Journal of Chemical Ecology, VoL 20, No.

12, 1994

INHIBITION OF SALVINIA (Salvinia molesta MITCHELL)

BY PARTHENIUM (Parthenium hysterophorus L.). II.
RELATIVE EFFECT OF FLOWER, LEAF, STEM, AND
ROOT RESIDUE ON SALVINIA AND PADDY

D.K. PANDEY

National Research Centre fi~r Weed Science (I. C.A.R. ),

Adhartal, Jabalpur (M.P.)-482 004, hldia

(Received February 23, 1994; accepted July 25. 1994)

Abstract--The relative effect of parthenium (Parthenium hysterapharus L.)

plant residue on growth of salvinia and paddy seedlings was studied. The
inhibitory activity of the residue as shown by its effect on the number of
healthy fronds (HFN) and biomass was in the order: flower and leaf > stem
and root. The flower (FP) and leaf (LP) residue was lethal at and above 0.75%
(w/v, the convention used throughout), and inhibitory at lower doses. The
stem (SP) and root (RP) residue supported growth of salvinia at lower doses
and were slightly inhibitory at higher (I.25%) dose. All the above residue
types supported the growth of paddy seedlings except at 1.25%, the highest
concentration tested, which was slightly inhibitory. The amounts of chloro-
phyll a, b, total chlorophyll, and carotenoid pigments in the leaves of the
paddy seedlings grown in the medium were comparable to the amounts in the
leaves of seedlings grown in distilled water. This demonstrates beneficial
effects of the treatments. The study shows that salvinia is more sensitive to
allelochemicals released by FP and LP into the aqueous medium. Both salvinia
and paddy responded similarly to SP and RP by supporting growth at lower
doses, probably due to lower levels of inhibitors. The results are discussed
with reference to the possible role of allelopathy by parthenium on the pop-
ulation dynamics of aquatic weeds in natural ecosystems.

Key Words--Parthenium hysterophorus, Salvinia molesta, O~za sativa, bio-

mass, inhibitory activity, chlorophyll, carotenoid.

INTRODUCTION

Parthenium (Parthenium hysterophorus L.) is an a n n u a l w e e d e n d e m i c to the

A m e r i c a s that h a s s p r e a d to A f r i c a , A u s t r a l i a , a n d A s i a ( T o w e r s et al., 1977).
T h e w e e d has b e c o m e a s e r i o u s p r o b l e m f o r a g r i c u l t u r e and the e n v i r o n m e n t .

3123

(KlN8-O331/94/1200-3123~OT.(K)/(J ~c, 1994 Plenum Publishing C*~rlx~ratit~n

3124 PANDEY

Sesquiterpene lactones and phenolics form the main water-soluble inhibitory

allelochemicals in the plant's parts (Kanchan and Jayachandra, 1980). Parthen-
ium leaves and inflorescence contain the highest quantity of parthenin, followed
by the stem and roots, successively, while total phenolics were highest in leaves
followed by inflorescence, roots, and stem, successively (Kanchan and Jaya-
chandra, 1980). Investigations of the inhibitory effect of parthenium residue on
water hyacinth (Pandey et al., 1993a,b) showed that the leaf and flower powder
were lethal at relatively lower concentration, and stem residue was lethal at a
double dose, killing the treated plants in about a month. Root residue was
inhibitory at a relatively much higher dose. On the other hand, the stem and
root residue at lower doses supported growth of the treated plants, probably due
to lower levels of inhibitors, allowing utilization of nutrients from the residue.
It was observed that in aquaculture at different levels of parthenium plant resi-
due, water hyacinth plants were much more sensitive to inhibitory activity,
implying that water hyacinth could possibly be controlled by using a strongly
allelopathic terrestrial weed. Studies have confirmed that parthenium leaf residue
is more toxic to aquatic weeds--water hyacinth and salvinia--than to wheat
seedlings (Pandey et al., 1993a: Pandey, 1994). The present investigation was
undertaken to study the relative effects of the residue of different parts of par-
thenium on salvinia plants and paddy seedlings with reference to its possible
use in the management of salvinia in paddy fields.

