Journal of Eco-friendly Agriculture 19(1): 175-178. 2024 U. H. Bhimani and N. M.

Gohel ISSN : 2229-628X (print)

doi: 10.48165/jefa.2024.19.01.29 e-ISSN : 2582-2683 (online)

Research Article

Management of Alternaria leaf spot of ashwagandha through

eco-friendly inputs
U.H. Bhimani* and N.M. Gohel
Department of Plant Pathology, B. A. College of Agriculture, Anand Agricultural University, Anand-388 110, Gujarat, India.
*e-mail: urjabhimani@[Link]
ABSTRACT

The Alternaria leaf spot disease of ashwagandha is a significant and concerning issue, posing a major threat to
the crop’s health and productivity. The use of eco-friendly inputs provides an approach to effectively managing
disease and contributes to the reduction of environmental pollution. Six organic inputs were tested under in vitro
conditions against Alternaria alternata. Panchagavya was found effective in inhibiting the pathogen which showed 89.28
per cent growth inhibition. Among the six treatments tested under field conditions, the foliar application of three
sprays of neem leaf extract, 10 per cent (1 l 10 l -1 of water) at 15 days intervals recorded minimum per cent disease
intensity (22.93%) with maximum yield (668 kg ha-1 ), which was statistically at par with foliar spray of azadirachtin
(0.03% EC, 0.00015%).
Key words: Alternaria alternata, ashwagandha, eco-friendly.

Ashwagandha (Withania somnifera), also known as growth, yield, and medicinal properties. In addition to the
Indian ginseng or winter cherry, is an ancient medicinal economic impact, plant diseases can also affect the chemical
herb with a rich history of traditional use in Ayurvedic composition and bioactive compounds present in
medicine. The entire plant as such possess important ashwagandha. Among the diseases, leaf spot caused by
medicinal value, however, roots of W. somnifera are a rich Alternaria alternata (Mishra et al., 2018; Meena et al., 2019;
source of bioactive compounds which include alkaloids and Rahman et al., 2019) is a major concern and adversely affects
secondary metabolites such as withaferin-A, withanolides, commercial cultivation as well as the production of the crop.
sterols and phenols etc. (Ghosal et al., 1988; Kandil et al., The attack of pathogen not only reduces the chlorophyll and
1994; Bhattacharya et al., 2002). It has been estimated that reducing sugar contents but also causes a reduction in
the alkaloid content of Indian roots ranges from 0.13 to 0.31 withaferin- A and withanolides contents by 15.4 and 76.3
per cent. In India, it is successfully grown in Madhya per cent, respectively (Pati et al., 2008) that confer immense
Pradesh, Gujarat, Uttar Pradesh, Haryana, Rajasthan and therapeutic value to the crop.
Punjab (Bhatia et al., 1987) for root yields. Ashwagandha is
Furthermore, the plant parts are used directly as
found wild in grazing grounds in Mandsaur district of
medicine. Uses of chemical fungicides are extremely harmful
Madhya Pradesh and the forest lands in the Bastar district
to humans. Replacement of chemical fungicides with
of Chhattisgarh, all over the foothills of Punjab, Himachal
biocontrol agents and organic inputs is an alternative means
Pradesh and Western Uttar Pradesh. In India, the crop is
to manage plant pathogens, produce safe food and reduce
grown over an area of around 4000 ha, primarily in the drier
environmental pollution. Hence, there is a vital need for
areas of Punjab, Sindh, and Rajasthan as well as Manasa,
effective and ecologically safe management practices to be
Neemach, and Jawad tehsils in Mandsaur district of Madhya
developed that do not hamper the medicinal value of the
Pradesh (Rajamani et al., 2019). The expected production of
crop.
its roots in India is more than 1500 tonnes, while the annual
requirement is over 7000 tonnes (Baghel et al., 2010). MATERIALS AND METHODS
Therefore, it is needed to increase the cultivation of The fungal pathogen causing Alternaria leaf spot
ashwagandha for higher production. disease was isolated from infected ashwagandha leaves from
However, the cultivation of ashwagandha is often a Research Farm, Department of Plant Pathology, BACA,
hindered by various diseases that can profoundly impact its AAU, Anand (Fig. 1). The fungus was isolated from diseased

