the management of root-knot nematode, M. the end of the experiment (final) for the estimation of
incognita in FCV tobacco nursery. infective nematode population in the soil. Data gathered
were statistically analysed using standard procedure.
MATERIALS AND METHODS
RESULTS AND DISCUSSION
The replicated nursery trials for two seasons were
conducted during 2011 & 2012 in the root-knot nematode All the treatments evaluated under nursery conditions
infested nursery site of CTRI Research Stattion, Hunsur. were found to be significantly superior over check in both
Raised nursery beds each of 1.2 m X 1.2m size were reducing the root-knot disease incidence and in
prepared to fine tilt and the mean initial population of subsequently improving the seedling growth and root
infective juveniles of M. incognita was 171 J2/100g soil. knot free seedlings count. Data from Table 1 revealed
The commercial formulation of the egg parasitic fungi, that, there was no adverse effect of the product, Jai V Jai
Jai V Jai -P. lilacinus formulated as wettable powder -P. lilacinus on FCV tobacco seed germination. At 45
with a spore load 2 x 106 spores/g formulations was DAS, P. lilacinus @30g/m2. significantly differed from
incorporated into the raised nursery beds as per treatment the other dosage levels and increased the mean healthy
schedule. The treatments incorporated includes, Jai V transplantable seedlings count to 316.5/m2 compared to
Jai -P. lilacinus at varied dosage levels (ie) @ 5, 10, 15, 216.0/m2 in untreated check. Whereas, at 60 DAS, P.
20, 25 & 30g/m2, Neem cake @ 400g / m2 + Soil lilacinus @ 30g/m2 recorded maximum number of mean
Solarization, carbofuran @ 10 g/m2, Jai V Jai -P. lilacinus healthy transplantable seedlings count (320.3/m 2)
@ 30g / m2 + carbofuran @10g / m2 and with one compared to check (237.1/m2). But, at the time of final
untreated check. The treatments were replicated thrice pulling, P. lilacinus @ 30g/ m2 recorded significantly
in Randomised Block Design. For the treatment Neem increased mean total healthy transplants count of 857.5/
cake @ 400g / m2 + Soil Solarization, where soil m2, which is 53.7 per cent increase over check. It was
solarisation was involved, after the incorporation of also on par with P. lilacinus @ 25g/m2, which recorded
neem cake, prepared nursery beds were irrigated and mean total healthy transplantable seedlings count of
covered with clear low density polyethylene (LDPE) 845.1/m2, which is 51.5 per cent increase over untreated
film of 25 μm thickness when the moisture levels in the check (557.8/m2). Similar yield increase in okra plants
beds were around field capacity. Edges of the sheets grown in root-knot sick soil due to soil application of P.
were sealed with mud and left undisturbed for six weeks lilacinus was reported by Dhawan et al. (2004). Similar
period. Then the sheets were removed and after two to to the present results, Nagesh et al. (2001) had obtained
three days of waiting period, FCV tobacco seed of significantly increased yield of chrysanthemum flowers
variety “Kanchan”, susceptible to root-knot nematode to the tune of 23 -28% by use of talc-formulation of P.
was sown at the rate 0.3g/ m2 in all the nursery beds. All lilacinus @ 4 -6 kg/ac against root-knot nematodes. In
the other agronomic practices were followed as the present investigations, carbofuran @ 10g/m2, the
recommended. Observations such as Germination count recommended nematicide increased the total healthy
(at 10 DAS), Count of healthy transplants at 60 DAS and transplantable seedlings count to the tune of 22.8 per
at final pulling, root knot index (RKI) at 60 DAS and at cent only compared to check. Whereas its combined
final pulling were recorded. Germination count was application with P. lilacinus @ 30g/m2 recorded the
taken at 15 DAS at random in ten squares, each with maximum total healthy transplants count to the tune of

66
 Evaluation of Paecilomyces lilacinus in flue cured virginia (FCV) tobacco nursery

Table 1. Effect of Paecilomyces lilacinus on root-knot free and healthy transplants counts in FCV tobacco nursery.

Treatment details Germ. Healthy % inc. Healthy % inc. Total % inc.

count transplants over transplants over healthy over
count check count check transplants check
(45 DAS) (60 DAS) count

Paecilomyces lilacinus (2 x 106 21.1 235.6 9.0 292.8 23.5 658.0 18.0
cfu/g) @5g/m2
P. lilacinus (2 x 106 cfu/g) @10g/m2 21.8 240.0 11.1 281.5 18.7 655.5 17.5
P. lilacinus (2 x 106 cfu/g) @15g/m2 21.6 235.1 8.8 294.0 23.9 682.3 22.3
P. lilacinus (2 x 106 cfu/g) @20g/m2 20.3 273.6 26.6 313.1 32.1 759.1 36.1
6 2
P. lilacinus (2 x 10 cfu/g) @25g/m 21.2 307.5 42.3 336.1 41.7 845.1 51.5
P. lilacinus (2 x 106 cfu/g) @30g/m2 21.1 316.5 46.5 320.3 35.1 857.5 53.7
Neem cake @400g/m2 21.7 278.0 28.7 303.0 27.7 676.0 21.1
+ Soil Solarization
Carbofuran @10g/m2 21.2 279.0 29.1 295.5 24.6 685.0 22.8
P. lilacinus (2 x 106 cfu/g) @30g/m2 20.5 313.1 44.9 336.8 42.0 860.8 54.3
+ Carbofuran @ 10g/m2
Control 20.3 216.0 - 237.1 - 557.8 -
[Link] 0.78 1.84 3.92 3.16
CD (P=0.05) NS 5.10 10.86 8.77

54.3 per cent and it was on par with both the best dosage DAS, application P. lilacinus @ 30g/m2 in FCV tobacco
levels (25 &30 g/m2) of P. lilacinus. nursery beds significantly reduced root-knot index (RKI)
to 1.30 on 0-5 Scale, which is 50.1% decrease over
At the time of final pulling, data from table 2 revealed untreated check (2.61) and was found on par with P.
that application of P. lilacinus @ 30 g/m2 and 25 g/m2 lilacinus @ 25g/m2 (1.31). Similarly at 60 DAS,
were on par with each other in significantly reducing the applications of P. lilacinus @ 30g/m2 and @ 25g/m2
number of egg mass/g. root to the tune of 31.0 and 26.5 were on par with each other in recording the significantly
per cent respectively and eggs/egg mass to the tune of reduced RKI of 1.81 and 1.88 respectively. Decrease
27.0 and 24.6 per cent respectively and final soil over untreated check was 53.1 and 51.3 per cent
population/100 g soil to the tune of 52.5 and 49.6 per cent respectively. Both the best treatments were also on par
respectively compared to un treated check. In general, with combined application P. lilacinus @ 30g/m2 +
the application of P. lilacinus at varied dosage levels Carbofuran @ 10g/m2, which recorded the significantly
from 5 to 30g/m2 in FCV tobacco nursery against root- reduced RKI of 1.81 compared to untreated check
knot nematodes caused 16.5 to 31.0 per cent reduction (3.86). Similarly Reddy & Khan (1988) reported 76%
in number of egg mass/g root, 13.7 to 27.0 per cent reduction in population of reniform nematode,
reduction in number of root-knot nematode eggs/egg Rotylenchulus reniformis with combined application of
mass and 30.2 to 52.5 per cent reduction in final soil carbofuran at 2 kg a.i/ha and P. lilacinus @ 2g on
nematode population as compared to un treated check. tomato. Saikia & Das (2001) got highest reduction in
Similar decrease in root-knot nematode soil population galls (60%) with combined application of P. lilacinus @
due to application of P. lilacinus in banana was also 2g/kg soil and carbofuran @ 1kg. This clearly exhibits
reported earlier by Jonathan & Rajendran (2000). At 45 the compatibility of P. lilacinus with the chemical

67
 Table 2. Effect of Paecilomyces lilacinus on root knot nematode multiplication in FCV tobacco nursery .

Treatment details RKI at % RKI at %Dec. [Link] % Dec. [Link] % Dec. Soil % Dec.
45 Dec. 60 over Egg over Eggs/egg over population over
DAS over DAS check masses check mass check (/100 g. check
S. Ramakrishnan and C. Panduranga Rao

check /g. root Soil)

Paecilomyces lilacinus (2 x 106 cfu/g) @5g/m2 2.30 11.8 3.03 14.5 16.3 18.5 250.0 14.8 99.5 31.8
P. lilacinus (2 x 106 cfu/g) @10g/m2 2.38 8.8 3.13 18.9 16.7 16.5 253.0 13.7 101.8 30.2
6 2
P. lilacinus (2 x 10 cfu/g) @15g/m 2.33 10.7 2.95 23.5 16.0 20.0 245.5 16.3 101.0 30.8
P. lilacinus (2 x 106 cfu/g) @20g/m2 2.30 11.8 2.60 32.6 15.4 23.0 242.5 17.4 87.3 40.2

68
P. lilacinus (2 x 106 cfu/g) @25g/m2 1.31 49.8 1.88 51.3 14.7 26.5 235.0 19.9 73.5 49.6
6 2
P. lilacinus (2 x 10 cfu/g) @30g/m 1.30 50.1 1.81 53.1 13.8 31.0 214.1 27.0 69.3 52.5
Neem cake @400g/m2 + Soil Solarization 2.26 13.4 2.45 36.5 16.5 17.5 242.5 17.3 93.8 35.7
2
Carbofuran @10g/m 2.00 23.4 2.25 41.7 15.7 21.5 242.0 17.5 82.6 43.4
6 2
P. lilacinus (2 x 10 cfu/g) @30g/m + 1.27 51.3 1.81 53.1 13.5 32.5 214.5 26.9 69.0 52.7
Carbofuran @ 10g/m2
Control 2.61 - 3.86 - 20.0 - 293.5 - 146.0 -
[Link] 0.06 0.03 0.54 1.62 1.80
CD (P=0.05) 0.16 0.08 1.49 4.49 4.98
 Evaluation of Paecilomyces lilacinus in flue cured virginia (FCV) tobacco nursery

nematicide, carbofuran. Moreover, both the best incognita in chrysanthemum using formulations of
treatments in the present study were also significantly Paecilomyces lilacinus in combination with neem cake.
In: Proc. 2nd National symposium in Integrated Pest
superior to sole application of carbofuran @ 10g/m2 with
Management in Horticultural Crops- New Molecules,
RKI of 2.25 in reducing the root-knot disease incidence Biopesticide & Environment” IPM in Horticultural
in FCV tobacco nursery. The experimental results clearly Crops: Emerging Trends in the New millennium (Eds.
indicate that P. lilacinus @ 30g/m2 was on par with P. A. Verghese and P .Parvatha Reddy), AAPMHE, UHR,
lilacinus @ 25g/m2 in reducing the root knot nematodes Bangalore. pp. 149-150.
incidence in FCV tobacco nursery and in subsequently
increasing the total root-knot free and healthy seedlings Ramakrishnan, S., Hussaini, S.S., Viswanath, S.M. & Shenoi,
M.M. (1998). Effect of Basamid G for the control ofroot-
count. Hence, it is concluded that, application of P.
knot nematodes in FCV tobacco nursery. In: Proc. Nat.
lilacinus in talc-formulation with spore load of (2 x 106 Symp. Rational approaches in nematode management
cfu/g) @ 25g/m2 (ie) 3 kg/unit nursery (120m2) is an ideal for sustainable agriculture. (Eds. Dhawan, S. C and
dosage for the effective management of root-knot Kaushal, K. K). GAU, Anand, pp. 121-124.
nematodes in FCV tobacco nursery.
Ramakrishnan, S., Sreenivas, S.S. & Viswanath, S.M. (2001).
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Ramakrishnan, S., Shenoi, M.M. & Sreenivas, S.S. (2008).
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Ramakrishnan, S. & Nagesh, M. (2011). Evaluation of
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reniformis infecting tomato, compared with carbofuran.
Jonathan, E.I., Padmanabhan, D. & Ayyamperumal, A. (1995).
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Management of root knot nematode, Meloidogyne

69
V. Bamel and R.K. Sonkar
Vol. 43, No. 1, pp. 70-73 Indian Journal of Nematology June, 2013

Effect of Phorate 10G and Neem Cake on Tylenchulus semipenetrans in a

Declining Nagpur Mandarin Orchard in Central India
V. BAMEL* AND R.K. SONKAR

National Research Centre for Citrus, Amravati Road, Nagpur – 440 010, India
Corresponding author, E-mail: vbamel@[Link]

Received on 15-04-2013 and Accepted on 17-05-2013

ABSTRACT: A field experiment was conducted during the year 2008-10 to assess the effect of phorate 10G @ 1.0, 1.5 and 2.0 kg a.i.
per ha and neem cake @ 10, 15, 20 kg/ tree on population behavior of Tylenchulus semipenetrans in a declining Nagpur mandarin orchard
in central India. Nematode population levels were assessed before treatment and after 4, 8 and 12 months following treatment.
Application of phorate 10G and neem cake beneath the tree canopy resulted in significantly higher % reduction (44.2% in soil and
48.4% in roots in case of phorate 10G @ 2.0 kg / ha and 39.2% in soil and 32.8% in roots in case of neem cake @ 15 kg/ tree) in nematode
population compared to untreated control plants (22.6% in soil and 20.59% in roots) twelve months after treatment. The results
indicated that in general the percent reduction over initial population was significantly more in trees treated with phorate 10G
in comparison to trees treated with neem cake. Percentage reductions in population of females per gram fresh root weight were
approximately double in plants receiving phorate 10G compared to untreated control plants. Plants receiving phorate 10G supported
minimum nematode reproduction (Rf = 0.55) compared to all treatments and control plants. Maximum increase in canopy volume
(9.54%) was observed in the treatments where phorate 10G was applied @ 2 kg per ha.

Key Words: Tylenchulus semipenetrans, Nagpur mandarin.

In India, main fruits of Citrus group include lime, available nematicide, phorate 10 G and neem cake were
lemon, mosambi and orange (mandarin). Nagpur mandarin tested for controlling the population of citrus nematode in
(Citrus reticulata Blanco) is a major fruit crop grown in an established mandarin orchard heavily infested with
central India. In India, Citrus is cultivated over an area this nematode.
of 846 thousand hectares with a total production of 7464
thousand MT and productivity of 8.8 MT/ ha during the MATERIALS AND METHODS
year 2011 (Anon, 2011). The citrus nematode,
Tylenchulus semipenetrans, has been a problem in both Investigations were carried out on a sixteen-years-
established and replanted orchards and its infection leads old Nagpur mandarin orchard (block No. 30) with a total
to slow decline of citrus (Du Charme, 1969). Rough area of about 2304 m2 situated at National Research
lemon (Citrus jambhiri), the most commonly used Centre for Citrus, Nagpur with Citrus jambhiri (rough
rootstock for Nagpur mandarin is highly susceptible to lemon) rootstock. The orchard was on drip irrigation
citrus nematode. Survey of various citrus growing areas system. Trees were growing at a spacing of 6x6 m.
of Vidarbha showed that Citrus nematode (T.
semipenetrans) is a major nematode problem of Nagpur Soil samples were collected individually from all 64
mandarin orchards which require immediate attention plants of the block. Each composite sample consisted of
for its management (Bamel 2006 and 2009). Several four sub-samples from four sides beneath the canopy of
management options have been tried in India (Ahmad, each tree at a distance of 1.5 m from tree trunk and up
1985; Alam et al. 1977, Mani et al. 1986). Commonly to 40 cm depth in the fibrous root zone with the help of

*Present Address: Division of Nematology, Indian Agricultural Research Institute, New Delhi – 110 012

70
 Effect of phorate 10G and neem cake on Tylenchulus semipenetrans

soil auger. Each composite sample was mixed well and Nematode population levels in the soil as well as roots
a 100 cc working sample was drawn and washed as per were assessed after 4, 8 and 12 months following
Cobb’s sieving and decanting technique and further treatment. The treatments and observations were
processed for 48 h on a modified Baermann funnel. The repeated after one year.
J2s of T. semipenetrans were collected after 48 h and
counted under stereoscopic binocular microscope. Fibrous The canopy volume was calculated as per the
roots collected from each composite sample were washed formula π/ 6 x HD2, where H is the height of the plant
and processed to recover females of T. semipenetrans. above tree trunk and D is the average diameter of the
One gram roots from each composite sample were canopy in East-West and North-South direction
weighed and stained in boiling cotton blue lectophenol for
30 seconds and then placed in plain lectophenol for 48 h. RESULTS AND DISCUSSION
Females were counted under stereoscopic binocular
microscope. Reproduction factor was calculated by The results presented in Table 1 clearly indicated
dividing the nematode final population in soil and roots by that application of phorate 10G and neem cake beneath
the initial nematode population in soil and roots. the tree canopy resulted in significant reduction in
nematode population compared to untreated control
Trees were treated during the month of December plants. The results indicated that in general the percent
when nematode population is much higher (Singh, 1997) reduction over initial population was significantly more in
with phorate 10G @ 1.0, 1.5 and 2.0 kg a.i. per ha and trees treated with phorate 10G in comparison to trees
neem cake @ 10, 15, 20 kg/ tree beneath the canopy treated with neem cake. Four months following treatment,
about 1 m away from tree trunk and in a band of about maximum reduction in soil (73.2%) and root (74.1%)
1 m wide in the feeder root zone and incorporated population of the nematode were recorded in plants
mechanically after applying light irrigation. All sides of receiving phorate 10G @ 2.0 kg a.i. per ha. With an
tree were treated, and untreated trees served as controls. increase in dose of phorate 10G, there was a
corresponding higher decrease in soil as well as root
All the treatments were replicated 8 times and population. In case of neem cake, the % reduction in
arranged in a randomized complete block design. population was found to be significantly less compared to

Table 1: Effect of phorate 10G and neem cake on canopy volume and population of Tylenchulus semipenetrans in soil and roots
at 4, 8 and 12 months after treatment in Nagpur mandarin

Treatment *% reduction in nematode population after

4 months 8 months 12 months % Increase in
Soil Root Soil Root Soil Root canopy volume
(/100cc) (/g) (/100cc) (/g) (/100cc) (/g) (m3) after two
years

Phorate 10 G @1 kg a.i./ ha 62.4 55.4 67.3 61.4 37.4 38.5 4.93

Phorate 10 G @1.5 kg a.i./ ha 67.4 67.7 70.0 66.5 39.5 41.5 6.74
Phorate 10 G @2.0 kg a.i./ ha 73.2 74.1 73.6 71.1 44.2 48.4 9.54
Neem cake @ 10 kg/ tree 53.7 39.5 39.8 33.6 31.9 26.0 4.87
Neem cake @ 15 kg/ tree 54.3 42.7 44.4 36.4 35.5 31.5 4.06
Neem cake @ 20 kg/ tree 62..8 47.5 51.9 41.0 39.2 32.8 6.87
Control 40.2 28.2 34.4 22.8 22.6 20.59 2.96

(*Pooled data of 2008-09 and 2009-10)

71
V. Bamel and R.K. Sonkar

trees receiving Phorate 10G after twelve months of a corresponding decrease in nematode reproduction
application (39.2% in soil and 32.8% in roots in case of factor. Maximum value of reproduction factor was
neem cake @ 15 kg/ tree and 44.2% in soil and 48.4% observed in control plants (Rf = 0.78). At the end of
in roots in case of phorate 10G @ 2.0 kg / ha). Reduction second year, maximum increase in canopy volume in the
in root population showed similar trend to soil population. treatments was observed where phorate 10G was applied
The reduction was also observed in soil (40.2%) and @ 2 kg per ha (9.54%) followed by neem cake @ 20 kg
roots (28.2%) after four months in control plants. This per tree (6.87%). During the course of experimentation,
may be probably due to natural decline of nematode the plants were not given the stress treatment for
population during summer (Singh, 1997). The estimation flowering. Fruit set due to natural flowering were recorded
of nematode population after 8 month of treatments and found to be non-significant.
showed that the nematode population decreased further.
However, the % reduction in population decreased with Singh, B. (2004) observed in a field trial for the
time. There was a decrease in soil and root population control of the citrus nematode, Tylenchulus
after one year of treatment in all the treatments and semipenetrans in a ten year old Nagpur mandarin
population were lower compared to initial population. orchard on rough lemon rootstock that the application of
Subsequent measurements demonstrated that application carbofuran 3G and phorate 10G, each at 1, 3 and 5 kg a.i/
of nematicide or neem cake reduced population levels to ha reduced the nematode (Tylenchulus semipenetrans)
approximately half the level of those in untreated control. populations in soil and on roots significantly within one
Phorate 10G or neem cake applied under canopy reduced month of nematicide application. The repeated application
adult female nematode population. Twelve months after of the nematicides after one year kept the namatode
treatment, percentage reduction in population of females populations significantly lower as compared to non-
per gram fresh root weight were approximately double in repeated and control treatments. Tiwari & Vadhera
plants receiving phorate 10G at all doses compared to (1999) found that application of phorate @ 4 kg a.i./ ha
control plants. Nematode reproduction factor at the end reduced 48% nematode population in roots and increased
of second year (Fig. 1) showed that plants receiving the plant yield by 49% compared to untreated plants. Mc
phorate 10G supported minimum nematode reproduction Clure & Scmitt (1996) also observed the effect of Rugby
(Rf = 0.55) compared to other treatments. With an 10G up to 12 months for suppression of the citrus
increase in dose of nematicide or neem cake, there was nematode in lemon trees. Philis (1993) observed the
effective control of nematodes and increase in grapefruit
yield by 74.2% and 53.2% with fenamiphos and aldicarb,
respectively. Philis (1997) achieved effective control of
nematode with application of Rugby (cadusafos) for
consecutive three years.