METHODS AND MATERIALS

Salvinia Plants. The plants maintained in culture as described earlier (Pan-

dey, 1994) were used for these experiments.
Parthenium Plant Part Residue. The flowers, leaves, stem, and roots were
collected from plants growing near the National Research Centre for Weed
Science, Jabalpur (M.P.), India, were powdered to pass through an 80-mesh
sieve, and stored until use following the procedure described in detail in Pandey
et al. (1993b).
Effect of Residue of Plant Parts of Fronds and Biomass. Residues of par-
thenium flower (FP), leaf (LP), stem (SP), and root (RP) were each dispersed
in 20 liters of tap water in plastic tubs to make 0.25%, 0.50%, 0.75%, 1.00%,
and 1.25% (w/v, the convention used throughout) solutions, following the
method described earlier (Pandey et al., 1993a). Tap water was used since a
large quantity was required. Ten preweighed salvinia plants, each consisting of
20 fronds, were placed on the water in the tubs and were allowed to grow under
natural outdoor conditions during 1992-1993. Water lost due to evapotranspir-
ation was replenished regularly to keep the volume of water in the tubs constant.
Numbers of healthy fronds and biomass were monitored throughout the period
INHIBITION OF SALVtNIA BY PARTHENIUM 3125

of the experiments up to 90 days. A frond not showing wilting or drying from

the marigns or desiccation was considered to be healthy.
Analysis of Phenolic Acids in Medium. The medium was sampled 72 hr
after suspending the plant residue in water filtered through Whatman No. 1 filter
paper, and the phenolic acids were determined using the method of Swain and
Hillis (1959).
Effect of Parthenium Residue on Growth and Pigments in Paddy. Paddy
(Oryza sativa L. var. Kranti) seeds were germinated over wet Whatman No. 1
filter paper at 25C following the method of the International Seed Testing
Association (Anonymous, 1985), Twenty seedlings, each with a 2-cm radicle,
were placed on a perforated aluminum foil cover with the radicle protruding
down into 100 ml of a selected medium containing FP, LP, SP, RP, water
(control), or a quarter strength nutrient medium (Jain et al., 1989) in 100-ml
beakers. The seedlings were allowed to grow for 10 days during which evapo-
transpiratory loss of liquid was replenished by adding water regularly. There-
after, the seedlings were removed, washed with water, and root and shoot lengths
and fresh weights were measured. Leaf chlorophyll a, b, and total chlorophyll
were determined using the procedure of Arnon (1949). Carotenoids were assayed
using the procedure described by Canal et al. (1985).
All experiments were repeated three times. The data were subjected to
ANOVA and differences of means were estimated by calculating least significant
differences. Values with a lethal dose showing 100% response were excluded
from statistical analysis.

RESULTS AND DISCUSSION

The results (Figures 1-4) show that different parts of parthenium varied in
their inhibitory activity on salvinia plants. The FP at 0.25% was stimulatory to
the HFN at five or 10 days (Figure la). Subsequently it was inhibitory. The FP
at 0.25% and 0.50% was inhibitory to the biomass, but apparently not at a
significant level until 10 days (Figure lb). The symptoms--dull green color,
desiccation, browning, and finally initiation of blackening from older fronds
were evident. Inhibition depended on the concentration of the residue. At and
above 0.75%, FP was lethal, killing the salvinia plants. FP (Figure 2a,b) was
similarly inhibitory at lower levels and was lethal at and above 0.75%. The SP
and RP were not lethal even at 1.25%, the highest concentration tested. Lower
levels of SP caused little enhancement in HFN, but proved to be inhibitory at
higher concentrations. The inhibitory activity was highest at 1.25 %. The SP at
lower levels increased biomass over the plants grown in water. The RP did not
inhibit HFN at the lowest concentration (0.25%) until 15 days; by contrast it
increased the biomass of the treated plants. At and above 0.50% the biomass
3126 PANDEY

80
40

~_-3o
..~----F--.~fm ---'----Y T
05 IO 15 20 Z5 80
Tre~t~nt durati0n In davs

~ ~ NFP o,25~
_ ~ 8o I F P [Link]
4Q [] FP 0,75%

05 I0 15 Z0 Z5 80
Treatmnt durationin days

FIG. 1. Effect of parthenium flower residue (FP) on (a) number of healthy fronds (HFN)
and (b) biomass of salvinia.