©2024 of Eco-friendly Agriculture 19(1) 2024

Journal 175
doi: 10.48165/jefa.2024.19.01.29
 Management of Alternaria leaf spot of ashwagandha through eco-friendly inputs

tissues using standard methodology on potato dextrose agar Table 1: Disease rating scale against Alternaria leaf spot in
(PDA) (Tuite, 1969) amended with streptomycin sulphate. ashwagandha.
After isolation, the pathogen was incubated in BOD at 28±1 Rating Scale Description
°C and the culture obtained was purified by hyphal tip 0 Leaves free from infection or 0 to 5% leaf area with lesions
method (Rangaswami, 1972). Small irregular brown spots with concentric rings
1
covering 6-10 per cent leaf area
Evaluation of organic inputs against A. alternata under in- Lesions enlarging, irregular brown with concentric rings
2
vitro conditions covering 11-25 per cent of leaf area
Lesions coalesce to form irregularly and appear as a
Six organic inputs were evaluated against A. alternata 3
typical leaf spot covering 26-50 per cent of the leaf area
under in vitro conditions. All the organic inputs were tested Lesions coalesce to form irregularly and appear as a
at three different concentrations, i.e., 5, 7 and 10 per cent 4
typical blight symptom covering >50 per cent leaf area
following the poisoned food technique (Grover and Moore,
1962). Two replications were maintained for each treatment. Table 2: Treatment details.
Required quantities of organic inputs were thoroughly mixed
Tr. No. Treatments
in PDA before pouring in 90 mm sterilized petri plates and
Foliar spray of T. viride (1% WP, 2x109 cfu g-1), 50 g 10 litre-1 of
allowed to solidify. Simultaneously, without any treatment, T1
water
PDA was poured into petri plates as a control. The plates Foliar spray of P. fluorescens (1% WP, 1x108 cfu g-1), 50 g 10 litre-
T2
were incubated at 28±1 °C. The colony diameters were 1 of water

measured after 10 days when the control plates were full of T3

Foliar spray of neem leaf extract, 10 per cent (1 litre 10 litre-1 of
fungal growth. Per cent growth inhibition over control was water)
calculated by using the formula given by Vincent (1947). Foliar spray of azadirachtin, 0.03% EC, 0.00015% (50 ml 10 litre-
T4 1 of water)

DC − DT Foliar spray of cow urine (desi), 5 per cent (500 ml 10 litre-1 of

T5
PGI = × 100 water)
DC T6 Foliar spray of Panchagavya, 5 per cent (500 ml 10 litre-1 of water)
T7 Control (Untreated check)
PGI = Per cent Growth Inhibition
DC = Colony diameter in control (mm) PDI in control − PDI in treatment
Disease control(%) = × 100
DT = Colony diameter in treatment (mm) PDI in control

Management of Alternaria leaf spot of ashwagandha under RESULTS AND DISCUSSION

field conditions through eco-friendly inputs Evaluation of organic inputs against A. alternata under in-
The field experiment was conducted in rabi 2021-22 vitro conditions
using the variety Gujarat Anand Ashwagandha 1 at A total of six organic inputs with three different
Research Farm, Department of Plant Pathology, BACA, AAU, concentrations (5, 7 and 10%) were evaluated in-vitro by the
Anand. Six treatments were used for the eco-friendly poisoned food technique. Data presented in Table 3 showed
management of Alternaria leaf spot. The experiment that at 10 per cent concentration all organic inputs were
comprised seven treatments with three replications in a capable of inhibiting the mycelial growth of A. alternata as
randomised block design (Table 2). The first foliar spray of compared to control. Panchagavya inhibited the highest
the respective treatment was given, commencing from the
appearance of the disease and a subsequent spray was given
after 15 days of the first spray. The disease intensity was
recorded after seven days of each spray. Ten plants were
selected randomly from each plot and per cent disease
intensity was worked out. PDI and the per cent disease
control were calculated by using the formula given by
Wheeler (1969). Disease intensity was calculated using a
scale of 0-4 as described by Meena et al. (2019) in Table 1. Dry
root yield was recorded plot-wise and at the time of harvesting
and finally converted into kg per ha.
Figure 1. Symptoms of Alternaria leaf spot disease of
ashwagandha.