It may be concluded that application of phorate 10G @

2.0 kg/ ha may be used for reducing the population of citrus
nematode in citrus orchards, as the chemical reduced the
nematode population in soil (44.2%) as well as roots
(48.4%) and increased the canopy volume by 9.54%.

Rf: Reproduction factor; T1: Phorate 10 G @1 kg a.i./ ha; T2:

ACKNOWLEDGEMENTS
Phorate 10 G @1.5 kg a.i./ ha; T3: Phorate 10 G @2.0 kg a.i./ ha;
T4: Neem cake @ 10 kg/ tree; T5: Neem cake @ 15 kg/ tree; T6: We are grateful to Dr. Pankaj, Principal Scientist,
Neem cake @ 20 kg/ tree; T7: Control Division of Nematology, IARI, New Delhi for critical
Fig. 1: Effect of Phorate 10G and neek cake on comments on the manuscript and Director, National
reproduction factor of Tylenchulus semipenetrans Research Centre for Citrus, Nagpur, for providing all the
infecting Citrus reticulata (Blanco) facilities.

72
 Effect of phorate 10G and neem cake on Tylenchulus semipenetrans

REFERENCES McClure, W.A. & Schmitt, M.E. (1996). Control of citrus

nematode, Tylenchulus semipenetrans with cadusafos.
Anonymous (2001). Indian Horticulture Database published Journal of Nematology 28 (4), Supplement: 624-628.
by National Horticulture Board. pp 44-47.
Philis, I. (1993). Control of the citrus nematode, Tylenchulus
Ahmad, N.S. (1985). Studies on citrus nematode, Tylenchulus semipenetrans in established grapefruit orchard in
semipenetreans Cobb with reference to distribution, Cyprus. Nematology 21: 63-66.
biology and control. M. Sc. (Ag.) Thesis. A. P. Agriculture
University, Hyderabad, pp.84. Philis, I. (1997). Effect of citrus nematode control on the yield
and fruit quality grapefruit in Cyprus. Miscellaneous
Alam, M.M., Siddiqui, S.A. & Khan, A.M. (1977). Use of Reports-Agricultural Research Institute, Ministry of
organic amendments in nematode management. Indian Agriculture and Natural Resources, Nicosia. No. 66 p.
Journal of Nematology 7: 27-37. 6.

Bamel, V. (2006). Phytonematodes associated with Nagpur Singh, B. (1997). Population fluctuation of Tylenchulus
mandarin orchards in central India. Proceedings of semipenetrans Cobb in Vidarbha region of Maharashtra.
National Symposium on Citriculture: A Road Map held Pest Management in Horticultural Ecosystem 3: 53-56.
at ICAR Research Complex for NEH Region, Umiam
during 22nd to 24th February, 2006. Singh, B. (1999). Occurrence of distribution of Tylenchulus
semipenetrans Cobb in Nagpur mandarin orchards of
Bamel, V. (2009). Community analysis of plant parasitic Vidarbha, Maharashtra, Indian Journal of Nematology
nematodes associated with Nagpur mandarin orchards 29: 149-153.
in Wardha district (Maharashtra). In: Proceedings of
National Symposium on Citriculture: Emerging Trends Singh, B. (2004). Control of citrus nematode, Tylenchulus
(24-26July 2008), NRCC, Nagpur. semipenetrans in Nagpur mandarin orchard. Indian
Journal of Nematology 34: 70-74.
Du Charme, E.P. (1969). Nematode problems of citrus
(Peachey,J.E. ed). In: Nematodes of Tropical Crops, Tiwari, S.P. & Vadhera, I. (1999). Prevalence of citrus
Common Wealth Agriculture Bureau, pp.225-237. nematode, Tylenchulus semipenetrans in Madhya
Pradesh and its management in citrus orchard. Journal
Mani, A. Ahemed, B.N., Kameshwara Rao, P. & of Mycology and Plant Pathology 29: 11-15.
Dakshinamurti, V. (1986). Plant products toxiz to the
citrus nematode, Tylenchulus semipenetrans ,Cobb.
Int. Nematology Network Newsletter 3: 14-15.

73
C. Bhattacharya et al.
Vol. 43, No. 1, pp. 74-81 Indian Journal of Nematology June, 2013

Biodiversity of Plant Parasitic Nematodes in Tea Nurseries and

Plantations in Tripura
C. BHATTACHARYA*, M.K. DASGUPTA** AND B. MUKHERJEE*** (LATE)

* ‘Banalata’ , Ramnagar Road No -6, P .0. -Ramnagar, Agartala, Tripura (West) -799002, India
** Visva-Bharati, Palli-Siksha Bhavana (Institute of Agriculture), Department of Plant Protection, Sriniketan 731236 West Bengal, India
Present Address :’Oikos, Simantapalli, Santiniketan -731235, West Bengal, India
***Netaji Subhash Mahavidyalaya, Udaipur, Tripura, India
*Corresponding author, E-mail: bhattacharyachitra@[Link]

Received on 15-04-2013 and Accepted on 20-05-2013

ABSTRACT: One hundred and thirty sites at thirteen widely separated tea nurseries and plantations of different age groups were
sampled for qualitative and quantitative analysis of plant parasitic nematodes communities during March to October, 2001. A total
of 14 species of plant parasitic nematodes within eight genera, were isolated and identified from roots and rhizosphere soil of tea
plants. All species of plant parasitic nematodes associated with tea are first records of nematodes associated with tea from Tripura.
The most frequently encountered nematode species includes: Helicotylenchus spp., Meloidogyne spp., Pratylenchus brachyurus (Pb ),
Rotylenchulus reinformis (Rr), Hoplolaimus spp. and Hemicriconemoides mangiferae (Hm), which were detected in 85%, 70%, 68%, 55%,
53% and 41% of the sites. respectively. Nematode communities were analysed by mean abundance, prominence value, importance
value and diversity index based on nematode numbers and biomass. Analysis showed that Meloidogyne hapla ranked first in
importance value followed by M. incognita,H, dihystera R. reniformis and H. erythrinae. A linear increase in the index of diversity (H’)
of the nematode communities with increase in the age of plantation up to eight years was observed and with the further increase
in the age of plantation there is a gradual decline in the diversity index. This indicated that with an increase in the age of plantations
up to certain age parasitic species have become more abundant. This is a distinct sign of gradual establishment of these species.

Key words: Biodiversity index, Community analysis, Plant parasitic nematode, Tea, Tripura, India.

Tea (Camellia sinelisis L.) occupies a prominent against the all India average 1000 kg/ ha (Bhattacharjee
place as a cash crop as well as an export crop in India. & Singh, 1995)
In Tripura, there are 59 active tea plantations (Numbers
mentioned in parenthesis with each sub-division) : Agartala Nematode damage to tea plantations in India has
Sadar (25), Kailasahar (16), Dharmanagar (8), Kamalpur been well documented only from Darjeeling District,
(5), Khowai (2) Sabroom (2) and Belonia (1), subdivisions West Bengal (Mukherjee and Dasgupta, 1982) and
of the state (Anonymous, 2001). The agroclimatic catelogued 19 species of plant parasitic nematodes
conditions in Tripura are suitable for development of tea within 13 genera from the state. The serious pathogenic
plantations. The fertile soil, without major problems of nematodes of tea including Pratylenchus loosi, P.
toxicity or deficiency, high annual rain fall (2400mm) brachyurus, Meloidogyne brevicauda and
makes Tripura the fifth largest in average among the 14 Helicotylenchus erythrinae were detected from the
tea producing states after Assam, West Bengal, Tamil soil and root samples. They concluded that with an
Nadu and Kerala. This is cultivated in the high slopes, increase in altitude, diversities of nematode communities
hillocks and in plain lands with diversified agroclimatic declined, Association of P. loosi with devastation and
conditions covering a land area of 6124,22 ha. under the debilitations in Sri Lanka. Japan and Bangladesh have
cultivation. The average production of tea is 2749 kg/ ha been noted (Campos et al., 1990). Records of plant

74
 Plant parasitic nematodes in tea nurseries and plantations in Tripura

nematode infestation of tea plantation in Assam included hydrogen peroxide (Gowen & Edmunds, 1973) for
Meloidogyne incognita, M. hapla and P. brachyurus extraction of migratory endoparasites. Batches of five
(Phukan, 1998). Subsiquently M. brevicauda has been gms of tender roots were stained by acid fuchsin
recorded from matured tea plants in Coonor, Tamil Nadu lactophenol method and nematode population in roots
(Mehta & Somasekhar, 1998). Although a large number was estimated after extraction through maceration in a
of plant parasitic nematodes have been encountered blender. Plant parasitic nematodes were identified up to
from soil around the tea plants, the known and suspected genus and species level in permanent mounts, in
pathogens are Pratylenchus spp., Meloidogyne spp., dehydrated glycerol. The species of root-knot nematodes
Radopholus similis, Helicotylenchus spp., was identified on the basis of perinneal pattern, male and
Rotylenchulus reniformis, Hoplolaimus spp., juvenile characters (Eisenback, 1985).
Rotylenchus spp., Xiphinema spp. and
Hemicriconemoides spp. through out the world (Koshy, Population density (mean ± SE) with range and
2002). Since no information is available about the plant frequency of ocurrence of each nematode genus was
parasitic nematodes associated with tea in Tripura, an determined at each location and for total samples collected
intensive nematological survey for qualitative and from the area. Nematode cornrnunites were analysed by
quantitative analysis of pJant parasitic nematodes prominence value (Absolute density X V Absolute
associated with tea nurseries and plantations situated in frequency) and importance values (Relative density =
widely separated geographical locations in the state. Relative frequency + Relative biomass) for each nematode
species following the formula given by Norton (1978).
MATERIALS AND METHODS Biomass was determined by morphometric measurement
of 20 females collected from each location.
During March to October, 2001, an intensive survey
of plant parasitic nematodes asociated with tea plants Shannon -Weiner diversityindex, H’ =L Pilog Pi,
was conducted in nurseries at 3 locations (6 months to 2 where’s’ is the number of species at each location and
years old) and plantations at 10 locations (3 to 25 years ‘Pi’ is the relative abundance of the ‘i’th species, was
old) in the main tea producing areas in Tripura state. A used to determine the diversity of nematode community.
total of 130 composite soil and root samples were Diversity of nematode species was measured by Shannon-
collected. At each location, 10 sites were fixed oil a given Weiner information measure to base ‘e’.
sampling date. At each site, within a radius of 100 meters,
20 mature tea bushes were randomly selected and soil Physicochemical analysis of soil samples
and root samples were collected. Each sample consisted
of 20 sub-samples of rhizosphere soil and associated The representative soil samples collected from the
roots in a sample area of 30m2 of nursery and 1ha. of root zones of tea at different locations were labelled and
plantation up to 40 cm depth with the help of a GI pipe transported to the laboratory .These samples were air
(2.5 cm dia). The age of the nurseries and plantations dried in the lahoratory for 10 days, grinded and passed
were recorded as reported by the estate managers and through coarse sieves and then stored in polythene bags.
government officials at the tea estate concerned. Mechanical analysis of soil samples were carried out by
International pipette menthod (Piper, 1966) to differentiate
Nematodes were extracted from 250 cm3 composite the sand, silt and clay content (%). Determination of pH
soil sub-samples by modified Baermann funnel technique of solid samples were conducted by a glass electrode pH
(Whitehead & Beming, 1965). The root samples were meter (Blackman) as described by Jackson (1967).
washed in running tap water, cut into smaller pieces, Determination of organic carbon was done by Walkely
which were thoroughly mixed. Five gm of these composite and Black’s rapid titration method (Piper, 1966).
root masses were macerated in 100 ml tap water in a Estimation of available nitrogen by alkaline permanganate
blender for two min. The resultant root suspensions were method, available phosphorus byBray and Kurtz method
incubated for three days over double layered facial tissue and available potassium by Flame Photometric method
paper on coarse wiremesh placed in water using 1-3% (Jackson, 1967) were carried out.

75
C. Bhattacharya et al.

RESULTS plant parasitic nematodes associated with tea plants are

being reported for the first from Tripura.
Fourteen species of plant parasitic nematodes within
eight genera were isolated and identified from roots and The Physicochemical properties of tea soils showed
rhizosphere soils around tea plants. The species were that the soils were sandy clay type in all locations, acidic
Helicotylenchus dihystera, (Cobb, 1893) Sher, 1961, (pH 4.3-6.8) with high clay (34-65%), higher sand (24-
H. microcephalus Sher, 1966, H. erythrinae 48%) and low silt (9-17%) content. Organic carbon
(Zimmerman, 1904) Golden, 1965, Hemicriconemoides varied from 0.38 to 1.5% and a wide variation in NPK
mangiferae, Siddiqi, 1961, Hoplolaimus columbus, status (Table 1). Diversity of nematode community
Sher, 1963 H. indicus Sher, 1963, Meloidogyne hapla based on nematode numbers (H’n) was highest in the
(Chitwood 1949). M. incognita (Kofoid & White, 1919) plantation located at Mahespur (8 years old), than either
Chitwood, 1949. Pratylenchus brachyurus (Godfrey, the younger or older plantations (Fig. 1). According to
1929) Filip jev & Sch. 1941, Rotylenchulus reniformis Niblack and Bernard (1985), correlations were found
Linfored & Olivera, 1940 Tylenchorhynchus between sand, silt or clay content of soil and densities of
brevilineatus Williams, 1960, T. Mashhoodi Siddiqi & several nematode species but densities of these species
Basir, 1959 Xiphinema elongatum Sch. Stek & were not related to soil texture class.
Teunissen, 1983 and X, insigne Loos , 1949. Among
these, Helicotylenchus spp. (85%), Meloidogyne spp. Nematode abundance
(70%), Pratylenchus brachyurus (68%),
Rotylenchulus reniformis (55%), Hoplolaimus spp. The mean density and frequency of occurrence of
(53%) and Hemicriconemoides mangiferae (41%) were different nematode differed in nurseries and plantations.
most abundant and predominant nematodes effecting The spiral nematodes, Helicotylenchus spp. were
both nurseries and plantations (Table 1). All species of represented by three different species, viz. H. dihystera,

Table 1: Physicochemical properties of tea soils in Tripura1

Location Sand Silt Clay Organic Organic Nitrogen Phosphorus Potassium pH

(%) (%) (%) matter (%) carbon (%) kg/ha kg/ha kg/ha

Nursery
Durgabari 30 10 34 0.654 0.380 0.03 0.5 11 6.2
Harendranagar 25 9 47 0.909 0.529 0.05 0.5 12 6.4
Gokulnagar 27 9 51 0.923 0.537 0.05 0.6 11 6.2
Plantation
Mekhlipara 48 12 37 1.245 0.724 1.3 1.5 15 6.1
Durgabari 43 12 46 0.964 0.561 0.05 0.7 16 4.6
Maheshpur 34 14 53 1.252 0.728 0.04 0.6 17 6.7
Sabroom 24 11 65 1.985 1.154 0.03 0.5 12 6.5
Harendranagar 40 17 43 0.938 0.545 1.2 1.0 15 4.4
Mahabir 43 11 47 1.378 0.811 0.07 0.3 25 6.3
Golakpur 37 11 52 1.203 0.7 0.06 0.2 24.4 5.7
Lakhilunga 48 14 38 2.027 1.185 0.06 0.3 14 6.8
Gokulnagar 44 16 40 0.929 0.540 0.5 1.6 17.4 4.3
Rangung 40 11 49 2.56 1.5 0.8 0.5 22 6.3
1
Soil type (Sandy clay in all locations)

76
 Plant parasitic nematodes in tea nurseries and plantations in Tripura

lesions on the epidermal layer of the feeder roots indicated

the involvement of the pathogen in different plantations.
The mean abundance of P. brachyurus was highest (98
nematodes/g) on root at Harendranagar (12 years old)
and the population ranged from 53 to 118 nematodes per
g of root samples. The species has been recorded long
back from tea plants of the adjoining state of Assam
(Basu, 1968) and from Darjeeling, West Bengal
(Mukharjee & Dasgupta, 1982).

The reniform nematode, Rotylenchulus reniformis,

was present both from nurseries and plantations. The
mean density of R. reniformis was 88 nematodes
ranging from 28 to 320 nematodes per 250 cm3 soil in
nursery. The species was also uniformly present in
different plantations except Mekhlipara although the
density varied. The mean density of the species was 104
nematodes per 250cm3 soil in plantations, which was
Fig. 1. Community analysis showing importance values and much higher than that in nurseries (Table 1). The reniform
their components for 14 plant parasitic nematodes
nematode was much more prevalent in nursery plants,
associated with tea in Tripura state, India
causing 100% infection by the juveniles and young
females which were observed in soil samples too. The
H. erythrinae and H. microcephalus, among which H. same species has caused casualities in young tea fields
dihystera was the most prominent. The mean densities in Indonesia and widely prevalent in different tea estates
of the special nematodes, Helicotylenchus spp. were 93 of Sri Lanka in the elevation range 200 to 900m amsl
and 182 nematodes per 250 cm3 soil and the population (Campos et al., 1990).
ranged from 10 to 189 per 250 cm3 in nurseries and 12 to
516 in plantations respectively. Three nurseries (6 months to 2 years old) and ten
plantations (3-25 years old) surveyed varied greatly with
Root-knot nematodes, Meloidogyne spp. were respect to age and in nematode abundance. Population
uniformly present in all nurseries and plantations surveyed. densities of all identified species also varied widely
The frequency was 80% in nurseiies and 70% in among nursery sites and plantations (Table 1).The
plantations. Their population ranged from 6 to 272 (mean incidence of Hemicriconemoides mangiferae,
96) nematodes in nurseries and 10-376 (mean 124) Hoplolaimus indicus, H. columbus, Tylenchorhynchus
nematodes in plantations. Root-knot disease caused by brevilineatus, T. mashoodi, Xiphinema elongatum
M. incognita and M. hapla were most common in and X. insigne were observed in nurseries and plantations
nursery and young plants below 5 years of age. The sizes alike in Tripura. The population load of different nematode
of the root galls varied widely in different plantations. pathogens at higher age groups of tea plantations become
The larger sized root-galls (2-16 mm dia) were recorded less severe because with the increase in age of the
in younger plantations (3-5 years of age) at Mekhlipara plants, the tannin content of the roots increased. The
and Durgabari in West Tripura district. The sympatric concomitant occurrence of M. incognita, M. hapla,
prevalence of M. hapla and M. incognita in tea roots Pratylenchus brachyurus and R. reniformis suggested
were found. Lesion nematode, Pratylenchus brachyurus a multiple species infection in Tripura. It appears that the
was more abundant in plantations (53 nematodes) than in root-knot nematodes and lesion nematodes are a growing
nurseries (34 nematodes per 250 cm3 soil). The frequency threat and a potent major constiaint in tea nurseries of the
of occurrence was also higher (68%) in plantations than state, where sanitation and othe management practices
in nurseries (50%). The presence of dark red cortical are advisable.
77
C. Bhattacharya et al.