=zol ~ B '" " ~'~Control

[ ] L P o,zsx
ILP o,5~
~LP o,Tsx

!~LP 1,25X
ILSD ~t 5x
I I i i i
O5 iO 15 20 25 ;0
T r e ~ t duration in d~s

FIG. 2. Effect of parthenium leaf residue (LP) on (a) number of healthy fronds (HFN)
and (b) biomass of salvinia.
INHIBITION OF SALVINIA BY PARTHENIUM 3127

180

120

~-~0
05 10 15 20 25 30

Z40I
ZIO ~sp o,25~
1801
.~ ~50' IsP o,~ox
r'/'~SP 0,75}{
~SP I,O0'X
~sP ~,zsx
.. 5~

10 I5 20 25 30
Treatmentd~r~lionin da~s

F~G. 3. Effect of parthenium stem residue (SP) on (a) number of healthy fronds (HFN)
and (b) biomass of salvinia.

60 ~I
e , i m

I I I I i

180

120
~RP hCq~
~'~o~o ERP ~.zss
~' 20 ILSD at 5~
0 05 i I0 ~ 15 ~ 20 ~ 25 ~ 30
Trealmnt dvaH0n in days

Fi& 4. Effect of parthenium root residue (RP) on (a) number of healthy fronds (HFN)
and (b) biomass of salvinia.
3128 PANDEY

increase was reduced considerably. Thus, the SP and RP at lower levels increased
biomass over the plants grown in water. The high values of LSD obtained were
probably due to elimination of the data concerning the lethal dose, which was
at t00% in all cases and due to wide variation in the response of salvinia plants
to parthenium residue below lethal doses.
Phenolic acids at 1% residue were maximum in the medium containing FP
(180 ppm), following by LP (138 ppm), RP (85 ppm), and SP (62 ppm).
Phenolic acid yield percent of residue was maximum at 1.80 _+ 0.10 in FP,
followed by 1.38 + 0.08 in LP, 0.85 + 0.02 in RP, and 0.62 + 0.05 in SP.
The inhibitory activity of the parthenium plant parts residue to salvinia is in the
order: FP and LP > SP and RP. This coincides with the relatively higher levels
of phenolic acids from FP and LP than SP and RP. Parthenium leaves and
inflorescence have been found to contain the highest quantity of parthenin tbl-
lowed by the stem and roots (Kanchan and Jayachandra, 1980). The much more
inhibitory activity of FP and LP than SP and RP to salvinia is probably due to
the higher levels of the allelochemicals, including phenolic acids and parthenin,
in the residue. Growth of paddy seedlings, as revealed by root and shoot lengths
and fresh weights, was enhanced by parthenium plant residue at lower levels
and inhibited at higher levels when compared with growth of plants in water
(Figures 5 and 6). In some cases, the treated plants showed higher growth than
those grown in the nutrient medium. This confirms the beneficial effects of the
parthenium residue at lower levels, but at higher levels the growth increment

16
14

8
~6

~Z
0 i I i

36
[]C0nlr01
~ 3O

~. 24
= IB

~ t2

IkSD at 5X
I i I
FP LP SP RP
Tre~t~nt

Fie. 5. Effect of parthenium plant part residue on (a) seminal root length and (b) fresh
weight of paddy var. Kranti seedlings after 10 days of growth in an aquaculture test for
inhibitory activity.
INHIBITION OF SALVINIA BY PARTHENIUM 3129

was inhibited and showed growth reduction over the plants grown in water.
Leaves of seedlings grown in medium containing [Link] residue were a
very lush green in color. The beneficial effect was also evident from enhanced
chlorophyll a, b, total chlorophyll, and carotenoids in the leaves of seedlings
grown in medium containing LP (Figure 7). Although the seedling growth at
higher levels of LP was comparable to or a little lower than those grown in
water, the pigments were considerably higher in the leaves.
The results show that at a particular concentration, FP and LP had the
highest inhibitory activity due to higher levels of inhibitory allelochemicals
leaching out of the residue into the medium. While FP and LP were inhibitory