Nematode community structure

Nematode communities were analysed by absolute

density, relative density, absolute frequency, relative
frequency, prominence value, importance value and
diversity index. Analysis of the nematode communities
showed that M. hapla ranked first in importance value
(70.64) followed by M. incognita (60.85), H. dihystera
(33.27), R. reniformis (22.77) and H. erythrinae (19.28).
As the importance values of M. hapla and M. incognita
were more than two to three times those of other
prominent nematode species viz. H. dihystera R. Fig. 2. Nematode community diversity in relation to age of
reniformis and H. erythrinae. However. H. dihystera tea plantations in Tripura, India
occupied the first rank in relative density (18.50) and
relative frequency (14.56) followed by M. incognita the plantation located at Maheshpur (8 years old) than
(12.07), R. reniformis (11.48) and H. erythrinae (11.19) either the younger or older plantations when the diversity
in the community indicating the significance of was based on nematode numbers. The plantation located
prominence and improtance values (Fig. 1) in determining at Rangrung (25 years old) was least diverse in the
the relative ecological role in terms of energy transfer regard (Fig. 2 ). The plantation location at Lakhilunga (20
played by nematode species in a diverse community. The years old) was most diverse when diversity was based on
pathological significance of the M. incognita and M. nematode biomass. This shows that there was a linear
hapla are the most important in tea because it causes increase in the index of diversity of the nematode
severe damage of tea seedlings and younger plants communities with an increase of the age of plantations,
throughout the. world. Pathogenic significance of P. which reaches its highest upto 8 years of age and with
brachyurus comes next to Meloidogyne spp. (Table further increase in the age of plantation, there was a
2). However, the relative pathogenic role played by R. gradual decline in the diversity index in the higher age
renifonnis has been documented from Sri Lanka (Campos groups. This indicated that with an increase in the age of
et al, 1990) which is similar in the present findings. plantation up to certain age, parasitic species have
become more abundant (Table 3). This is a distinct sign
Nematode community diversity of gradual establishment of these species. Beyond certain
age of monoculture, perennial plantation crops do not
Diversity of ptant parasitic nematode community support a very diverse pathogenic community of
was measured by the index of diversity (H’) using phytonematodes. Only the serious and specific plant
nematode numbers (H’n) and biomass (H’b) of individual parasites prevail on a particular crop, as in banana in
species in different tea plantation. These values were West Bengal (Mukherjee & Dasgupta, 1983), in dogwood
correlated with the age of plantations (Fig. 2). Diversity nurseries in Tennesse, USA (Niblack & Bernard, 1985).
index ranged from 1.288 to 1.901 in different nurseries
and plantations when the index was based on nematode Similar findings have been made in the diversity and
numbers. However, diversity index rangedfrom 0.459 to community structure of plant parasitic nematodes in
0.952 when nematode biomass of respective species pineapple plantations (Nath et al., 1997), in rubber
was used (Table 3). It was observed that nematode nurseries and plantations (Mukherjee, et al., 2000) and
co~unity diversity (H’) was less when nematode biomass jackfruit orchards ofTripura (Mukherjee et al., 2001).
of respective species was used instead of numbers. Because plant related inputs provide the resources for
These findings are in agreement with Norton and Edwards nematode communities, soil texture is related to suitability
(1988) in the study of age structure and community of cropping and affects nematode communities through
diversity of nematodes associated with maize in Iowa, crop specific infestations (Yeates, 1999).
USA. Nematode community diversity was the highest in

78
 Table 2: Population density with ranges and frequency of occurrence of plant parasitic nematodes

Population density (Mean ± SE)

NURSERY PLANTATION
Nematode Durga- Harendra- Gokul- Mean Mekhli- Durga- Mahesh- Sabroom Harendra- Mahabir Golak- Lakhil- Gokul- Rangung Mean
species bari nagar nagar para bari pur nagar pur unga nagar

Helicotylenchus 98±20.8 112±77 82±13.2 93±14 150±41 217±45 89±23 264±24.5 174±46.7 378±78 130±44 276±52.5 142±34.6 84±22 182.3±15
spp. (H. dihystera, (10-145) (35-189) (16-156) (53)3 (32-415) (38-450) (22-262) (55-326) (42-430) (24-516) (12-252) (92-382) (25-276) (36-158) (85)3
H. erythrinoe
H. microcephalus)
Hemicriconemoides 10±2.7 28±10.1 - 21±7.2 44±13.5 - 162±84 136±42.4 218±43 82±26 48±18 94±12 26±15.5 102±37 92±13
mongiferoe (6-14) (10-45) (16) (8-115) (78-246) (42-278) (94-360) (35-196) (12-110) (82-106) (4-72) (65-139) (41)
Hoplolaimus spp. 22±3.8 40±10.4 17±10.4 26.1±4.3 14±7.6 45±11 74±19.1 92±14.5 38±8.4 - 30±11 46±11.3 24±2 - 46±5.6
(H. columbus, (8-46) (22-74) (6-38) (60) (5-82) (30-66) (28-120) (10-134) (12-77) (17-128) (25-64) (22-26) (53)

79
H. inducus)
Meloidogyne spp. 15±8.2 98±16.7 135±24 96.1±15 60±10 210±50 113±18.1 178±29.1 280±34.3 47±11.4 124±13.2 142±27.6 38±23.1 55±13 124.3±11.2
(M. hapla (6-48) (35-162) (64-272) (80) (24-110) (82-354) (44-180) (36-220) (92-376) (12-94) (60-175) (86-190) (10-84) (18-76) (70)
M. incognita)
Pratylenchus brochyurus - 27±6.5 48±13 34±6.4 28±4.3 55±8.6 42±9.1 86±15 135±24 44±6 15±4.6 36±11 82±26 20±6.4 53.1±5.1
(12-80) (15-76) (50) (16-60) (20-86) (22-117) (44-158) (62-198) (26-90) (6-32) (25-47) (56-108) (10-32) (68)
Rotylenchulus reniformis 54±9 135±31 86±17.488.5±12.5 - 104±23 217±52.7 358±55.6 50±7.1 156±5.1 34±14 67±15 45±8 37±16.3 53.1±5.1
(28-126) (66-320) (54-235) (93) (72-218) (96-344) (120-482) (35-94) (105-207) (18-160) (52-82) (22-110) (20-86) (55)
Tylenchorhynchus spp. - 16±6.5 28±8.5 22±5.4 - - 32±7.7 - 47±9.1 62±20.5 - - - - 43.6±6.5
(T. brevilineatus, (6-42) (12-55) (33) (8-74) (15-80) (20-115) (22)
T. mashhoodi)
Xiphinema spp. - 16±5.3 23±11 19±5 42±8.1 38±18 60±16.4 - 34±12 26±10 - 54±42 13±4 30±8 37±5
(X. elongatum, (8-26) (12-34) (16) (18-86) (20-56) (38-92) (10-48) (6-52) (12-96) (8-21) (16-44) (28)
X. insigne)
1
Districts (WT = West Tripura, NT= North Tripura, ST = South Tripura, D = Dhalai); 2Age of plantations; 3Frequency of occurrence (%)
Plant parasitic nematodes in tea nurseries and plantations in Tripura
C. Bhattacharya et al.

Table 3: Community analysis of plant parasitic nematodes associated with tea plantations of Tripura.

Nematode Relative Relative Relative Prominence Importance Plantations Pathogenic

species (1) density frequency biomass value value code(3) significance
(%) (%) (%) ranking( 2)

Meloidogyne hapla 8.95 5.86 55.83 528.4 70.64 3-8 2

4
M. incognita 12.07 12.43 36.35 1037.4 60.85 1-10 1
Helicotylenchus dihystera 18.50 14.56 0.21 1720.5 33.27 1-104 -
4
Rotylenchulus reniformis 11.48 9.76 1.53 875.1 22.77 2-10 4
5
Helicotylenchus erythrinae 11.19 7.99 0.10 771.4 19.28 1,2,4,5,8,9 -
Pratylenchus brachyurus 5.25 12.07 1.70 445.2 19.02 1-10 3
Hemicriconemoides magniferae 9.83 7.28 0.14 646.7 17.25 1-104 -
Hoplolaimus indicus 4.38 9.41 0.71 327.6 14.50 1-5,7-9 -
Xiphinema insigne 3.60 4.97 1.25 195.7 9.82 2,3,5,8,9 -
Tylenchorhynchus 4.57 3.90 0.07 220.4 8.54 3,5,6 -
brevilineatus
Helicotylenchus microcephalus 4.28 4.08 0.09 211.0 8.45 3,6,7,10 -
Hoplolaimus columbus 1.65 4.08 1.00 81.5 6.73 1,3,6,8,9 -
Xiphinema elongatum 2.43 2.30 0.86 90.1 5.59 1,6,8,10 -
Tylenchorhynchus mashhoodi 1.75 1.24 0.14 47.6 3.13 3 -

1. Nematodes are arranged in the descending order of their importance values; 2. Based on biological understanding (Rama &
Dasgupta, 1987); 3. Plantations code: (i) Mekhlipara, (ii) Durgabari, (iii) Maheshpur, (iv) Sabroom, (v) Harendranagar, (vi) Mahabir,
(vii) Golokpur, (viii) Lakhilunga, (ix) Gokulnagar, (x) Rangrung; 4. Most widely distributed; 5. Least widely distributed

DISCUSSION Table 4: Nematode community diversity indices based on

nematode number and biomass in tea plantations of
This result is indicated that with an increase in the Tripura, India
age of plantations up to eight years, parasitic species Index based on Index based on
have become more abundant. This is a distinct sign of nematode number nematode biomass
gradual establishment of these species. Diversity of
Age of Index Age of Index
plant parasitic nematode communities was measured by
plantations1 (H’n) plantations (H’b)
the index of diversity (H’) using nematode numbers (Years) (Years)
(H’n) and biomass (H’b) of individual species of diferent
tea plantations. H’n was at its highest in the plantation 08 1.901 20 0.952
located at Maheshpur (8 years of old). H’b was most 10 1.772 22 0.866
diverse at Lakhilunga (20 years of old). 12 1.660 15 0.865
18 1.523 12 0.836
Meloidogyne hapla, M. incognita, Pratylenchus 15 1.522 10 0.812
brachyurus and Rotylenchulus renifonnis have been 05 1.512 18 0.809
recorded in tea nurseries and plantations in Tripura state. 20 1.481 25 0.759
Severe infestations by M. hapla, M. incognita and P. 22 1.402 05 0.537
brachyurus have been encountered throughout the 03 1.388 08 0.475
state and also for the first time from North Eastern states 25 1.288 03 0.459
of India which required immediate attention for nematode 1
Age of plantations arranged by descending diversity of
nematode community

80
 Plant parasitic nematodes in tea nurseries and plantations in Tripura

management. This result is significant from poor Gowen, S.R. & Edmunds, J.E. (1973). An evaluation of some
productivity in tea plantations in Tripura. simple extraction techniques and use of hydrogen
peroxide for estimating nematode population in banana.
Plant Disease Reporter 57: 678-681.
ACKNOWLEDGEMENTS
Jackson, M.L. (1967). Soil chemical analysis Publ. Prentice -
The authors are grateful to Dr. S. Ganguly, Professor, Hall of India. New Delhi, pp 498.
Division of Nematology, IARI, New Delhi and Dr. H.K.
Bajaj, Department of Nematology, HAU, Hissar, Koshy, P.K. (2002). Dr. G,I.D Souza, Memorial Lecture on
Haryana for confirmation of nematode species and Nematodes diseases of plantation crops. Indian Journal
Indian Council of Agricultural Research, New Delhi for of Nematology 32: 107-117.
financing the research scheme and the Head, Department
Mukharjee, B. & Dasgupta, M.K. (1982). Community analysis
of Zoology, M.B.B. College, Agartala for laboratory
of plant parasitic nematodes in tea plantations of West
facilities. C. Bhattacharya is grateful to her Late Bengal; India. Nematologia Mediterranea 10: 1-7.
supervisor Dr. B. Mukherjee for her research work and
Ph.D degree. Mukherjee, B. & Dasgupta, M.K. (1983). Community analysis
of nematodes associated with banana plantations in the
REFERENCES Hoogly district, West Bengal. Nematologia
Mediterranea 11: 43-48.
Anonymous. (2001). Tea Bulletin 2000-2001 Directorate of
Economics and Statistics, Govt. of Tripura, Agartala,1- Mukherjee, B., Nath, R.C. & Dasgupta, M.K. (2000). Plant
30. parasitic nematode communities in rubber nurseries and
plantations in Tripura. Indian Journal of Nematology
Bhattacharjee, P.N. & Singh, R.G. (1995). Tea plantation and 30: 170-174.
the tribes of Tripura Publ. Tripura State Tribal cultural
Research Institute and museum, Govt. of Tripura, pp. 1- Mukharjee, B., Nath, R.C. & Dasgupta, M.K. (2001). Density,
99. Diversity and community Structure of plant parasitic
nematodes in Jackfruit orchards in Tripura. Pest
Campos, V.P., Sivapalan, P. & Gnanapragasm, N.C. (1990). Management in Horticultural Ecosystems 7: 92-97.
Nematode parasites of coffee, cocoa and tea. In: .Plant
parasitic nematodes in subtropical and tropical Nath, R.C., Mukherjee, B., Dasgupta, M.K. & Siddiqi, M.R.
agriculture. Eds, Luc, M. Sikora, R.A & Bridge, J. Publ: (1997). Density, diversity and community structure of
CAB International, UK, pp. 387- 430. plant parasitic nematodes in pineapple plantations of
Tripura, India, International Journal of Nematology 7:
Dasgupta, M.K., Mukherjee, B., Bhattacharya, C., Khan 51-56.
Matiyar, R. & Ghosh, S. (2010). Nematode Infestation
in tea, In: Nematode Infestations Part -II: Industrial Niblack, T.L. & Bernard, E.C. (1985). Nematode community
Crops, Publ : The National Academy of Sciences, India, structure in dogwood, maple and peach nurseries in
213-255. Tennessee. Journal of Nematology 17: 126-13.

Eisenback, J.D. (1985). piagnostic characters useful in the Whitehead, A.G. & Hemming, J.R. (1965). A comparison of
identification of the four most common species of root- some quantitative methods of extracting small vermiform
knot nematodes (Meloidogyne spp.) In : An Advanced nematodes from soil. Annals of Applied Biology 55: 25-
Treatise on Meloidogyne, vol-I : Biology and control, 38 .
Eds, Sasser, J.N. and Carter C.C. North Carolina State
University Releigh, USA, pp. 95-112. Yeates, G.W. (1999). Effects of plants on nematode community
structure. In: Annual Review of phytopathology.
[Link], R.K. Share G & Alfen, N.V.K. 37: 127-149.

81
Mujeebur Rahman Khan et al.
Vol. 43, No. 1, pp. 82-85 Indian Journal of Nematology June, 2013

Role of Salicylic Acid and Phenols in the Resistance of Tagetes species

Against Meloidogyne incognita
MUJEEBUR RAHMAN KHAN*, HINA RIZVI AND RIZWAN ALI ANSARI

Department of Plant Protection, Aligarh Muslim University, 202 002, India

*Corresponding author, Email: mrkhan777in@[Link]

Received on 16-04-2013 and Accepted on 25-05-2013

ABSTRACT: To understand the basis of mechanism of host resistance in marigold spp. against root-knot nematode, Meloidogyne
incognita, marigold, Tagetes erecta cv. Hazara and YD, and T. patula cv. Jafri were grown in pots inoculated with 500, 2500 and 5000
juveniles of M. incognita/kg soil, and the biochemical (chlorophyll, total phenol and salicylic acid), histopathological (root
penetration), and morphological responses of the host plant (length, fresh and dry weight of shoot and root) and soil population
of the nematode were determined. The marigold cultivars were found resistant to root-knot nematode and did not supported gall
formation or egg mass production. Nematode population drastically decreased over time, and only 3-11% larvae could penetrate
the roots, the penetrated juveniles remained confined to cortex and did not attain maturity. Plant growth variables and leaf pigment
were not influenced, but total phenol and salicylic acid contents of leaves of marigold cultivars growing in the infested soil showed
drastic increase (P<0.001) and varied linearly with the inoculum level. The concentration of the biochemicals was relatively greater
in the host cultivar which supported lesser root penetration. This resistance was mainly biochemical rather than morphological.
The small proportion of nematode population that penetrated into roots was not able to grow normally due to synthesis of greater
amount of phenols and salicylic acid. Marigold cultivars not only antagonized the penetrated juveniles, but also suppressed the
nematode larvae in soil and caused mortality to them apparently through root exudates.

Key words: Meloidogyne incognita, phenols, resistance, salicylic acid, Tagetes

Marigold (Tagetes spp.) is one of the important root penetration of M. incognita juveniles in marigold
ornamental plants grown in India and is widely cultivated using different inoculum levels of M. incognita to
in Maharashtra, Gujarat, Andhra Pradesh, Tamil Nadu, understand the mechanism of resistance in marigold
Kerala and Karnataka. Tagetes spp. possesses repellant against the nematode.
and antagonistic character to several pathogens, especially
nematodes (Khan et al., 2012b). Some important MATERIALS AND METHODS
nematoxic compounds such as á-terthienyl have been
found in Tagetes spp. (Meijneke and Oostenbrink, 1957) The experiment was conducted in earthen pots (6 x
which causes toxic effect on plant nematodes 6 inch size) filled with 1 kg autoclaved soil and farmyard
(Devakumar, 1994). Meloidogyne incognita is an manure (4: 1 ratio). Pure culture of second stage juveniles
important pest of ornamental crops (Khan and Mustafa, of root-knot nematode, M. incognita (Kofoid and White)
2005), but most of the Tagetes spp have shown complete Chitwood was prepared from the egg masses collected
resistance to the nematode (Tyler, 1938; Oostenbrink et from the roots of egg plants growing in a pure culture
al., 1957). Ample research has been done to use Tagetes bed. The egg masses were placed on a wire gauge in
spp. as green manuring or intercropping to control root- Petri plates having adequate water and incubated at
knot and other nematodes (Suatmatdji, 1969; Supratoya, 27±2°C for a week to facilitate egg hatching (Khan,
1993; Khan et al., 2012a). The present study is aimed to 2008). The nematode juvenile suspension was collected
examine biochemical and histopathological basis of and standardized to 500 J2 of M. incognita/ml, which
resistance in T. erecta and T. patula with regard to plant was used to inoculate soil in pots to achieve initial
growth, leaf chlorophyll, phenol and salicylic acid and population equivalent to 500, 2500 and 5000 J2/kg soil. A

82
 Salicylic acid and phenols in the resistance of Tagetes species

day after nematode inoculation, 3 week old seedlings of slides to make the surface plain and were examined
T. erecta L. cvs. Hazara and YD and T. patula cv. Jafri under stereoscopic microscope, and the number of
were planted. Ten replicates were maintained for each juveniles and adults of M. incognita inside the root tissue
treatment, and the pot were placed on the roof top were counted.
receiving uniform sunlight in a completely randomized
designs. Pots were watered regularly to maintain adequate Remaining five pots of each treatment were
soil moisture. harvested three months after transplanting and length of
root and shoot, fresh and dry weight of root and shoot, soil
Five out of ten replicates were used to determine population of M. incognita, galls and egg masses were
biochemical and root penetration. Fifteen days after determined. Soil population of juveniles of M. incognita
planting, a few leaves were carefully collected (1 g/ was determined using Cobb’s decanting and sieving
plant) from the 5 plants to determine total phenol (Zieslin method (modified) followed by the Baermann funnel
and Ben-Zaken, 1993) and salicylic acid contents of technique (Southey, 1986). The data were analyzed by
leaves (Shane and Kowblansky, 1968). To estimate single factor ANOVA and least significant difference
chlorophyll (Arnon, 1949), fresh leaves (1 g/plant) were (LSD) was calculated at P < 0.05, 0.01 and 0.001.
collected from the same five plant 15 days later (total
days after transplanting). After collecting leaves, the RESULTS AND DISCUSSIONS
plants on same day, were carefully uprooted from the
pots to determine juvenile penetration in marigold roots Marigold cultivars inoculated with 500, 2500 or 5000
(Southey, 1986). The roots were washed and cut into J2/kg soil did not develop any symptoms that could be
small pieces of 3-5cm. The pieces were plunged into attributed to the nematode damage. Any swelling that
boiling 0.1% acid fuchsine for 3 minutes in a beaker. The could resemble to gall or egg mass production was not
pieces were later washed in running water and placed in noticed on the root system of any of the three marigold
Petri plates with plain lactophenol for 2-3 days allowing cultivars evaluated (Table 1). Soil population of the
differentiation of nematodes from the root tissue. The nematode at the used inoculum levels significantly
small parts of roots were gently pressed between 2 glass decreased over time (P < 0.001), being 59-95% (T.
Table 1. Galling, egg mass production, soil and root population of Meloidogyne incognita in Tagetes species at different
inoculum level of Meloidogyne incognita

Tagetes spp. Inoculation level Total chlorophyll Gall/Egg Soil population of nematodes Root population
(J2/kg soil) (µg/g fresh leaf tissue) mass (J2/kg soil) (larvae/g root at
Months after inoculation harvest)
One Two Three

T. erecta cv. Hazara 500 2.148 00/00 205c 100 c 26 c 2

c c c
T. erecta cv. YD 500 1.644 00/00 925 350 102 4
T. patula cv. Jafri 500 1.66 00/00 925 c 375 c 88 c 7
T. erecta cv. Hazara 2500 2.107 00/00 950 c 375 c 115 c 4
c c c
T. erecta cv. YD 2500 1.59 00/00 1400 600 150 10
T. patula cv. Jafri 2500 1.632 00/00 1450 c 650 c 100 c 13
T. erecta cv. Hazara 5000 2.084 00/00 1500 c 650 c 185 c 8
c c c
T. erecta cv. YD 5000 1.562 00/00 2973 1308 419 14
T. patula cv. Jafri 5000 1.603 00/00 3121 c 1393 c 378 c 13

Each value is mean of five replicates. Significantly different from the respective initial inoculum level) at P < 0.05a, 0.01b, 0.001c

83
Mujeebur Rahman Khan et al.

erecta cv. Hazara), 30-94% (T. erecta cv. YD) and 25-
92% decrease (T. patula cv. Jafri) over respective initial
populations. Histopathology of the roots showed that 3-
11% juveniles of M. incognita penetrated the roots
(Table 1). Highest root population of M. incognita was
recorded in T. patula cv. Jafri (7-11%) followed by T.
erecta cv. Yellow Small (5-9%) and T. erecta cv.
Hazara (3-6%) at 2500 and 5000 J2 level (Table 1). The
root population comprised of only vermiform stage, any
saccate stage or egg masses were not observed.
Fig. 1. Phenol and salicylic acid contents of leaves of Tagetes
Researchers have shown that in resistant plants a small spp. in response to inoculations with Meloidogyne incognita
population of nematodes may penetrate but they do not
develop further to attain maturity (Gaunguly and Gupta,
1994). Inoculation with M. incognita at any inoculum Haq, 2011). The drastic increase in the phenol contents
level did not cause a measurable effect on any of the in response to nematode inoculation in all three cultivars
plant growth and dry matter parameters of T. patula cv. recorded in the present study supports the role of phenols
Jafri, T. erecta cvs. YD or T. erecta cv. Hazara (data on nematode resistance in plants.
not presented). Marigold is an established antagonistic
Salicylic acid contents also got influenced with the
plant for root-knot nematode and most of the Tagetes
nematode inoculation. A significant increase in the salicylic
cultivars are resistant or poor host for this nematode
acid content was recorded with 2500 J2 (Pd”0.01) and
(Tylor, 1938). Apparently, the decrease in population of
5000J 2 (Pd”0.001) of T. erecta cv. Hazara over
M. incognita in the rhizosphere of marigold cultivars
uninoculated control (Fig. 1). In other two cultivars
was due to release of some nematoxic compounds such
increase in salicylic acid due to nematode inoculation
as á-terthienyl along with root exudates (Meijneke and
was significant at P d” 0.05. Salicylic acid is considered
Oostenbrinnk, 1957). Recently Buena et al. (2007)
as a key factor in inducing systemic acquired resistance
screened T. patula against some populations of M.
in plants against pathogens (Ryals et al., 1996; Wobbe et
incognita, M. javanica, M. arenaria and M. hapla
al., 1996). Tolerant or resistant plants synthesize salicylic
and concluded that marigold was resistant to all populations
acid in greater concentrations to avert the pathogen
except race A of M. hapla.
attack (Ryals et al., 1996). Greater concentration of
Inoculation with M. incognita did not influence salicylic acid was recorded in the cv. Hazara which
chlorophyll contents of any of the marigold cultivars supported ingress of juveniles in roots relatively less than
(Table 1). However, total phenol contents of marigold other two cultivars.
cultivars in response to nematode inoculations increased
The present study has demonstrated that marigold
with the increase in inoculum level of the nematode. At
cultivars tested were found resistant to root-knot
500 inoculum level, the phenol contents of leaves increased
nematode, M. incognita and did not developed any
by 24% in T. patula cv. Jafri to 32.5% in T. erecta cv.
galling or egg mass production. This resistance was
Hazara over respective controls (Fig. 1). Same varietal
mainly biochemical rather than morphological. The small
response was recorded at rest of the inoculum levels of
proportion of nematode population (3-11%) that
the nematode and the increase in the phenolic contents
penetrated into roots was not able to grow normally and
was 60-80% and 90-195% with 2500 and 5000 J2/pot,
did not attend maturity evidenced by absence of saccate
respectively. Phenolic compounds are considered
females, galls and egg masses. This has revealed that
important in imparting host resistance and also inducing
although a limited number of larvae could penetrate the
systemic acquired resistance in plants (Kuc, 1995).
roots, but because of synthesis of greater amount of
Greater synthesis of phenol compounds in chickpea and
phenols and salicylic acid the nematode pathogenesis
tobacco plants inoculated with root-knot nematodes has
could not proceed further. Marigold cultivars not only
also been reported earlier (Pankaj et al., 2005; Khan and
84
 Salicylic acid and phenols in the resistance of Tagetes species

antagonized the penetrated juveniles, but also suppressed Meijneke, C.A.R. & Oostenbrink, M. (1958). Tagetes ter
the nematode larvae in soil and caused mortality to them bestriding van aaltjesaantastingen. Meded Dir Tunip
21: 283-290.
apparently through root exudates.
Oostenbrink, M. (1960). Tagetes patula L. als voorvrucht van
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of nematodes infesting horticultural crops. In: Nematode banana. Ilmu Pertanian. 5: 681-691.
infestations Part-III Horticultural Crops. M.R. Khan &
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Khan, Matiyar R., Nagesh, M. & Khan, M.R. (2012b).
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33: 275-297. 339.

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Vol. 43, No. 1, pp. 86-89 Indian Journal of Nematology June, 2013

Efficacy of Essential Oil Extracts of Medicinal Plants Against Rice Root-

Knot Nematode Meloidogyne graminicala in Pots
L. JOYMATI*, N.G. ZENITH , KH. RONIBALA AND P. PURNIMA

Department of Zoology, Nematology Laboratory, D.M. College of Science, Imphal-795001, India

*Corresponding author, E-mail: drjoymatidevi@[Link]

Received on 20-05-2013 and Accepted on 31-05-2013

ABSTRACT: A tub experiment was conducted to study the efficacy of essential oil extracts of medicinal plants against Meliodogyne
graminicola on common rice variety [Link] oil extracts obtained from five selected medicinal plants viz. Parkia javanica,
Zanthoxylum acanthopodium, Jatropha curcas, Vitex negundo and Adatoda vasica were tested as seed soaking against rice root knot
nematode. The seeds were soaked in 100ppm concentration for 24 hours. The plants treated with essential oil extract of Jatropha
curcas showed improvement in plant growth and reduction in diseases incidence when compared with other treated plants. Essential
oil extracts of Zanthoxylum acanthopodium was found to be least effective among five tested plants.

Key words: Essential oil, Meloidogyne graminicola, Oryza sativa( local variety Dharam.)

Rice is believed to have first been domesticated in by this nematode on rice, the present investigation were
India, although the oldest surviving written record telling undertaken to evaluate essential oil extracts of medicinal
of its existence 500 years ago is from [Link] is staple plants against root knot nematode on local breed Dharam
food for about half of the population of our planet and (local rice variety) in tub experiments.
80% or more of the world rice production is reported to
be cultivated in the highly populated southern and eastern MATERIALS AND METHODS
parts of Asia. Rice is grown in almost all the states of
India as a principal food crop. Root knot nematode, Healthy leaves of Parkia javanica, Zanthoxylum
Meloidogyne graminicola is recognized as one of the acanthopodium, Jatropha curcas, Vitex negundo
serious nematode pest of rice that causes root knot and Adatoda vasica were collected, washed with water
diseases characterised by the presence of root galls or and oven dried at 58±2°C for 48 hours before making into
knot on the roots, yellowing, stunting and wilting of the powder with the help of a domestic grinder. For extraction
plant is related to nematode-to-root biomass ratio (Singh of oil 50g dry weight of each plant porduct was taken and
et al., 2006). Nationally M. graminicola is reported to the essential oil extracts was done with the help of
cause upto 50% loss in grain yield (Rao & Biswas, 1973) Clevenger apparatus. The solvent was distilled off and
and in severe cases it may go upto 64% (Phukan, 1995) transferred into a separate beaker, which was completely
Losses in grain yield were also estimated to range from evaporated from the extract in oven till it become semi
16-32 % due to this nematode (Rao & Biswas, 1973). solid material. The essential oil extracts was emulsified
This diseases is widely distributed in rice growing areas with triton X-100 as emulsifier. The seeds of rice plant
of the world. Occurrence of the diseases was reported (Dharam local variety) were soaked in stock solution of
from different states of India by several workers including different oil extract mention above for 24 hours. Then,
Western and Central part of Uttar Pradesh (Kamalwanshi the seeds were spread and allow to dry under shade
et al., 2002 and Pankaj et al., 2010).Considering the condition before sowing. The seeds were soaked in
damaging potential and heavy economic losses caused distilled water served as control. The seed from each

86
 Oil extracts of medicinal plants against rice root-knot nematode in pots

treatment were sown in 30 cm tub containing 5 kg soil. in Jatropha curcas treated plant. The essential oil
Each treatment were replicated three times. After three extract of Zanthoxylum acanthopodium was found to
weeks of sowing, plants were inoculated with freshly be least effective among the plant extracts.
hatched 1000 J2 of Meloidogyne graminicola. The
control was also inoculated and replicated three times. In case of the incidence of diseases, there were
reduction in infestation rate in Jatropha curcas oil
Thirty five days after inoculation of J2 the mature extract treated plant by recording 7 galls and total
plants were uprooted with the help ofhoe and gently polulation of only 580 indicating nearly half times reduction
washed with running tap water. The plants were cut at from initial inoculum level whereas in case of untreated
the margin ofroot and shoot. Length of the root and the one 23 number of galls with l690 nematode population
shoot was measured with the help of meter scale. Fresh were recorded indicating one and half times increased in
weight of the root and the shoot was determined by population level (Table 2). The results supported works
physical balance. Root and shoot were kept separately in of Stirpe et al., 1976 who reported that Jatropha oil cake
paper envelopes and kept in an incubator maintained at is toxic to mammals due to presence of curcin and alectin
58°C for 5 days. Dry weight of root and shoot was which is conformity with the present findings. The seed
determined. The number of grains were also oil of Jatropha is also reported to possess insecticidal,
[Link] of galls, root knot index (0-5 scale) and mollucicidal ,fungicidal and nematicidal properties (Chitra
final nematode population (both in soil and root) were & Dhyani, 2006). Similar results was obtained on reduction
also recorded. of root knot galls and total population with the use of
botanicals as reported by Verma & Khan (2004). Plant
RESULTS AND DISCUSSION treated with the oil extract of Zanthopodium
acanthopodium was found to be least effective with the
The germination percentage of rice seeds was not 16 number galls and 1310 nematode population. From the
affected by seed treatment with oil extract of medicinal above observations, it can be concluded that treated
plant and it was similar and better in all the treatments plants had low infestation rate and improvement in
(Table 1). In general, all the treatment showed better overall growth parameters in comparison to control but
plant growth but also reduced the damage by root knot inoculated one (Table l).
infestation of rice as compared to control. It is clearly
indicated from the data in Table-1 that shoot and root The present investigation is in conformity with those
length of rice plant treated with Jatropha curcas was reported by Kumar et al. (2011) they also reported
maximum followed by rice plant treated with Adhotoda management of root knot and reniform nematode in pots
vasica. The highest number of grains was also recorded and suggested the effectiveness of the oil extracts of

Table 1. Effect of oil extract of medicinal plants against rice root-knot nemoatode infecting rice plant Dharam. in plant growth
parameters

Medicinal Root Shoot Fresh Fresh Dry Dry Number of

Plants Length Length Shoot Root Shoot Root grains
(cm) (cm) wt. (g) wt. (g) wt. (g) wt. (g)

P. javanica 5.96 30.5 8.99 3.01 6.77 1.45 41

X. acanthopodium 5.76 30.46 8.82 2.9 6.50 1.02 39
J. curcas 6.54 33.7 11.9 4.18 9.67 2.86 56
V. negundo 5.52 31 10.5 3.56 8.09 1.07 45
P. thyrsiflorus 5.99 32.1 10.9 3.98 8.87 1.91 48
Control 3.76 29 7.99 2.04 5.77 1.01 35

87
L. Joymati et al.

Table 2. Effect of oil extract of medicinal plants on nematode population against root knot nematode infecting Dharam local rice
variety.

Medicinal Number of Soil pop. Root pop. Root-knot Total Rf

Plants Galls/plant (500 g of soil) /plant Index Pop.

P. javanica 13 580 460 2 1040 1.04

X. acanthopodium 16 880 430 2 1310 1.31
J. curcas 7 360 220 1 580 0.58
V. negundo 10 730 320 2 1110 1.11
P. thyrsiflorus 9 430 390 1 820 0.82
Control 23 920 770 3 1690 1.69

Fig. 1. Effect of oil extract of medicinal plants against rice Fig. 2. Effect of oil extract of medicinal plants on nematode
root-knot nematode infecting rice plant Dharam. in plant population against root-knot nematode infecting Dharam.
growth parameters local rice variety

sacred basil over the other extracts. This study was ACKNOWLEDGEMENTS
supported by Dora and Neog (2006) who tested oil cakes
against root knot nematode larvae in terms of their The authors greatly acknowledged to the D.S.T.
mortality rate. The work also supported the finding of New Delhi, for providing financial assistance and the
Raider & Askary (2011) investigated seed extracts of 17 Principal of the D.M. College of Science, Imphal, for
plant extracts against M. incognita juvenile for their providing laboratory facility during the course of studies.
larval mortality and found inhibitory effects. Rena et al.
(2009) tried out different extracts of plants against M. REFERENCES
incognita and found effective. The above investigation
Bora, B.C. & Neog, P.P. (2006). Effect of oilcakes for management
supported works of Joymati et al., 2012 who reported the
of Meloidogyne iocognita on tea. Annals of Plant
essential oil extract of medicinal plants was found to be Protection Sciences 14: 522-523.
effective against the root knot nematode on kidney bean.
Prasad et al. (2002) reported some plant extracts were Chitra, S. & Dhyani, S.K. (2006). Insect pest of Jatropha
toxic to root-knot nematode and further application in curcas [Link] the potential for their management. Current
pots enhanced plant growth significantly with reduction Science 91: 162-163.
in the nematode population.

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 Oil extracts of medicinal plants against rice root-knot nematode in pots

Joymati, L., Christina, Kh. & Ronibala, Kh. (2012).Effect of sugercane. Annals of Plant Protection Sciences 19:
essential oil extract of medicinal plant against root knot 433-436.
nematode on kidney bean. Annals of Plant Protection
Sciences 20: 441-443. Rao, Y.S. & Biswas, H. (1973). Evaluation of yield losses in
rice due to root-knot nematode, Meloidogyne
Kamalwanshi, R.S., Ganguly, S. & Mishra, S.D. (2002). Indian graminicola. Indian Journal of Nematology 31: 74.
Journal of Nematology 32: 222.
Rena, R.D., Tiwari, M.P., Dwivedi, K., Ansari, N.A. &
Kumar, V., Singh, R.V. & Singh, H.S. (2011). Management of Tiwari, J.P. (2009). Nematicidal effect of angiospermic
Meloidogyne incognita Race 1 and Rotylenchulus medicinal plants against Meloidogyne incognita on
reniformis by seed treatment with bio-agents, organic brinjal. Annals of Plant Protection Sciences 17: 272-
cakes and pesticides on cowpea. Annals of Plant Protection 273.
Sciences 19: 164-167.
Stripe, F., Pession Bazzi, A., Lorenzoni, E., Strochi, P.,
Pankaj, Sharma, H.K., Singh, K. & Lal, J. (2010). Indian Journal Montanaro, L. & Sperti, S. (1976). Studies on the
of Nematology 40: 247-248. proteins from the seeds of Croton tigilium and
Jatropha curcas. Biochemistry Journal 156: 16.
Phukan, P.N. (1995). Nematode problem ofrice crops in India IV.
Stem nematode anb rice root knot nematode In: Nematode Singh, I., Gaur, H.S., Briar, S.S., Sharma, S.K. & Sakhuja,
pest management an appraised of ecofriendly approaches P.K. (2003). International Journal of Nematology
(G. Swarup, D.R. Dasgupta and Gill J.S. eds). Nematological 13: 79-86.
society of India,New- Delhi,India,pp.156-160
Verma, A.C. & Nehal Khan, M. (2004). Potentiality of
Prasad, D., Ram, D. & Ahmad, I. (2002). Managemant of plant botanicals for managing Meloidogyne incognita in
parasitic nematodes by the use of botanicals. Annals of Ocimum canun. Annals of Plant Protection Sciences
Plant Protection Sciences 10: 360-364. 12: 464-465.