16

8 l0

--6
g

2
0 i i I

=~36

o , , , {IILSD~t s~ t
fP LP SP ,~P
Tr.~at n'~.nt

FIG. 6. Effect of parthenium plant part residue on (a) shoot length till margin of the
longest leaf and (b) fresh weight of paddy var. Kranti seedlings after 10 days of growth
in an aquaculture test for inhibitory activity.

!i f;! ~-': [] o,7~

~; 2O ,'. "- ~ ,7,~." ) IILSD at 51

Chl a Chl b Total Chl C~ole~olds

FIe. 7. Effect of parthenium leaf residue (LP) on chlorophyll a, b, total chlorophyll,

and carotenoids in the leaves of paddy var. Kranti seedlings after 10 days of growth in
aquaculture,
3130 PANDEY

for salvinia at lower doses and lethal at higher doses, SP and RP improved
growth of the treated plants at lower levels and proved to be slightly inhibitory
at higher levels. By contrast, the paddy seedlings responded to FP, LP, SP, and
RP more or less similarly, as lower doses usually improved the growth and
higher doses were marginally inhibitory. The improvement in growth over the
seedlings grown in water was, at certain levels, equal to or more than that of
the seedlings grown in nutrient solution.
The FP and LP have relatively more of sesquiterpene lactones and phenolic
acids. It is evident that the salvinia plants are much more susceptible to the
inhibitory activity of these allelochemicals than paddy seedlings. Interestingly,
the allelochemicals present in SP and RP appear almost equally inhibitory to
both salvinia and paddy seedlings. This might be due to different--or differences
in the composition of--allelochemicals, or both. Promotion of growth of salvinia
plants at lower doses of SP and RP and of paddy seedlings by FP, LP, SP, and
RP might be due to utilization of nutrients from the residue. This was probably
due to a low level of allelochemicals, which was unable to inhibit growth of
the treated plants. The beneficial effect of parthenium residue on paddy was
consistent in pot experiments (data not presented),
Consistent with earlier findings (Pandey et al., 1993a,b), in a study (Pan-
dey, unpublished), parthenium FP and LP at 0.25-0.50% and SP and RP at or
above 1.00% were found lethal to other aquatic weeds such as Pistia stratiotes
L., Azolla nilotica Decne., Lemna pausicostata Hegelm., Spirodella polyrrhiza
(L.) Schleid., and to all submerged aquatic weeds, such as Hydrilla verticitlata
L.f. Royle, Ceratophylum demersum L., and Najas gramineae Del. Thus, it is
established that parthenium plant residue is relatively more toxic to aquatic
weeds than to test crop species (wheat and paddy). The implications of accu-
mulation of parthenium plant residue in the environment on the population
dynamics and shifts in flora of aquatic weeds in natural ecosystems need carefid
consideration, as accumulation of enormous quantities of the residue under nat-
ural conditions appear to be speculative for the following two reasons: (1) Pic-
man and Picman (1984) reported that sesquiterpene lactones, parthenin and
traces of coronopitin, which are found in large quantities in parthenium are toxic
to its seedlings and older plants at a concentration of 0.1%. Obviously, the
water-soluble plant metabolites have been suggested to play important roles in
allelopathy and defense against herbivorous predators and diseases, as autotoxins
in the regulation of populations, and in the timing of the germination process
(Picman and Picman, 1984). (2) The toxicity of parthenium residue in the aquatic
environment declines at an appreciable rate as the lethal dose of LP and FP
have been found to support growth of salvinia plants freshly placed in the
medium under outdoor conditions for about a month (Pandey, unpublished data).
Thus, even if considerable amounts of inhibitors find their way or are washed
by rain into waterbodies, building up of inhibitors to such an extent as to make
INHIBITION OF SALVINIA BY PARTHENIUM 3 13 1