Raider, M.G. & Askary, T.R. (2011). Management of plant

parasiric nematode through botanicals and growth of

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Indian Journal of Nematology Vol. 43, No. 1
Vol. 43, No. 1, pp. 90-112 Indian Journal of Nematology June, 2013

SHORT COMMUNICATION

Management of Root-Knot Nematode, Meloidogyne incognita Race-II

(Kofoid and White, 1919) Chitwood, 1949 infesting Brinjal
(Solanum melongena L.)
M.P. ZORE, S.A. PAWAR, N.L. MHASE AND A.G. CHANDELE

Department of Agricultural Entomology, Mahatma Phule Krishi Vidyapeeth, Rahuri-413 722, Dist. Ahmednagar (Maharashtra State), India
*Corresponding author, E-mail: pawarsa2007@[Link]

Received on 08-09-2012 and Accepted on 02-01-2013

Brinjal or egg plant, Solanum melongena (Lin.) 10 kg/ha in combination with ne em cake @ I t/ha,
belonging to family solanaceae, is a native of India. It is carbofuran 3 G @ I kg a.i./ha including an untreated
grown in all South-East Asian countries. Brinjal is control replicated three times at Randomized Block
cultivated throughout the warmer regions of the world Design. The observations at initial and final root-knot
except in the regions having severe winter. Besides, its nematode population, number of root galls and egg
nutritional value, it is well known for its medicinal value masses, gall index/plant and yield at termination were
and is useful against liver complaints, toothache, diabeties recorded. The data obtained were statistically analysed
etc. It is also a good appetizer (Chaudhari, 1977). and presented in Table 1 to 3.

In India, brinjal is being cultivated round in a year in The initial root-knot nematode population in the field
kharif, rabi and summer season with area 6.07 lakh plots was within the range of 480 to 620 J2/200 cm3 of
hectares and production 10.563 million tonnes. In addition soil.
to pest and diseases, plant parasitic nematodes have also
become limiting factor in successful cultivation of brinjal. It could be seen from the data presented in Table 1
Many species of phyto nematodes have been found that all the treatments were found significantly superior
associated with rhizosphere of brinjal. Amongst, the to an untreated control in reducing the root-knot nematode
root-knot nematode, Meloidogyne incognita race-II is population number of root galls/egg masses and gall
considered to be of great economic importance. index and increasing the yield ofbrinjal.

The experiment was conducted during summer, However, the treatment of P. lilacinus at 10 kg/ha
2011 on brinjal (Cv. Ankur) in the field of AICRP on + neem cake at 1 t/ha was found to be most effective in
Nematodes, Department of Agril. Entomology, M.P .K. reducing root-knot nematode population (51.28 %),
V., Rahuri. The seedlings of brinjal grown in nursery number of root galls/egg masses (71.87 %) and gall index
were transplanted in a root-knot infested field at 60 x 45 (37.33 %) and increasing the yield (34.67 %) of brinjal
cm spacing having 4;20 x.2.25 m gross plot size. The with 1 :5.24 ICBR at termination. This was followed by
bioagents, organic amendments and nematicides were the treatment of P. fluorescens at 10 kg/ha + ne em cake
applied at the time of transplanting. There were eleven at 1 t/ha, Trichoderma plus at 10 kg/ha + neem cake at
treatments viz., neem cake @ 2 t/ha; Pseudomonas 1 t/ha and T. viride at 10 kg/ha + neem cake at 1 t/ha
fluorescens, Paecilomyces lilacinus, Trichoderma which were equally effective and on par with each other
plus and Trichoderma viride, @ 20 kg/ha alone and @ in reducing root-knot nematode population (49.28 to

90
 Short Communication

Table 1. Effect of different treatments on root galls/egg masses and gall index of root-knot nematode, M. incognita race-II
infesting brinjal

Treatment Decline in nematode Number of Gall index/ Yield ICBR

population (%) root galls plant (q/ha)
egg masses/
plant at
termination
Soil application of neem cake at 2 t/ha at 32.17 69.74 4.60 175.00 1:1.05
time of transplanting
Soil application of Pseudomonas fluorescens 36.37 61.54 4.06 184.44 1:4.67
at 20 kg/ha at time of transplanting
Soil application of Paecilomyces lilacinus 37.21 58.60 3.86 189.73 1:5.64
at 20 kg/ha at time of transplanting
Soil application of Trichoderma plus 34.55 70.07 4.53 182.01 1:4.23
at 20 kg/ha at time of transplanting
Soil application of Trichoderma viride 35.83 65.94 4.30 179.89 1:3.85
at 20 kg/ha at time of transplanting
Soil application of carbofuran 3G at 45.67 52.87 3.67 192.91 1:11.49
1 kg a.i./ha at time of transplanting
Soil application of Pseudomonas fluorescens 49.28 37.47 3.33 207.41 1:4.64
at 10 kg/ha + neem cake at 1 t/ha
Soil application of Paecilomyces lilacinus 51.28 33.40 3.13 213.76 1:5.24
at 10 kg/ha + neem cake at 1 t/ha
Soil application of Trichoderma plus 48.31 39.67 3.40 201.37 1:4.056
at 10 kg/ha + neem cake at 1 t/ha
Soil application of Trichoderma viride 47.24 47.34 3.53 200.70 1:4.00
at 10 kg/ha + neem cake at 1 t/ha
Untreated control 0.00 118.73 5.00 158.73 -
S.E. ± 0.96 1.66 8.85 4.44 -
CD (P=0.05) 2.83 4.91 0.26 13.12 -

*Figures in parentheses are arcsin transformed values

Market Rates: 1. Brinjal fruits-Rs. 1000/q, 2. Bioagents-Rs. 250/kg, 3. Carbofuran-Rs. 75/kg, 4. Neem cake-Rs. 7.5/kg, 5. Labour
charges- Rs. 500/ha (Soil application)

47.24 % ), number of root galls/ egg masses (68.44 to (2001) on brinjal and Rao (2008) on acid lime for the
60.06 %), gall index (33.33 to 29.33 %) and increase in management of root-knot nematode.
yield (30.67 to 26.44 %) over an untreated control with
1 : 4.64 to 4.00 ICBR. Hence, in the integrated management of root-knot
nematode population, the soil application of P. lilacinus
The effectiveness of soil application of P. lilacinus with neem cake was found most effective for the
+ neem cake for management of root-knot nematode is management of root-knot nematode.
in conformity with that of Cannayane and Rajendra

91
Indian Journal of Nematology Vol. 43, No. 1

REFERENCES Goswami, B.K. & Singh, S. (2004). Pesticide Research Journal

16: 9-12.
Anonymous (2010). National Horticulture Board. Ministry of
Agriculture, Government of India. Mahajan, R. (1982). Indian Journal of Nematology 12: 375-
419.
Cannayane, I. & Rajendran, G. (2001). Current Nematology
12: 51-55. Rao, M.S. (2008). Nematologia Mediterranea 36: 45-50.

Chaudhari, B. (1977). Vegetables, National Book Trust (8th Sonawane, M.L. & Darekar, K.S. (1985). Current Research
Edn.), New Delhi, India. pp. 48.55. Report 1: 15-17.

Chitwood, B.G. (1949). Proceedings of Helminthology Society

Washington 16: 90-104.

Management of Root-Knot Nematode, Meloidogyne incognita, Race-II

Infesting Pomegranate by using Bioinoculants
S.A. PAWAR, N.L. MHASE, D.B. KADAM AND A.G. CHANDELE

Department of Agricultural Entomology, Mahatma Phule Krishi Vidyapeeth, Rahuri-413 722, Dist. Ahmednagar (Maharashtra State), India
*Corresponding author, E-mail: pawarsa2007@[Link]

Received on 08-09-2012 and Accepted on 06-01-2013

Pomegranate (Punica granatum L.), belongs to are notorious for disease complexes involving fungi,
family Punicaceae is one of the important fruit crops of bacteria, virus, mycoplasma, insects and other nematodes
tropical and subtropical regions. This ancient world fruit (Dasgupta & Gaur, 1986).
originated in Persia, Iran, Afghanistan and Baluchistan
(de Candolle, 1967). The experiment was conducted continuously for
three years during kharif , 2008-09 to 2010-11 at Central
Extensive cultivation of pomegranate has been Campus, Mahatma Phule Krishi Vidyapeeth, Rahuri on
reported in Mediterranerul countries like Spain, Egypt pomegranate (cv. Mrudula) planted at 4.5 x 3 m spacing.
and Morocco. It is also grown to some extent in Burma, The bioagents and nematicides were applied in the
China, Japan, Russia, California (USA) and India. In rhizosphere soil by ring method at ‘Bahar’ (Bloosom).
India, it is grown over an area of 1.13 lakh hectares with There were five treatments viz., Pseudomonas
an annual production of about 7.92 lakh metric tonnes. fluorescens and Trichoderma viride at 20 kg/ha, cartap
hydrochloride (4 G) at 75 kg/ha and carbofuran (3 G) at
Such an important fruit crop is attacked by several 100 kg/ha including an untreated control replicated four
insect and non insect pests as well as diseases. Diseases times at Randomized Block Design. The observations at
caused by nematodes are of economic importance. initial, intermediate and termination on root-knot nematode
Darkekar et at. (1990) reported ten species of plant population and number of root galls/5 g roots and yield/
parasitic nematodes associated with this crop in the tree were recorded. The pooled data obtained were
Maharashtra state. The root-knot nematode, statistically analysed and presented in Table 1.
Metoidogyne incognita is one of the them causing
considerable yield losses in pomegranate. Besides, the The average initial root-knot nematode population in
direct damage caused to the plant, the root-knot nematodes the field ranged from 360 to 600; 400 to 620 and 480 to

92
 Short Communication

Table 1. Effect of different treatments on population of root-knot nematode, M. incognita race-II on pomegranate

Treatment Decline in Root galls/ Yield B:C Ratio

population (%) 5g roots (%) (t/ha)

P. fluorescens at 20 g/m2 31.2 29.2 18.4 1:2.37

T. viride at 20 g/m2 28.3 23.9 18.0 1:2.33
Cartap hydrochloride 27.2 20.4 17.6 1:2.27
4 G at 0.3 g a.i./m2
Carbofuran 3 G at 29.8 25.1 17.8 1:2.27
0.3 g a.i./m2
Untreated control - - 15.4 1:2.27
S.E.± 1.03 0.80 0.36 -
CD (P=0.05) 2.99 2.30 1.17 -

640 12/200 cm3 of soil during 2008-09, 2009-10 and found to be effective for the dontrol of root-knot nematode
2010-11, respectively. by Goswami & Singh (2004). As regards the additional
yield and cost: benefit ratio, the treatment of P.
It could be seen from the pooled data presented in fluorescens and T. viride is in conformity with that of
Table 1 that all the treatments were significantly superior reported by Tadavi (2008). Thus, effectiveness of P.
to an untreated control in reducing the root- knot nematode fluorescnes and T. viride by lowering down the nematode
population and number of root galls/5 g root and increasing population is extending the scope of biological control o
the yield of pomegranate. phytonematodes .

However, the treatment of P. fluorescens at 20 kg/ REFERENCES

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by using bioinoculants. Research Review Committee
pomegranate (18.99 %) with 1:2.37 B:C ratio. It was
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carbofuran (3 G) at 100 kg/ha. The reduction in root-knot M.P.K.V., Rahuri. pp. 32-39.
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of root-knot nematode. This is in conformity with that of Dasgupta, D.R. & Guar, H.S. (1986). The root-knot nematodes,
reported by Hashim (1983), Siddiqui and Khan (1986), Meloidogyne spp. in India. In: Plant Parasitic
Darekar et al. (1989) in pomegranate. Nematodes of India. Problem and Progress [Swarup G.
and Dasgupta, D.R. (eds.)] pp. 139-178.
The results of the present investigation of
de Candolle, A. (1967). ‘Origin of cultivated plants’ Hafner
effectiveness of P. fluorescens is in collaboration with Pub. Co. New York and London. pp. 237-240 and 441.
those reported by Anonymous (2008). T. viride was also

93
Indian Journal of Nematology Vol. 43, No. 1

Goswami, B.K. & Singh, S. (2004 ). Pesticide Research Journal Tadvi, K.M. (2008). Management of root-nematode, M.
16: 9-12. incognita infesting pomegranate. A part of thesis
submitted to Mahatma Phule Krishi Vidyapeeth, Rahuri
Hashim, Z. (1983). Nematologia Mediterranea 11: 199-200. for the fulfillment of [Link]. (Agri.) degree.

Siddiqui, Z.A. & Khan, M.W. (1986). Pakistan Journal of

Nematology 4: 83-90.

Induction of Defence Enzymes in Tomato by Plant Growth Promoting

Rhizobacterium, Pseudomonas fluorescens against Root-Knot Nematode,
Meloidogyne incognita
P.G. KAVITHA*, E.I. JONATHAN AND K. SANKARI MEENA

Department of Nematology, Tamil Nadu Agricultural University, Coimbatore- 641 005, India
*Corresponding author, E-mail: kavitha_nem@ [Link]

Received on 03-10-2012, Resubmitted on 05-02-2013 and Accepted on 15-01-2013

Tomato (Lycopersicon esculentum Mill.) is one of chitinases, glucanases, peroxidase, polyphenol oxidase,
the major vegetable crops grown in India. Root-knot phenylalanine ammonia-lyase and synthesis of
nematodes (Meloidogyne spp.) are major pathogens of phytoalexins are associated with resistance due to
tomato throughout the world, affecting both the quantity application of P. fluorescens.
and quality of marketable yields. Success in nematode
management over a long term requires several Peroxidases are involved in the regulation of plant
alternatives and combinations of approaches to be cell elongation, phenol oxidation, IAA oxidation, oxidation
economically feasible. Currently both naturally occurring of hydroxy cinnamyl alcohols into free radical
and applied biological controls are being utilized to control intermediates and wound healing. Polyphenol oxidase
plant parasitic nematodes. Biological control of nematodes accumulation takes place upon wounding by pathogens
has long been considered as an alternative to managing in plants. PPO is induced via. octadecanoid defense
nematodes with pesticides. signal pathway which is usually associated with feeding
by insects or similar physical trauma. It involves jasmonic
Plant growth promoting rhizobacteria Pseudomonas acid as an intermediate signal and culminates in the
fluorescens which live in close proximity of plant roots, production of proteins such as PPO and proteinase
help in boosting the plant growth and vigor by different inhibitors (Schaller & Ryan, 1995). Phenylalanine
mechanisms are deleterious to the plant pathogens. The ammonia lyase (PAL) catalyzes the deamination of L-
bacteria achieve this mainly by bio-stimulation, bio- phenylalanine to trans-cinnamic acid, which is the first
control, bio-fertilization and bio-remediation. step in the biosysthesis of large group of plant products
such as lignin monomers and phytoalexins. Induction of
Induced systemic resistance (ISR) of plants against PAL, peroxidase and the accumulation of such phenolics
pathogens is a widespread phenomenon that has been occur in response to pathogen attack. Therefore, a study
intensively investigated with respect to the underlying was conducted to assess the induction of the defense
signaling pathways as well as to its potential use in plant enzymes by applying talc-formulation of P. fluorescens
protection. Changes in cell wall composition, de novo against root-knot nematode in to tomato.
production of pathogenesis - related-proteins such as

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 Short Communication

A field experiment was conducted in Coimbatore, The activity of defense enzymes were analyzed in
India on tomato [Link]-1 infested with M. the leaf samples 30 days after application of P.
incognita the talc-based formulations of promising P. fluorescens. Peroxidase activity was assayed
fluorescens isolates were tested against M. incognita spectrophotometrically (Hartee & Keilin, 1955). The
infesting tomato. The talc-based formulation of the reaction mixture consisted of 1.5 ml of 0.05 M pyrogallol,
promising P. flourescens isolates Pft 18, Pft 20 and Pft 0.5 ml of the enzyme extract and 0.5 ml of 1 percent
25, and standard Pf 1 were applied to the soil at two H2O2. The reaction mixture was incubated at room
doses viz., 2.5 g/pot and 3.0 g/pot and chemical carbofuran temperature (28 ± 1 ° C). The change in absorbance was
at 1 g a.i./pot. Untreated control was also maintained recorded at 30 sec interval for 3 min. The boiled enzyme
(Table 1). The plot size of 10 m2 was maintained for all preparation served as blank. The enzyme activity was
the treatments. The formulation was applied 30 days expressed as change in the absorbance at 420 hM/min/
after planting of the tomato seedlings of cv. PKM-1. The g on fresh weight basis (Hammerschmidt et al., 1982).
population densities of P. fluorescens in the formulations Polyphenol oxidase activity was determined as per the
were 2.5 x 10 8 cfu/g. The study was undertaken with the procedure given by Mayer et al., (1965). One g of leaf
following treatments by using randomized design with was used for polyphenol oxidase estimation. The reaction
ten treatments each replicated three times. The treatments mixture consisted of 1.5 ml of 0.1 M sodium phosphate
are as follow T1: Pft 18 @ 2.5 kg / ha; T2: Pft 18 @3.0 buffer (pH 6.5) and 200 ml of the enzyme extract. To
kg / ha; T3: Pft 20 @ 2.5 kg / ha; T4: Pft 20 @ 3.0 kg / start the reaction, 200 ml of 0.01 M catechol was added
ha; T5: Pft 25 @ 2.5 kg / ha; T6: Pft 25 @ 3.0 kg / ha; and the activity was expressed as changes in absorbance
T7: Pf 1 @ 2.5 kg / ha; T8: Pf 1 @ 3.0 kg / ha; T9: at 495 hM/min/g fresh weight of tissue. The assay
Carbofuran @ 1kg a.i./ ha; T10: Control mixture containing 100 ml of enzyme, 500 ml of 50 mM
Tris HCl (pH 8.8) and 600 ml of mM L -phenylalanine

Table 1. Effect of Pseudomonas fluorescens isolates on peroxidase, polyphenol oxidase and phenylalanine ammonia lyase
activities in leaves of tomato cv. PKM-1

Treatment Peroxidase* Per cent Poly phenol Per cent Phenylalanine Per cent
increase oxidase** increase ammonialyase*** increase
over control over control over control
Pft 18 (2.5 kg/ha) 0.45 67.1 0.07 16.9 0.31 7.5
Pft 18 (3.0 kg/ha) 0.42 54.3 0.06 4.6 0.31 6.5
Pft 20 (2.5 kg/ha) 0.77 181.0 0.09 27.6 0.44 52.2
Pft 20 (3.0 kg/ha) 0.76 179.9 0.09 24.6 0.44 51.2
Pft 25 (2.5 kg/ha) 0.32 17.5 0.07 12.3 0.29 34.0
Pft 25 (3.0 kg/ha) 0.31 13.8 0.07 9.2 0.28 32.9
Pf 1 (2.5 kg/ha) 0.68 151.4 0.07 21.5 0.43 49.1
Pf 1 (3.0 kg/ha) 0.57 108.0 0.07 16.9 0.40 38.1
Carbofuran 3G 0.37 36.8 0.07 10.7 0.31 7.9
(1 kg a.i./ha)
Control 0.27 - 0.06 - 0.25 -
CD (P=0.05) 0.17 - 0.01 - 0.05 -

*Changes in the absorbance/min/g leaf tissue at 480 nm, ** Changes in the absorbance/min/g leaf tissue at 495 nm, *** Changes
in the absorbance/min/g leaf tissue at 290 nm

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Indian Journal of Nematology Vol. 43, No. 1

ammonia lyase was incubated for 60 min. The reaction PAL plays a significant role in regulation and accumulation
was arrested by adding 2 N HCl. Later 1.5 ml of toluene of phenolics, which directly related to the degree of plant
fraction containing trans – cinnamic acid was separated. resistance to pathogens. Increased phenolic synthesis is
The toluene phase was measured against the blank of observed in the P. fluorescens treated plants (Graham
toluene. Standard curve was drawn with graded amounts & Graham, 1991).Induction of systemic resistance was
of cinnamic acid in toluene as described earlier. The correlated with the accumulation of defense enzymes
enzyme activity was expressed as hM of cinnamic acid PO, PPO and PAL upon treatment with plant growth
at 290 hM min –1 g–1 of fresh tissue. promoting rhizobacteria, P. fluorescens and there by
reduced the infection by root-knot nematode.
The results showed an enhanced activity of defense
enzymes in tomato plants treated with P. fluorescens. REFERENCES
The Pft 20 @ 2.5 kg/ha treated tomato leaves recorded
0.770 followed by Pft 20 @ 3.0kg/ha which recorded Ganguly, A.K. & Dasgupta, D.R. (1979). Indian Journal of
0.767 and the control being 0.274. A significant increase Nematology 10: 151-189.
in polyphenol oxidase activity was observed in the Pft 20
Graham, T.L. & Graham, M.G. (1991). Molecular Plant
treated plants followed by the application of Pf1, Pft 18 Disease 4: 415 -720.
and Carbofuran. Resistant plants of tomato when
inoculated with nematode M. incognita showed higher Hammerschmidt, R., Nuckles, E.M., & Kuc, J. (1982).
concentration of peroxidase (Zacheo et al., 1982). Physiology and Plant Pathology 20: 73-82.
Sujatha and Mehta (1998) found that there was an
increase in enzyme levels of peroxidase and polyphenol Jonathan, E.I., Barker, K.R., Abdel Alim, F.F., Vrain, T.C., &
Dickson, D.W. (2000). Nematropica 30: 231-240.
oxidase in sugarcane roots infested with M. javanica.
Galled roots contained twice the amount of simple sugars Rodriguez-Kabana, R., Jordan, J.W., & Hollis, J.P. (1965).
and insulin than that of the healthy roots (Epstein, 1972). Science 148: 524-526.
Quantitative increase in peroxidase activities was obvious
throughout the period of observation in the resistant Schaller, A. & Ryan, C.A. (1995). Bioassays 18: 27-33.
tomato in the susceptible tomato plants (Ganguly and
Dasgupta, 1979). Sujatha, K. & Mehta, U.K. (1998). International Journal of
Nematology 8: 102-104.
Phenylalanine ammonia lyase activity significantly
Zacheo, G., Bleve-Zaccheo, T., & Lamberti, F. (1982).
increased in the Pft 20 treated plants by 0.443 when Nematologia Mediterranea 10: 75-80.
compared to the untreated control which recorded 0.251.

Quantitative Estimation of Free Amino Acids and Amides in Okra Inoculated

with Meloidogyne incognita
S. MOHANTA AND K.C. MOHANTY

Department of Nematology, Orissa University of Africulture & Technology, Bhubaneswar- 751 003, India
*Corresponding author, E-mail: swatilekha.agrico5@ [Link]

Received on 05-10-2012 and Accepted on 20-01-2013

Stock cultures of the root-knot nematode, obtained from a single egg mass progeny, maintained and
Meloidogyne incognita for the present study was multiplied on susceptible brinjal variety, Pusa Purple

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 Short Communication

Long. Earthen pots of 15 cm dia were surface sterilized decanted into separate watch glasses and allowed to
with formaldehyde solution (1.0 %) and filled with evaporate till a slight yellowish colour syrupy material
autoclaved soil @ 1 kg/pot. Surface sterilized seeds of obtained. This syrupy material was dissolved in 1 ml of
okra variety L.B.H-55 were planted in pots. One month 10 % isopropyl alcohol and transferred to separate glass
after germination second stage juveniles of M. incognita vials representing for a single lot and stored in the freezer
were inoculated @ 1000 J2 ± 20 per kg soil. Side by side compartment of refrigerator. The above solution was
a healthy uninoculated check was also maintained. After used for estimation of amino acids and amides
45 days of nematode inoculation the plants were uprooted quantitatively by paper chromatographic technique and
and the roots were thoroughly washed with tap water subsequently by using TLC scanner (densitometer).
and double distilled water. Then the roots were surface After measuring the density of various amino acids and
sterilized in 0.1% Mercuric chloride solution followed by amides from developed chromatograms, their
repeated washing with distilled water and taken for concentrations were calculated with the help of standard
analysis of amino acids and amides. Root samples of both graph of known concentration. The results were expressed
healthy and inoculated plants were taken, ground in terms of milligrams of amino acids and amides per
separately in 80 percent ethyl alcohol, boiled for 5 to10 gram fresh roots.
minutes in a hot water bath and filtered through a double
layered fine cheese cloth. The extracts were centrifuged Results presented in Table-1 revealed 9 amino acids
at 5000 rpm for 10 minutes. The supernatants carefully viz. L-cystine, L-arginine, L- serine, L-glutamic acid, L-

Table 1. The Rf value of ninhydrin positive spots on chromatogram of inoculated and un-inoculated okra variety L.B.H-55

Rf. Value of Amino acids Rf. Value of Amino acids

inoculated cultivars and amides un-inoculated and amides
identified cultivars identified

1.4/22.5=0.06xI00=6 L-cystine 1.4/22.5=0.06xI00=6 L-cystine

2.9/22.5=0. 13x100=13 L-arginine 2.9/22.5=0.13xI00=13 L-arginine
4.1/22.5=0.18xI00=18 L-serine 4.1/22.5=0.18xI00=18 L-serine
5.3/22.5=0.24xI00=24 L-glutamic acid 5.3/22.5=0.24xI00=24 L-glutamic acid
2.6/22.5=0. 12x100=12 L-aspargine* 2.6/22.5=0. 12x100=12 L-aspargine*
7.4/22.5=0.33xI00=33 L-alanine 4.8/22.5=0.21xI00=21 L-glycine
2.3/22.5=0. 10x100=10 L-Iysine 7.4/22.5=0.33xI00=33 L-alanine
9.5/22.5=0.42x100=42 L-tyrosine 9.5/22.5=0.42xI00=42 L-tyrosine
8.2/22.5=0.36xI00=36 L-proline 3.7/22.5=0.16x100=16 L-glutamine*
11.5/22.5=0.5IxI00=51 L-tryptophan 2.1/22.5=0.09xI00=9 L-histidine
14.4/22.5=0.64xI00=64 L-valine 11.5/22.5=0.51xI00=51 L-tryptophan
13.7/22.5=0.61xI00=61 L-phenyl alanine 13.7/22.5=0.61x100=61 L-phenyl alanine
16.3/22.5=0. 72x1 00=72 L-Ieucine
17. 1/22.5=0.76xI00=76 L-isoleucine
4.3/22.5=0. 19x100=19 L-aspartic acid
6.0/22.5=0.27xI00=27 L-threonine

* amide

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Indian Journal of Nematology Vol. 43, No. 1

Table 2. Quantity of amino acids and amides in healthy and amino acids in infected plant may be due to progressive
M. incognita inoculated okra (var- L.B.H-55) root hydrolysis of plant proteins by the proteolytic enzymes
secreted by root-knot nematode, M. incognita. The
Name of Healthy Inoculated % increase(+)
amino acids mg/g mg/g or decrease(-) reduction of L- tryptophan (47.83%) in nematode
and amides over healthy inoculated samples might be attributed to the diversion
and synthesis of IAA for gall formation as reported by
L-cystine 0.052 0.069 + 32.69 Krusberg, 1961. Accumulation of L-proline in nematode
L-arginine 0.098 0.087 -11.22 inoculated roots as observed in the present investigation
L-serine 0.049 0.056 + 14.29 is in agreement with Lewis and McClure, 1975; Mohanty
and Das, 1976; Mishra and Mohanty, 2006; Tripathy and
L-glutamic acid 0.075 0.089 + 18.67
Mohanty, 2008. Proline accumulation may be attributed
L-aspargine 0.038 0.048 + 26.31 as a mechanism in plants to overcome nematode induced
L-alanine 0.073 0.091 + 24.66 stress as reported by Epstein, 1972. Further the reduction
of L-arginine in nematode inoculated sample may be due
L-phenyl alanine 0.072 0.089 + 23.61
to conversion of L-arginine to L-proline through Ornithine
L-tyrosine 0.061 0.049 -19.67 cycle. Decrease in aromatic amino acid L-tyrosine in
L-tryptophan 0.046 0.024 -47.83 nematode infected samples in the present study have
significant role in resistance. It is also interesting to note
L-glycine 0.058 - -
that during post infection period some of the amino acids
L-glutamine 0.044 - - appeared and some other disappeared along with an
L-histidine 0.029 - - increase in certain amino acid accompanied by
concomitant decrease in another amino acid as reported
L-aspartic acid 0.056 - -
by Epstein and Cohn, 1971. Such phenomenon suggests
L-Ieucine 0.047 - - possible interconversion of one amino acid from another.
L-isoleucine 0.054 - -
REFERENCES
L-threonine 0.051 - -
L-Iysine 0.056 - - Epsteio, E. (1972). Nematologica 18: 555-562.
L-proline 0.041 - -
Epsteio, E. & Coho, E. (1971). Journal of Nematology 3: 334-
L-valine 0.082 - - 340.

Krusberg, L.R. ( 1961 ). Nematologica 6: 185.

aspargine, L-alanine, L-phenyl alanine, L-tyrosine and
L-tryptophan common to both healthy and inoculated Lewis & [Link], M.A. (1975). Journal of Nematology 7: 10-
roots. However, 7 amino acids specific to healthy roots 15.
namely L-glycine, L-glutamine, L-histidine, L-aspartic
acid, L-Ieucine, L-isoleucine and L- threonine were Mishra, C.D. & Mohanty, K.C. (2007). Indian Journal of
Nematology 37: 131-134.
identified. Similarly amino acids specific to inoculated
root were L-Iysine, L- proline and L-valine. Perusal of Mohanty, K.C., Dash, L. & Swain, S.C. (2001). Annals of Plant
data of both qualitative and quantitative study (Table-2) Protection Sciences 9: 81-86.
indicated increase in most of the amino acids and amide
common to healthy and inoculated roots after post Mohanty, K.C. & Das, S.N. (1976). Indian Phytopathology
infection period except L-arginine, L-tryptophan and L- 29: 434-436.
tyrosine. Similar increase in various amino acids was
Tripathy, M.K. and Mohanty, K.C. (2008). Journal of Research
also recorded by Mohanty and Pradhan, 1990; Mohanty
Orissa university of Agriculture and Technology 26:
et al, 2001; Mishra and Mohanty, 2007; Tripathy and 67-69.
Mohanty, 2008 in different plants. The increase in various
98
 Short Communication

Distribution of Root-Knot Nematode in Ornamentals of Jorhat district,

Assam
KALYAN KR. DUTTA, BINA B. GOGOI AND BHABESH BHAGAWATI

Department of Nematology, Assam Agriculture University, Jorhat- 785013

*Corresponding author, E-mail:

Received on 30-11-2012 and Accepted on 20-01-2013

The ornamental plants and flowers are high valued ornamental gardens, parks, farms and from green houses,
cash crops when grown commercially. The income per distributed randomly throughout the district. A total of
unit area from floriculture is much higher than any other 174 Soil and root samples were collected from the
agriculture products. The economic importance of rhizosphere of different ornamental plants up to a depth
ornamentals has been increasing day by day and so is the of 15-30 cm depending on the depth of the roots. Each
international demand and market. The recent statistics bulk sample was constituted of several sub samples and
shows that the area under cut flowers in our country is portion of roots collected randomly from different sites
64,768 ha (Dadlani, 1999). Five hundred hectare of each locality. Samples were collected with the help of
climatically controlled greenhouses are available for a khurpi or a hoe from the rhizosphere of the crop and
growing quality flowers for export. Though the production then kept in polythene bags and tightly tied with a twine
of ornamental is increasing day by day still the total yield thread. An aluminium tag containing information like
is significantly less as compared to the demand. One the host crop, locality, date of collection was tied with the
main causes is the pest and diseases. Pest attack not only polythene bag carrying the soil and root samples. The
reduces the production of ornamentals quantitatively, but samples were then taken to the laboratory and stored in
also deteriorate the quality of produced thereby reducing refrigerator at about 10º C till the extraction of nematodes
the market value drastically. Among the major constrains was made.
in flower production, plant parasitic nematodes are found
to have deleterious affect on quantitative yield and Concentrated nematode suspension was taken in a
quality of flowers. beaker and left undisturbed for few minutes allowing the
nematodes to settle down at the bottom. An equal
Sasser and Freckman (1987) reported 11 per cent amount of double strength boiling formalin solution (8%)
losses in ornamental plantations throughout the world was added to the suspension in the beaker. The nematode
due to plant parasitic nematodes. Nematodes are reported suspensions were stored in small glass bottle with proper
to be major constraint in ornamental production by label. The root samples were taken out from the polythene
various workers in India (Jayaraman et al. 1975, Khan bags to observe for the presence or absence of galls and
& Pal, 2001). eggmasses on the roots. For staining of nematodes within
the plant tissue, Acid Fuchsin in lactophenol method was
Among the plant parasitic nematodes, root knot followed (Byrd et al. 1983). Cut roots were then wrapped
nematode, Meloidogyne incognita, is a major nematode with a muslin cloth and tied with a twine thread. The
pest infesting almost all the ornamentals. muslin cloth was then plunged into boiling lactophenol
containing 0.05-0.1 per cent acid fuchsin stain and boiled
To study the occurrence and distribution of root-knot for 2-5 minutes.
nematode in ornamental plants of Jorhat district, a random
roving survey was carried out during 2009-2011, covering Preparation of perineal pattern was done for the
seven blocks of Jorhat district. Soil and root samples confirmation of the root-knot species (Taylor and Nester
were collected from different nurseries, homestead 1974).The species was confirmed to be Meloidogyne

99
Indian Journal of Nematology Vol. 43, No. 1

incognita. Dorsal arch was high, squared-off and the occurrence in Baghchung, Dhekorgorah, Majuli, Ujoni
dorsal arch composed of smooth to wavy striae. Some Majuli, Kaliapni, Titabar and Chipahikhola are 56.75,
striae forked near the lateral lines and lateral incisures 64.70, 66.66, 63.63, 70.00, 48.38 and 76.92 per cent
are not so distinct. respectively. The maximum frequency was recorded in
Chipahikhola block i.e. 76.92 per cent, whereas the
A Green house, growing gladiolus, observations minimum frequency of occurrence was recorded in
revealed that, the extent of damage caused by this Titabar block i.e. 48.38 per cent.
nematode was much higher within the greenhouse as
compared to the outdoor plants. Symptoms of infestation REFERENCES
was observed in yellowish / brownish patches. Above
the ground, the plants were stunted and discoloured, Dadlani, W.K. (1999). Prospects of floriculture industry in
nematodes also affected the size and leaf count of the India. In: Floriculture and Landscaping, ed. Bose,
T.K.; Maiti, R.G.; Dhua, R.S. and Das, P.; Naya Prakash,
plants. Culcutta, India, p. 757.

A total of 174 samples were collected randomly Jayaraman, V., Rajendran, G. & Muthu Krishnan, T.S. (1975).
from different blocks of Jorhat district, viz., Baghchung Indian Journal of Nematology 5: 101-102.
(A.A.U Campus and Jamuguri), Dhekorgorah (Namdeori
and Rajpriom nursery), Majuli (Kamalabari), Ujoni Khan, M.R. & Pal, A.K. (2001). Pest Management of Economic
Majuli (Maz Deori gaon), Kaliapni (Teok), Titabar Zoology 6: 27-30.
(Bebejia and Central Muga Research Station) and
Sasser, J.N. & Freckman, D.W. (1987). A World Perspective
Chipahikhola (Kakojan). Root-knot nematode infestation on Nematology. The Role of the Society, In: Verch, J.A.
was recorded in most of the root and soil samples. Out and Dickson, N.W. (Eds.) Vistas on Nematology. Society
of 174 samples, root knot nematode was detected in 103 of Nematologists, Hyattsville, Maryland US, pp. 7-14.
samples from different places. The frequency of

Spectral Signature of Mungbean Infested with Root-Knot Nematode,

Meloidogyne incognita under Different Management Options
CHANDRAMANI WAGHMARE, H.K. SHARMA, PANKAJ, RAVI SAHU* AND JAGAN LAL

Division of Nematology, * Agricultural Physics, Indian Agricultural Research Institute, New Delhi-110 012, India
*Corresponding author, E-mail: hksharma@[Link]

Received on 10-01-2013 and Accepted on 22-02-2013

Spectral reflectance from the plant canopy vary with the mungbean plant canopy infested with different level
conditions which actually are govern by different biotic of root knot nematode Meloidogyne incognita. Does
and abiotic factors however these are specific to it discriminate as per the infestation load. Also an
conditions. These could be part of remote sensing attempt has been made to get the reflectance pattern
exercise to get the ground truth. It has been seen that from the mungbean canopy when the management
chlorophyll content, water and mineral content and overall options are implied against root knot nematode. This can
turbidity of system guide reflectance. In this backdrop form a diagnostic tool for nematode problem identification
an effort was made to get the reflectance pattern from at field situation.

100
 Short Communication

Experiment was conducted in green house condition however at higher wavelength 700-1000um lowest 0.35
in 6" earthen pots filled with 1 kg of steam sterilized soil. and median 0.45 reflectance is observed with low and
Mungbean cv Pusa vishal was used for the experiments. high conc of carbofuran respectively. High reflectance
Experiment was done with six replication in completely of 0.5 and low reflectance of 0.4 has been observed with
randomize design. Root-knot nematode culture was T. harzianum high and low conc. respectively. While
isolated from the brinjal seedling and the inoculation was with neem oil reflectance is similar 0.45 at both the conc.
made as 0. 10, 100,1000, 4000, 6000 J2 per pot (T1, Highest reflectance of 0.6 has been observed with
T2,T3,T4,T5,T6,T7) along with untreated (Fig. 1). untreated un inoculated. Gausman et al ( 1975) observed
Reluctance was captured after 25 days of inoculation . differences between light reflectance from leaves of
In another experiment root knot nematode was managed cotton (Gossypium hirsutum) plants grown with a low-
with carbofuran 1 and 2 kg a.i/ha (T1 and T2), or no-nematode (Rotylenchulus reniformis) population
Trichoderma harzianum 1 and 2 %w/w (T3 and 4) (nonstressed), and from leaves grown with a high
beside neem oil coating of seed at 1 ad 2% w/v (T5 and nematode population (stressed), over the 0.5- to 2.5-/
6) of mungbean along with nematode control (T7) and microm waveband, stressed leaves had lower reflectance
untreated (T 8) (Fig. 2). The reflectance was captured than nonstressed leaves of the same chronological age
after 25 days of inoculation. Reflectance was measured for both field- and greenhouse-grown plants. Reflectance
spectrophotometrically on plant canopies differences between stressed and nonstressed leaves in
the visible (0.5 to 0.75 microm), near-infrared (0.75 to
As shown in figure 1 that reflectance varies with 1.35 mum) and infrared water absorption (1.35 to 2.5
inoculums level. Reflectance has been high (0.3-0.4) microm) regions were primarily caused by differences in
with highest inoculums level 6000 J2/pot in the wavelength leaf chlorophyll concentration, mesophyll structure, and
of 300-700um and lowest reflectance has been with no water content, respectively. Similarly Palacharia et al.
or 100 J2/pot. However, this trend become opposite at (2011) used hyperspectral data for reniform nematode
high wavelength of 700-1000. problem detection in cotton fields. Likewise use of
remote sensing in detection of soil nematode is highlighted
Reflectance from the plant canopy of mungbean in by Chrishtian Hillnhutter et al. (2010). Lawrence, et al.
response to nematode management indicated (fig 2) that (2007) reported nematode management using remote
at lower wavelength 300-700um reflectance are mix one sensing technology, self-organized maps and variable
and could be determined easily which has high or low rate nematicide applications. Nutter et al. (2002) studied

Reflectance Reflectance
1.00 0.70
0.90 T1
0.60
0.80 T2
T1 (control, with Rhi.)
0.70 0.50 T3
T2 (100J2)
0.60 T4
T3 (1000) 0.40
0.50 T5
T4 (2000) 0.30
0.40 T6
T5 (4000)
0.30 0.20 T7
T6 (6000)
0.20 T8
T7 (without Rhi.) 0.10
0.10 T9
0.00 0.00 T10
300 400 500 600 700 800 900 1000 300 400 500 600 700 800 900 1000
Wavelength μm Waveband

Fig. 1. Effect of inoculum levels of Meloidogyne incognita, on Fig. 2. Effect of management options on the reflective pattern
the reflective pattern of light from plant leaf canopy in of light from plant leaf canopy in mungbean cv. Pusa Vishal
mungbean cv. Pusa Vishal after 25 days of inoculation

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Indian Journal of Nematology Vol. 43, No. 1

the use of remote sensing of sybean cyst nematode with Gausman, H.W., Heald, C.M. & Escobar, D.E. (1975). Journal
infra red reflectance through landsat and observed 60% of Nematology 7: 368-374.
variation in data.
Lawrence, G.W., Doshi, R.A., King, R.L., Lawrence, K.S. &
Caceres, J. (2007). Nematode management using remote
Results indicate the potential for remotely sensing to sensing technology, self-organized maps and variable
detect nematode-infested plants. rate nematicide applications. World Cotton Research
Conference-4, Lubbock, Texas, USA, 10-14 September
REFERNCES 2007 pp. unpaginated

Chrishtian, H., Astrid, S., Volker, K. & Richard A.S. (2010) Nutter, F.W. Jr., Tylka, G.L., Guab, J., Moreira, A.J.D.,
Remote sensing for detection of soil born plant parasitic Marett, C.C., Roseberg, T.R.B., Asart, J.P. & Chong,
nematode and fungi pathogen. Precision of crop C.S. (2002). Journal of Nematology 34: 222-231.
protection –the challenge and use of heterogeneity. E C
Oerke et al edition chapter 10 Springer science and Palacharla, P.K., Durbha, S.S., King, R.L., Gokaaraju, B. &
business Media. Lawrence, G.W. (2011). IEEE 978: 249-252.