d e t e c t a b l e c h a n g e s in t h e o c c u r r e n c e a n d p o p u l a t i o n d y n a m i c s o f s a l v i n i a a n d
o f o t h e r a q u a t i c w e e d s is u n l i k e l y . H o w e v e r , p a r t h e n i u m r e s i d u e c o u l d be
e x p l o r e d as a p o s s i b l e s t i m u l a n t f o r p a d d y s e e d l i n g g r o w t h a n d a s a n a t u r a l
p e s t i c i d e in p a d d y fields f o r t h e m a n a g e m e n t o f s o m e a q u a t i c w e e d s . T h e f i n d i n g
i n d i c a t e s t h e p o s s i b i l i t y o f u s e o f a s t r o n g l y a l l e l o p a t h i c terrestrial w e e d f o r
b i o l o g i c a l c o n t r o l o f w e e d s like s a l v i n i a a n d w a t e r h y a c i n t h a n d o t h e r f l o a t i n g
and submerged aquatic weeds.

Acknowledgments--The author thanks Mr. S. Dhagat for helping with illustrations, and Dr.
M.A.K. Lodhi for his critical review of an earlier copy of the manuscript.

REFERENCES

ANONYMOUS. 1985. International rules for seed testing. International Seed Testing Association. Seed
Sei. TeehnoL 13:356-513.
ARNON, D.I. 1949. Copper enzymes in isolated chloroplast. Polyphenol oxidase in Beta vulgaris.
Plant Physiol. 24: 1-15.
CANAL V~LLANUEVA,M.J., MLINIZ, B.F,, and TAMES, R.S. 1985. Effects of glyphosate on growth
and the chlorophyfl and carotenoid levels of yellow nutsedge (Cyperas esculentus). Weed ScL
33:751-754.
J;,IN, R.. SINGH, M~, and DEZMAN, D.J. 1989. Qualitative and quantitative characterization of
phenolic compounds from lantana (Lantana camara) leaves, Weed Sci 37:302-307.
KANCHAN, S.D., and JAYACHANDRA. 1980. Allelopathic effects of Parthenhtm hysterophoras L.
IV, Identification of inhibitors. Plant Soil 55:67-75.
PANOEY, D.K. t994, Inhibition of salvinia (Salvinia molesta Mitchell) by parthenium (Parthenium
hysterophorus L,). 1. Effect of leaf residue and allelochemicals. J. Chem. EeoL 20:000-000.
PANDEY, D.K., KAURAW,L.P., and BHAN, V.M. 1993a. Inhibitory effect of Partheniam (Parthen-
ium hysterophorus L.) residue on growth of water hyacinth (Eiehhornia erassipes Mart Solms.)
I. Effect of leaf residue. J. Chem. Eeol. 19:2651-2662.
PANDEY, D.K.. KAURAW,L.P., and BHAN, V.M. 1993b. Inhibitory effect of parthenium (Parthen-
iunl hysterophoras L,) residue on growth of water hyacinth (Eiehhornia erassipes Mart Solms.)
11. Relative effect of flower, leaf, stem, and root residue. J. Chem. EcoL 19:2663-2670.
PICMAN, J., and PICMAN, A.K. 1984. Autotoxicity in Parthenium hysterophorus and its possible
role in control of germination. Biochem. Syst. Ecol. 12:287-292.
SWAIN, T., and HILLIS, W.E. 1959. The phenolic constituents of Prunus &~mestica. I. The quan-
titative analysis of phenolic constituents. J. Sci. Food Agric. 10:63-68.
TOWERS, G H . N . , MITCHELL,J.C., RODRIGUEZ,E., BENNETT, F.D,, and SUBBARAO,P.V. 1977.
Biology and chemistry of Partheniurn hysterophorus L., a problem weed in India. J. &~i. h~d.
Res, 36:672-684.