Diversity of Plant Parasitic Nematodes in Banana at Ten Districts of Assam

DEBANAND DAS*, ALPONA DEURI AND KARTIK BARUAH1

Department of Nematology, 1Department of Horticulture, Assam Agricultural University, Jorhat-785 013

*Corresponding author, E-mail: debanand2009@[Link]

Received on 10-04-2013 and Accepted on 30-04-2013

Banana and plantains constitutes the fourth most Keeping the view of limiting production and
important fruit crops in the tropics and subtropics forming productivity of banana by nematodes the present
major constituents of diet for millions of people across investigation was carried out to examine the biodiversity
the globe. of plant parasitic nematodes around banana rhizospheres
in ten districts of Assam.
Production of banana is challenged by many pests
and diseases, of which plant parasitic nematodes are Roving survey was conducted at ten banana growing
considered as countervail. Globally, a total of 151 species districts of Assam during 2006-2011. Plant as well as
of nematodes belonging to 51 genera have been root samples were collected from around the rhizosphere
documented to be associated with banana rhizosphere of banana at around 30 cm away from the bole of the
(Gowen & Queneherve, 1990; Koshy & Gulsar Banu, plant and at a depth of 15-50 cm. From each site a total
2000). In India, 71 species of plant parasitic nematodes of 5 subsamples were collected, and then these
are known to be associated with banana (Krishnappa & subsamples were mixed thoroughly to draw a
Reddy, 1995; Koshy & Sosamma, 2001). Yield loss homogeneous sample of 500 gram. 10 gram of root
caused by nematodes to bananas are very high, with sample also collected from each site. Thus, a total of 150
average annual yield losses estimated at about 20 per samples were collected from nine districts of Assam.
cent worldwide (Sasser & Frackman, 1987), but may Soil samples (200cc of soil from each sample) were
reach up to 40 per cent or more when frequent tropical processed by following Cobb’s modified sieving and
storms topple plants with rotten, nematode infested root decanting technique and root samples (5 gm of root from
system. each sample) were processed by Baermann Funnel

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Table 1: Plant parasitic nematodes associated with banana rhizosphere at different districts of Assam.

Districts Population of different nematodes (200cc of soil)

Helicotylenchus Meloidogyne Pratylenchus Rotylenchulus Tylenchorhynchus Macroposthonia sp

dihystera incognita thornei reniformis leviterminalis

Nalbari 88 – 116 48 – 85 18 – 46 24 – 50 19 – 64 12
(80.25) (56.25) (37.50) (43.75) (50.0) (13.33)
Borpeta 52 – 172 52 – 98 25 – 54 28 – 56 16 – 54 7
(75.0) (60.0) (50.0) (37.5) (31.25) (6.67)
Cachar 35 -160 26 – 52 10 -28 10 – 62 23 – 82 5
(80.0) (55.0) (30.0) (50.0) (30.0) (11.33)
Karimganj 25 – 170 12 – 40 4 – 16 9 – 18 9 – 46 7
(70.0) (45.0) (35.0) (50.0) (35.0) (10.0)
Hailakandi 35 – 138 12 – 52 4 – 18 6 – 22 8 – 60 5-12
(75.0) (50.0) (40.0) (45.0) (35.0) (10.0)
Karbi Anglong 56 – 161 43 – 76 34 – 85 77 – 121 32 – 121 13-20
(85.0) (60.0) (35.0) (40.0) (40.0) (30.0)
Lakhimpur 85 – 162 25 – 52 38 – 88 — 24 – 50 18-27
(100.0) (56.0) (20.0) (43.0) (25.0)
Dhubri 52 – 142 46 – 54 32 – 98 25 – 54 28 – 56 7-16
(100.0) (70.0) (15.0) (21.0) (41.0) (19.0)
Golaghat 121 – 156 32 – 65 — 33 – 43 21 – 33 12-45
(100.0) (60.0) (26.33) (60.0) (80.0)
Jorhat 32 – 220 12 – 76 10 – 78 25 – 88 7 – 91 23-35
(100.0) (66.67) (35.67) (28.55) (62.0) (26.33)

(Data in parenthesis are the per cent frequency of occurrence)

technique (Christei and Perry, 1951). Nematodes were reniformis, Tylenchorhynchus leviterminalis,
identified upto generic level under stereozoom binocular Hoplolaimus sp., Macroposthonia sp., Psilenchus
microscope and counting of nematodes were made by sp., and Longidorus sp. Psilenchus sp and Lonidorus
using a Hawkshly Nematode counting dish. Identification sp. has been reported for the first time from around the
of nematodes up to species level was done under rhizosphere of banana in Assam. Study revealed that
stereozoom research microscope. Frequency of Helicotylenchus dihystera was the most predominant
occurrence was calculated following Norton’s (1978 ) nematode species in all the districts surveyed, with 80 –
formula. 100 per cent frequency of occurrence. The gall forming
nematode, Meloidogyne incognita was found to occur
All the 250 soils and root samples were found in 55-70 per cent of the samples. Another important
positive for plant parasitic nematodes. A total of eight nematode, Pratylenchus thornei occurred in 15 – 50
different plant parasitic nematodes (Table 1), belonging per cent of the samples.
to eight genera were recorded from the rhizosphere of
banana plantations of ten districts of Assam. Plant Population of H. Dihystera was found more in all
parasitic species recorded during the present investigation the districts, irrespective of the banana cultivar. Singh &
are: Helicotylenchus dihystera, Meloidogyne Uma (1996) also recoded high population of this nematode
incognita, Pratylenchus thornei, Rotylenchulus irrespective of growing season and variety. Root-knot

103
Indian Journal of Nematology Vol. 43, No. 1

nematode, M. Incignita was found to produce medium Gowen, S.R. & Queneherve, P. (1990). Nematode parasites of
root gall on the root system of banana. During the visual bananas, plantains and abaca. In ‘Plant parasitic
nematodes in subtropical and tropical a g r i c u l t u r e ’
observation, no external plant symptoms were noticed on
(eds. M. Luc, R.A. Sikora, J. Bridge). CABI Publication
plants affected by root knot nematode. The attack of M. Wallingford, UK, 648pp.
Incognita alone can cause a loss of 30.95 per cent to
yield of banana (Jonathan & Rajendran, 2000),while in Jonathan, P.I. & Rajendran, G. (2000). Indian Journal of
association with Fusarium the loss may be higher. Nematology 30: 162-164.

Population of lesion nematode, Pratylenchus thornei Khan, M.R. & Hasan, M.A. (2010). Journal of Plant Protection
was recorded at lower level and this species was not Research 50: 263-268.
recorded from Golaghat district during the present survey.
Koshy, P.K. & Gulsar Banu, J. (2000). Nematode diversity in
Characteristic dark lesion on the cortical region of root plantation crops: the world scenario. In “Nematode
was observed, mostly on cv. ‘Jahaji’. The present diversity” (ed. M.S. Jairajpuri). Maulana Azad National
investigation revealed the multi nematode species Urdu University, Hyderabad, India, 500p.
infestation in the banana plantation. Khan and Hasan
(2010) also observed similar kind of nematode infestation Koshy, P.K. & Sosamma, V.K. (2001). Nematode diseases of
on banana in West Bengal. They recorded eight different plantation crops andtheir management. In ‘National
Congress on Centenary of Nematology in India-appraisal
genera of plant parasitic nematodes from the banana
future plans’. Division of Nematology, IARI, New Delhi,
rhizosphere. 5-7 December, 2001. 176 pp.

The investigation on nematode diversity around Krishnappa, K. & Reddy, B.M.R. (1995). Nematode problems
banana rhizosphere indicated that banana roots are of banana in India. In ‘ Nematode pest management –
prone to nematode attack and they co-inhibit a large an appraisal of ecofriendly approaches’ (eds. G. Swarup,
density forming a nematode species complex. The D.R. Dasgupta, J.S. Gill) Nematological Society of India,
New Delhi, India, 300pp.
information generated through this investigation will be
helpful to provide advisory services and to undertake Norton, D.C. (1978). Ecology of plant parasitic nematodes.
pest risk analysis. Wiley & Sons, New York, 147 pp.

ACKNOWLEDGEMENT Sasser, J.N. & Frackman, D.W. (1987). A worlds perspective

on Nematology: the role of Society. In ‘Vistas on
The work was carried out under the financial support Nematology’ (eds J.A. Veech, D.W. Dickson) Hyattville,
of AICRP on Tropical Fruits, IIHR, Bangalore. MD, Society of Nematologist. 509 pp.

REFERNCES Singh, H.P. and Uma, S. (1996). Banana cultivation in India.

Directorate of Extension, Ministry of Agriculture,
Government of India, Krishi Bhawan, New Delhi, India,
Christei, J.R. & Perry, V.G. (1951). Removing nematodes
102p.
from soil. Proceedings of Helminthology Society
Washington 18: 106-108.

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 Short Communication

Screening of Cucumber Cultivars Against Meloidigyne incognita

NIKHITA KAKATI AND B. MAHANTA

Department of Nematology, Assam Agricultural University, Jorhat-785 013, India

*Corresponding author, E-mail:

Received on 16-03-2013 and Accepted on 20-04-2013

Root-knot nematode, Meloidogyne incognita is a Table 1. Screening of cucumber cultivars against

polyphagus pest of vegetable crops including cucumber Meloidogyne incognita
(Cucumis sativus L.) causing significant reduction in Mean of 5 replications
plant growth and fruit yield. Krishnaveni and Subramanian
(2002) recorded 69.2 per cent yield loss in cucumber due Cultivars Mean root-knot Reaction
index (1-5 scale)
to M. incognita. Therefore, effort was made to screen
varieties of cucumber for their resistance against this EC 641913 2.4 MR
major nematode pest. Darekar and Bele (1990) observed
EC 641908 3.6 S
that with an increase in the inoculums level of M.
incognita, there was a progressive decrease in growth EC 641912 4 S
parameters of cucumber seedlings. In the present EC 641920 3.8 S
investigation, 18 varieties of cucumber were screened.
EC 641927 4 S
In the present study 7 varieties were supplied from the
NBPGR and others were collected from local market. EC 641934 3.6 S
Cucumber green long special 3.4 S
Three seeds of each varieties were sown in 10 cm
diameter earthen pots containing 500 g starilized soil. Malini 4 S
After one week of germination one healthy seedling per Bankim 3.4 S
pot was retained. Three week after sowing, 500 freshly
Cucumber No. 243 3.6 S
hatched second stage juveniles of M. incognita were
inoculated. Each treatment was replicated five times and Nandini 4 S
pots were arranged in Completely randomized design. Cucumber No-786 3.6 S
After 45 days of inoculation, the plants were uprooted
Cucumber green long (Check) 4 S
and root system was washed gently. Number of galls and
egg masses per root system were counted. The gall index EC 641925 4.4 HS
was recorded as per the 1-5 scale as highly resistant Kalyan 4.6 HS
(0.00), resistant (0.01-2.00), moderately resistant (2.01-
3.00), susceptible (3.01-4.00) and highly susceptible Cucumber NS-408 4.6 HS
(4.01-5.00). Debstar 4.4 HS
Alisa 4.6 HS
Out of 18 cultivars, none of the cultivars ranked in
resistant reaction. Only one as moderately resistant (EC Improve Noori 4.2 HS
641913), eleven were susceptible (EC 641908, EC 641912, [Link] 0.29
EC 641920, EC 641927, EC 641934, Cucumber green
CD (P=0.05) 0.58
long special, Malini, Bankim, Cucumber No. 243, Nandini,
Cucumber No-786) and six cultivars were found to be MR : Moderately Resistant; S: Susceptible; HS : Highly
highly susceptible (EC 641925, Kalyan, Cucumber NS- Susceptible
408, Debstar, Alisa, Improve Noori).
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Indian Journal of Nematology Vol. 43, No. 1

REFERENCES Krishnaveni, M. & Subramanian, S. (2002). Assessment of

yield loss in cucumber (Cucumis sativus L.) due to
Darekar & Bele (1990). International Nematology Network Meloidogyne incognita. Proceedings of National
Newsletter 7: 17-18. Symposium on Biodiversity and Management of
Nematodes in Cropping Systems for Sustainable
Agriculture, Jaipur, India, 11-13 November, pp. 88-89.

Effect of Different Plant Extracts as Root Dip Treatment Against Root-Knot

Nematode, Meloidogyne incognita Infecting Tomato
MANJU MEENA, S. BHARGAVA, M.K. SHARMA AND H.R. GURJAR*

Department of Nematology, Rajasthan College of Agriculture, Udaipur-313 001, India

*Corresponding author, E-mail: hemrajg7@[Link]

Received on 21-05-2013 and Accepted on 21-05-2013

Tomato [Lycopersicum esculentum Mill] is one of extract at 70° C till it become a semi-solid material. This
the most important annual vegetable crop grown semi-solid material becomes stock solution by adding
throughout the country. Among various pest and diseases, distilled water and makes different respective
which damage crops, plant parasitic nematodes, present concentration by different plant extracts stock solution.
a formidable pest problem for different crops. Most
species attack and feed on plant roots and underground In-vivo effect of plant extracts on the root-knot
plant parts. Vegetable crops are excellent host for a wide nematode infecting tomato: Tomato seedling were raised
range of plant parasitic nematodes. Estimated overall in bulk and seedling of uniform size were dipped in
average annual yield losses of the world major vegetable alcoholic all there plant extracts along with best two
crops by nematodes are 12.3 per cent. Average losses concentration of each with one untreated check and one
for the 40 crops in developed countries were estimated chemical check i.e., Carbosulfan 25 EC 2 per cent for 4
to be 8.8 per cent compared with 14.6 per cent for hrs. The treated seedlings were transplanted in pot. The
developing countries (Ravichandra, 2008). experiment was laid out in CRD with four replications.

So far substantial work has been done on various Three tomato seedlings were sown in each pot.
aspects of root-knot nematode, M. incognita on tomato After 10 days, one healthy plant in each pot was maintained
but there is not much information available on management and others were uprooted carefully. The pots were
of root-knot nematode through plant extracts. watered regularly as and when required. Observation on
plant growth and reproduction of root- knot nematode on
Healthy leaves of neem (A. indica), karanj (P. tomato were recorded at 45 days after transplanting or
pinnata), castor (R. communis) and leaves of jatropha at harvest.
(J. curcas) were collected from herbal park of RCA,
Udaipur (Raj.). The collected plant parts were washed Observation on shoot length (cm), root length (cm),
with distilled water. Hundred gram of clean fresh plant fresh shoot weight (g), and fresh root weight (g) were
leaves were grind with 100 ml absolute alcohol. The taken at harvest. The root were washed carefully under
mixture was allowed to stand for 48 hrs at room tap water and stained with 0.1 per cent acid fuchsin lacto
temperature and was subsequently filtered through filter phenol (Mc Beth, 1941) and after wash kept in clear
paper. The solvent was completely evaporated from the lacto phenol for 24 h. Thereafter the roots were examined

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 Short Communication

thoroughly under a stereoscopic binocular microscope eggmass-l (190.75), larval population 100 CC-l soil (233)
for counting number of gall per plant, number of egg and total population (6469) was recorded in plants treated
masses per plant and number of eggs and larvae per egg with 20% Neem plant leaf extract (T4), followed by 20%
mass. After removing the plant from the pot, soil was Jatropha plant leaf extract (T2), 20% Karanj plant leaf
thoroughly mixed and 100cc soil from each pot were extract (T6) and 20% Castor plant leaf extract (T8) over
taken and processed by Cobb’s sieving and decanting untreated check (Table 1). The results of the study
technique (Cobb, 1918) followed by Baermann’s funnel clearly indicated that the treatments effects were better
technique (Christie and Perry, 1951) for estimation of realized with 20% Neem plant leaf extract.
nematode population in soil. The data were subjected to
statistical analysis. To find out the eco-friendly and economically feasible
method for management of root- knot nematode,
Tl= 10% Jatropha plant leaf extract; T2= 20% Meloidogyne incognita infecting tomato. All the four
Jatropha plant leaf extract; T3= 10% Neem plant leaf plant leaf extracts namely neem (Azadirachta indica),
extract; T4= 20% Neem plant leaf extract; T5= 10% karanj (Pongamia pinnata), castor (Ricinus communis)
Karanj plant leaf extract; T6= 20% Karanj plant leaf and jatropha (Jatropha curcas) with two concentrations
extract; T7= 10% Castor plant leaf extract; T8= 20% (10 & 20%) used as root dip treatment found superior
Castor plant leaf extract; T9= Chemical check over untreated check (control) in improving plant growth
(Carbosulfan) 2%; T10= Untreated check. An and reducing nematode reproduction. However, leaf
experiment with 10 treatments was conducted in CRD extract ofneem (Azadirachta indica) at 20 per cent
with four rel?lications. was found to be the best treatment in improving plant
growth characters and in reducing nematode reproduction
The observation on plant growth characters and on tomato followed by leaf extracts of jatropha (Jatropha
reproduction of M. incognita on tomato are presented curcas), karanj (Pongamia pinnata) and castor (Ricinus
in (Table 1). communis) at 20 per cent concentration.

Results of the experiments conducted during kharif, This investigation is in adjustable conformity with the
2012 at Rajasthan College of Agriculture, Udaipur to see finding of Saravanapriya & Sivakumar (2003) that tested
the effect plant leaf extracts of neem (Azadirachta the efficacy of leaf extract of some botanicals namely
indica), karanj (Pongamia pinnata), castor (Ricinus Calotropis gigantea, Tagetus erecta, Azadirachta
communis) and jatropha (Jatropha curcas) are presented indica, Areca catechu and Citrullus lantatus against
Data pertaining to the effect of plant leaf extracts on the root-knot nematode, M. incognita infecting tomato.
plant growth characters (shoot length and weight, root The leaf extract of C. gigantea significantly reduced the
length and weight) and nematode reproduction (number nematode population both at 45 days af!er transplanting
of galls per plant, number of egg masses per plant, (87.30%) and at harvest (89.96%) over control. The
number of eggs and larvae per egg mass, soil population same treatment also increased fruit yield by 23.91 per
per 100cc soil and total population of nematodes) were cent. Reported by Sosamma & Jayasree, (2002).
recorded and statistically analyzed to test the significance Different plants including jatropha have different type of
of results. saponin and exhibits different type of biological activity
.These saponins are being successively used as larvicidal
All the treatments were effective in significantly and nematicidal because of its ampililic nature. Omar et
reducing the nematode population and increasing plant al. (1994) reported that Q. saponaria saponin wextract
growth. Among the individual treatments, significant at 260-280 ppm reduced the number of egg masses and
increase in shoot length (31.61 cm), shoot weight (20.67 viable juveniles of the root-knot nematode, M. javanica.
g), root length (24.13 cm), root weight (3.38 g) and Hence, in the present study leaf extract of neem
significant decreases in number of galls plant-l (18.46), (Azadirachta indica), jatropha (Jatropha curcas),
no. of egg masses plant-l (21.70), no. ofeggs & larvae karanj (Pongamia pinnata) and castor (Ricinus

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 Table 1. Effect of Plant Leaf Extracts as Root Dip Treatment against Root-knot Nematode, Meloidogyne incognita Infecting Tomato Under Pot Experiment.

Treatments Plant Growth Characters Nematode Reproduction

Shoot Shoot Root Root No. of No. of No. of larval Total
Length Weight Length Weight galls/ egg eggs & population population
(cm) (g) (cm) (g) plant masses/ larvae/ in 100 cc of
Indian Journal of Nematology Vol. 43, No. 1

plant egg mass soil nematode

10% Jatropha plant leaf extract T1 24.38 13.66 18.45 1.86 32.83 36.25 217.50 262 10510
20% Jatropha plant leaf extract T2 29.12 18.25 21.82 2.23 21.83 24.50 191.75 244 7141
10% Neem plant leaf extract T3 25.79 15.38 19.88 1.94 30.92 35.50 213.00 257 10134
20% Neem plant leaf extract T4 31.61 20.67 24.13 3.38 18.46 21.70 190.75 233 6469

108
10% Karanj plant leaf extract T5 22.96 13.22 16.17 1.61 35.92 36.25 228.75 268 10971
20% Karanj plant leaf extract T6 29.80 17.25 20.85 2.15 23.10 25.25 207.75 249 7729
10% Castor plant leaf extract T7 24.23 14.08 18.58 1.83 33.33 35.25 220.00 262 10363
20% Castor plant leaf extract T8 29.16 18.21 21.92 2.17 21.33 27.25 201.00 245 7923
Chemical check (Carbosulfan) 2% T9 35.25 23.30 31.89 3.44 16.25 8.50 134.50 222 3363
Nematode alone (Control) T10 22.25 9.25 14.18 1.39 48.23 46.00 207.75 300 12558
[Link]± 0.954 0.467 0.513 0.099 0.513 0.798 4.426 6.997 221.623
CD (P=0.05) 2.756 1.348 1.481 0.285 1.481 2.304 12.783 20.209 640.094

Note: (i) Data are average value of four replication (ii) Initial inoculums 2 larvae/g soil.
 Short Communication

communis) at higher concentrations were found most Ravichandra, N.G. (2008). Plant Nematology I.K. International
effective in reducing population of M. incognita on Pulishing house, New Delhi. 110016.
tomato and in enhancing the plant growth characters.
Sarvanpriya, B. & Sivakumar, M. (2003). Effect of plant
extracts on tomato against Meloidogyne incognita.
REFERENCES Proceedings of National Symposium on Biodiversity
and Management of Nematodes in Cropping Systems
Christie, J.R. & Perry, V.G. (1951). Proceedings of for Sustainable Agriculture, Jaipur, India, 11-13
Heminthological Society of Washington 18: 106-108. November, 2002-2003: 125-128.

Mc Beth, C.W., Taylor, A.L. & Smith, A.L. (1941). Proceeding Sosamma, V.K. & Jayasree, D. (2002). Indian Journal of
of Helminthological Society of Washington 8: 26. Nematology 32: 183-233.

Omar, S.A., Abdel-Massih, M.I. & Mohamed, B.E. (1994).

Bulletin of Faculty of Agriculture of Cairo 45: 933-940.

Community Analysis of Plant Parasitic Nematodes Associated with Rice in

Imphal East District of Manipur (Pourabi)
L. JOYMATI*, KH. RONIBALA, ZENITH NG. AND P. PURNIMA

Nematology Laboratory, Department of Zoology, P.G. Centre, D.M. College of Science, Imphal-795 001, India
*Corresponding author, E-mail: drjoymatidevi@[Link]

Received on 26-11-2012, Resubmitted on 04-01-2013 and Accepted on 26-01-2013

Rice is one of the valuable cereal food crop in the occurrence of plant parasitic nematodes associated with
world and forms the staple food for more than half of the rice.
global population. In India, rice is grown in almost all the
state as a principal food crop and occupies about 23.3% The present survey was carried out in eighteen
of the gross cropped area of the country , covering about different villages of Pourabi, Imphal East from August
44.8 mha of land. In Manipur, it occupies about the 2012-0ctober 2012 during summer season namely: Laijing
maximum cultivated area (Anon, 2008). Rice contributes maya loubuk, Laijing ching maya loubuk, Sinam lok
43% of the total food grain production and 46% of the loukon, Yomalok loukon, Ehickon loukon, Tairenpokpi
total cereal production in India. Several soil and plant loukon, Kharou loukon, Kanglou loukon, Ikop loukon,
inhabiting parasitic nematodes have been known to be Thingel makha loukon, Kongon toubi loukon, Nungoi
associated with vegetables, crops (Roy, 2005). Plant loukon, Huidrom loukon, Sawombung loukon, Hidumbi
parasitic and free living nematodes are the important loukon, Bamon loukon, Patlou loukon, Wairi loukon,
members of the nematodes community and it cause Maniyaiskul loukon and Thamnapokpi loukon. Diseased
serious damage to many fruits, vegetables, crops etc. field were selected on the base of above ground symptoms
The annual yield loss in rice due to plant parasitic of the crops such as, wilting, slow growth, stunting and
nematodes on a world basis is estimated to be 10% yellowing of leaves. Soil sample from the associated
(Sasser & Freckrnan,1987). The present survey of rice rhizosphere were collected from a depth of 10-15 cm at
growing area of Imphal East district of Manipur was the rate of one unit sample. Each unit sample was a
taken from August 2012- October 2012 for recording the composite of 20 cores obtained from four comers and

109
 Table 1: Occurrence of plant parasitic nematodes within Pourabi in Imphal east district of Manipur

Village Crop Nematode population/200 cm3 soil

Meloidogyne Helicotylenchus Tylenchorhynchus Pratylenchus Hoplplaimus Xiphinema Criconematode

graminicola multieinetus sp. sp. sp. sp. sp.

Loijing maya Thangjing 50 - 87 43 - - -

loubuk
Loijing ching Kd angoubi - 946 - - - - -
Maya loukon
Sinam lok Kd angoubi 44 230 - 54 - 34 -
loukon
Yomalok loukon Phaungang - - 115 - 178 - 290
Indian Journal of Nematology Vol. 43, No. 1

Checkon loukon Chabokpi 12423 - - 69 100 - 250

Kanglou loukon Ayangleima 6614 500 - 45 - 65 -
Ikop loukon Hemochandra - - 700 - 850 - 24
Thingel makha Dharam 48 - 100 - - - -
loukon
Tairenpokpi Jatra phou - 4400 - 388 - 450 -

110
loukon
Kharou loukon Nongin 135 - 550 - 450 - 800
Kongon toubi Khamba - - - 750 - 500 -
loukon
Nungoi loukon Phoudum 200 100 - - 250 - 300
Huidrom loukon Jatra phou - 2065 - - 308 - -
Sawombung Dharam 85 - 1020 - - - -
loukon
Hidumbi loukon KD angoubi - 125 - 560 - 625 -
Wairikhul loukon Phaungang - - - 1012 - - 560
Mani yaiskul Chabokpi 3400 - - - 540 - 100
loukon
Thamnapok pi Hemochandra 25 - 150 - - 5 -
loukon
Bamon loukon Thangjing 120 - - 50 - - 400
Patlou loukon K Damubi - 1150 - - 50 - 100
 Short Communication

centre of the field. Root soil sub sample were stored in phou at Tairenpokpi loukon and lowest density (50 J2s/
polythene bags and kept at 4°C in a refrigerator not more 500g of soil) in Hemochandra and Khamba at Ikpo
than 7 days. Altogether 180 soil samples and roots were loukon and Kongon loukon. Tylenchorhynchus spp.
collected, 10 samples from each field. Infected roots showed highest density of 1020 J2s/500g of soil in
were stained in cotton blue-lactophenol and observed for Dharam at Sawombung loukon and lowest of 87 J2s/
the presence of nematodes. The females of root-knot 500g of soil in Thangjing variety of rice at Laijing maya
nematodes were dissected out from the galled roots and loukon of Pourabi. Among these plant parasitic nematodes,
perineal section prepared for species identification. The Hoplolaimus spp., Xiphinema spp. and Criconematodes
estimation of nematode population per 10 9 root samples spp. were found to be in more or less same number of
and 2009 soil sample was done in a multi-chambered population density in different variety of rice at different
counting dish under a stereoscopic binocular microscope. localities of [Link] for Pratylenchus spp. highest
The population densities of nematode species in the density (1012 J2s/500g of siol) was found in Phaungang
samples were calculated using the formulae (Norton, at Laijing maya loukon of Pourabi.
1978).
Table 2 shows, analysis of soil and root samples of
Seven plant parasitic nematode genera viz., different rice variety of Pourabi revealed that most of the
Meloidogyne graminicola, Helicotylenchus collected smaples were severely infested with seven
multicinetus, Tylenchorhynchus spp., Pratylenchus nematodes viz., Meloidogyne graminicola,
spp., Hoplolaimus spp., Xiphinema spp. and Helicotylenchus multicinktus, Tylenchorhynchus spp.,
Criconematodes spp. were found associated with Pratylenchus spp., Hoplolaimus spp., Xiphinema spp.
different varieties of rice in different localities of Pourabi. and Criconematodes spp. Among these root-knot
Highest number of Meloidogyne graminicola (12,423 nematodes Meloidogyne graminicola occurred most
J2s/500g of soil) was recorded in Chabokpi variety of frequently followed by Helicotylenchus multicinktus
rice at Checkon loukon of Pourabi with lowest density with absolute frequency of 6.11 % and 5.55% respectively.
(44 J2s/500g of soil) was recorded in KD am ubi at Sinam The root-knot nematode, M. graminicola was found in
lok loukon of Pourabi (Table 1). The population density higher density (18.96%) followed by Helicotylenchus
of Helicotylenchus multicintus was found to be slightly multicinktus (17.24%). M. graminicola is the most
lower as compared to M. graminicola. Its highest predominant phytonematodes of rice.
density (4400 J2s/500g of soil) was recorded in Jatra
The results support the report of Joymati & Thoithoi
Table 2: Community analysisi of plant parasitic nematodes (20 10) about the incidence of rice root-knot nematode
in different villages of Pourabi, Imphal east district M. graminicola at Thoubal district of Manipur. The
of Manipur work also support the of Joymati et al., (2011 ) about the
Nematode Absolute Absolute Relative
community analysis of plant parasitic nematode in
frequency density frequency different vegetables field of Bishnupur district of Manipur.
The present findings can give important knowledge
Meloidogyne 6.11 661.25 18.96 about plant parasitic nematodes especially rice root-knot
graminicola
nematode which cause serious damage to food crop i.e.,
Helicotylenchus 5.55 274.74 17.24 rice and leads to totally damaged in the production of rice
multicinktus in the state.
Tylenchorhynchus spp. 3.88 77.77 12.06
ACKNOWLEDGEMENT
Pratylenchus spp. 5 90.6 15.51
Hoplolaimus spp. 4.44 77.88 13.79 The author greatly acknowledged D.S.T., New
Xiphinema spp. 3.33 62.11 10.34 Delhi for financial assistance and Principal, D.M. College
of Sciences for providing Laboratory facilities during the
Criconematode spp. 5 80.68 15.51
course of studies.

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Indian Journal of Nematology Vol. 43, No. 1

REFERENCES Roy, K. (2005). Studies in the role of reniform nematode,

Rotylenchus reniformis Linford & Oliveira in vegetable
Anonymous (2008). Directorate of Economics & Statistic Govt. crop production in West Bengal. Ph.D. Thesis
of Manipur. Statistical Abstract Manipur 2008 Department of Agricultural Entomology, B.C.K.V.,
Mohanpur, Nadia. Wset Bengal pp. 38-87.
Joymati, L., Christina, Kh., Jennifer, O. & Bibi, K. (2011).
Indian Journal of Nematology 41: 220-221. Sasser, J.N. & Freckman, D.W. (1987). In: Vistas on
Nematology (Eds., J.A. Veech & Dickson, D.W.) p 7-14.
Joymati, L. & Thoithoi, M. (2010). Indian Journal of Hyattsuille, USA. 509 pp.
Nematology 40: 231.

Norton, D.C. (1978). Ecology of plant parasitic nematodes.

John Wiley, New York. pp.266.

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 Book Review

BOOK REVIEW
Nematodes of Banana: Taxonomy and Ecology of Dorylaimids and Tylenchids by Viswa V.
Gantait, T .Bhattacharya and [Link] Published by Lambert Academic Publishing Germany, (2010,
pp. 300)
The book contains the details of the work done carried out on the survey of soil and plant parasitic nematodes belonging
to the order Dorylaimida and Tylenchida associated with Banana plantations(Musa paradisica [Link] Kanthali) in
Pashim Medinipur District of West Bengal, [Link] second aspect deals with the study of community structure of
durylaimids and tylenchid associated with banana [Link] third aspect concerns with the temperal variation of
four selected edaphic factors. The last one revolves around the population dynamics of nematode population in relation
to the edaphic factors.
During the survey a total 46 species of soil and plant parasitic nematodes belonging to 32 genera and 15 families under
order Dorylaimida and Tylenchida have been collected and identified amongst dorylaimids, 8 species and amongst
tylenchids 4 species are recorded as new. The species key of the five genera viz. Dorylaimus Lainydorus,
Lagenonema, Makatinus and Discolaimus are provided. Sixteen species (12 dorylaimus and 4 tylenchids) are recorded
for the first time from the rhzosphere of banana. Community composition of nematodes was analysed with reference
to abundance, frequence of occurrence, relative abundance, mean crowding, dominance index species diversity and
species evenness.
Among the tylenchids, Rotylenchulus reniformis was the most abundant and Pratylenchus coffeae was least
abundant species. Among dorylaimids, Prodorylaimus sukuliwas the most abundant and Promuntazium elongatum
was the least abundant species. This book therefore, provides the valuable information about the taxonomy of
dorylaimid and tylenchid nematodes of banana in India ,their community structure population dynamics and seasonal
population fluctuation in relation to different edaphic factor. Hence, it will serve as reference book for the researchers
working in banana.
Nematodes Associated with Guava and Litchi Plantations: Studies on Taxonomy and Ecology of Soil and
Phytophagous Nematode by Oebabrata Sen,Amalendu Chatterjee and Buddhadeb Manna; Lambert
Academic Publishing Germany (2012, pp. 321).
This book is outcome of comprehensive study on soil and plant parasitic nematodes belonging to the orders Dorylaimida
and Tylenchida associated with Guava and Litchi plantations in India. Guava and Litchi are the most economically
important fruit crops in West [Link] book dealt with taxonomy and ecology of soil and plant parasitic nematodes
belonging to the orders Dorylaimida and Tylenchida associated with these crops. Thirty one species of Dorylaimida
under 19 genera,14 subfamilies,10 families, 5 superfamilies and 2 suborders have been observed. From the study, one
genus and 11 species were added to the science. In addition, two species from India, nine species from West Bengal
and eight species from south 24 Parganas has been recorded for the first time. Under the order Tylenchida one genus
and 3 species have been proposed as new. Different edaphic factors, like soil moisture, temperature, pH, nitrogen,
phosphorus, potassium, organic carbon and soil electrical conductivity were estimated as they are determining factors
for the shape and quality of the nematode population. The population fluctuation of phytophagous nematode were
observed in relation to predatory and omnivorous nematodes belonging to the orders Mononchida and Dorylaimida.
This book therefore provides the valuable information about the taxonomy of dorylaimid and tylenchid nematodes of
Litchi and Guava plantations in India, Hence it will serve as reference book for the researchers working on Guava &
Litchi.
Pankaj and [Link]
Division of Nematology
Indian Agricultural Research Institute
New Delhi-110012

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Indian Journal of Nematology Vol. 43 No. 1

NEMATOLOGICAL NEWS
● Dr. H.S. Gaur, former JD (Edn.) & Dean, PG School, IARI, New Delhi has joined as Vice Chancellor, Sardar
Vallabh Bhai Patel University, Meerut on 16th Feb., 2013.

● Dr. Viswa Venkat Gantait from Zoological Survey of India, Kolkata participated in the 32nd ISEA as a summer
member during November 2012 to April 2013. He was the only Biologist in the expedition group. He was nominated
as a Zoologist, especially as a Nematologist, as because the nematodes occupy about 22% of the total invertebrate
fauna of Antartica. In the field of Nematology, he is the first in India who attended the Indian Scientific Expedition
to Antartica. Dr. V.V. Gantait stayed in both the stations (Dakshin Gangotri and Maitri) and collected more than
100 soil, moss and algal samples from different locations of 'Larshman Hills', 'Schirmacher Oasis'. All the samples
have been collected for taxonomic and ecological studies of invertebrate fauna in Antartica.

● Department of Nematology, CCS Haryana Agricultural University and KVK, Fatehabad of this university
organised a Molya Gyan Diwas at village Khabra kalan in district Fatehabad on 20th October,2012. Dr Ravi
Kumar, AD (ResearchExtension Linkage) of University was the chief guest on this occasion. About 150 farmers
of Khabra kalan and surrounding villages and ofificers of agriculture department of Fatehabad district participated
in this Gyan Diwas. On 20th March, 2013, a Molya Khet diwas (field day was organised in village Salam Khera
(Fatehabad). Dr [Link], Associate Director (Farm Advisory Services), CCS HAU Hisar graced this
occasion. It was attended by ca. one hundered farmers, agriculture and horticulture officers of the district. Purpose
of these programmes was to awaken the farmers and field functionaries about the crop diseases caused by
nematodes particularly Molya disease in wheat and barley. Dr K.R. Dabur, Professor and Head , DR.I.J. Paruthi,
Dr. [Link] (Department of Nematology) and Dr. [Link] (KVK, Fatehabad) delievered expert lectures
on nematode problems of cotton, wheat, guar and horticultural crops.

● Dr. (Mrs.) Padma Bohra, a well known taxonomist in Nematology, Desert Regional Centre, Zoological Survey
of India, Jodhpur passed away on 27th May, 2013. She was awarded Ph. D. degree from JNV University, Jodhpur
on the thesis entitled "Taxonomy of Plant and Soil Nematodes Associated with Cereal Crops in Rajasthan" under
the able guidance of Dr. Qaiser H. Baqri in the year 1999. She was selected to the post of Scientist-C in the year
2002 and was subsequently promoted to the post of Scientist-D in the year 2008. She also served as Officer-in-
Charge of Desert Regional Centre, ZSI, Jodhpur from August, 2006 till her last breath. During her short carrer
in research on taxonomy of plant and soil nematodes (1996-2013), she described one new genus and more than
12 new species, besides recording more than 100 species for the first time from Rajasthan and Gujarat states and
several species for the first time from India in the journals of national as well international repute.

114