176 Journal of Eco-friendly Agriculture 19(1) 2024

doi: 10.48165/jefa.2024.19.01.29
 U. H. Bhimani and N. M. Gohel

Table 3: Effect of organic inputs on mycelial growth Table 4: Effect of eco-friendly inputs on Alternaria leaf spot
inhibition of A. alternata under in vitro conditions. of ashwagandha under field conditions.
Tr. Organic Concentration Mycelial Growth Disease intensity (%) Disease Yield
Dry
No. inputs (%) growth (mm) inhibition (%) After After After reduction increase over
Tr. No. root
T1 5 76.55c 14.94 1st 2nd 3rd over control
yield
T2 Cow urine 7 70.30d 21.89 spray spray spray control (%) (%)
(Fresh) 18.84cd 26.70c 34.32cd
T3 10 64.60e 28.22 T1 29.78 585abc 17.94
(10.50) (20.30) (31.83)
T4 5 65.00e 27.78 21.77b 32.56b 37.38bc
T2 18.62 537ab 8.27
T5 Cow dung 7 63.70ef 29.22 (13.80) (29.02) (36.89)
T6 10 63.00ef 30.00 15.22 19.36
e e 22.93 f
T3 66.18 668c 34.68
T7 5 70.34d 21.84 (7.00) (11.12) (15.33)
T8 Buttermilk 7 67.24de 25.29 17.37de 23.38d 27.56e
T4 52.39 604bc 21.77
T9 10 66.28de 26.36 (9.07) (15.85) (21.58)
T10 5 44.68h 50.36 21.60b 33.55b 39.02b
T5 12.49 521ab 5.04
T11 Panchagavya 7 14.37i 84.03 (13.60) (30.55) (39.67)
T12 10 9.65j 89.28 19.52 29.11 33.74
c c d
T6 31.79 557ab 12.30
T13 5 60.10f 33.22 (11.28) (23.72) (30.92)
T14 Jeevamrutha 7 55.00g 38.89 24.65a 36.13a 42.31a
T7 - 496a -
T15 10 53.00g 41.11 (17.43) (34.80) (45.33)
T16 5 86.00ab 4.44 [Link].± 1.08 1.38 1.78 - 27.87 -
T17 Beejamrutha 7 85.00b 5.56 C.D. at 5% Sig. Sig. Sig. - Sig. -
T18 10 84.00b 6.67 C.V. (%) 9.41 8.33 9.11 - 8.52 -
T19 Control - 90.00a - Treatment means with the letter/letters in common are not
[Link].± - 1.32 - significant by Duncan’s new multiple range test (DNMRT) at a
C.D. at 5% - Sig. - 5% level of significance.
C.V. (%) - 2.99 -
Management of Alternaria leaf spot of ashwagandha under
Treatment means with the letter/letters in common are not
field conditions through eco-friendly inputs
significant by Duncan’s new multiple range test (DNMRT) at a 5
% level of significance. The data presented in Table 4 revealed that the lowest
disease intensity (22.93%) and highest disease control
(66.18%) were recorded in treatment T3, i.e., foliar spray of
mycelial growth of 50.36, 84.03 and 89.28 per cent. The next
neem leaf extract (1 l 10 l-1 of water) (Fig. 2). The next best
better organic input Jeevamrutha inhibited 41.11 per cent
treatment was T4 with disease intensity (27.56%) and disease
mycelial growth at 10 per cent concentration. Similarly,
control (52.39%). The highest disease intensity (39.02%) was
Pandia et al. (2019) found that Jeevamrutha was most effective
witnessed in treatment of foliar spray of cow urine (desi),
in inhibiting A. alternata followed by cow urine at 7.5 per
(T5) with lower disease control (12.49%). However, the utmost
cent concentration.

Control (Untreated check)

Figure 2. Foliar spray of neem leaf extract, 10 per cent (1 l 10 l-1 of Figure 3. Eco-friendly management of Alternaria leaf spot of
water). ashwagandha under field conditions.

Journal of Eco-friendly Agriculture 19(1) 2024 177

doi: 10.48165/jefa.2024.19.01.29
 Management of Alternaria leaf spot of ashwagandha through eco-friendly inputs

disease intensity of 42.31 per cent was recorded in the fungicides against brown rot organism, Sclerotinia fruticola
untreated check (Fig. 3). Hassanein et al. (2010) and Murmu and S. laxa. Phytopathology, 52: 876-880.
et al. (2015) found that foliar application of neem leaf extract Hassanein, N.M., Zeid, M.A.A., Youssef, K.A. and Mahmoud,
provided better control against the early blight (A. solani) in D.A. 2010. Control of tomato early blight and wilt using
tomato and early blight (A. solani) in potatoes. Similarly, aqueous extract of neem leaves. Phytopathologia Mediterranea,
49: 143-151.
Vibha and Upadhyay (2016) reported that Azadirachtin
Kandil, F.E., El Sayed, N.H., Abou-Douh, A.M., Ishak, M.S. and
(0.15%) × T. asperellum @ 106 cfu ml-1 + P. fluorescens 21 @ 109
Mabry, T.J. 1994. Flavonol glycosides and phenolics from
cfu ml-1 was the best treatment for reducing the Alternaria Withania somnifera. Phytochemistry, 37: 1215-1216.
leaf blight in ashwagandha and enhancing the root and seed Meena, R.P., Kalariya, K.A., Saran, P.L. and Manivel, P. 2019.
yield. Mishra (2021) remarked that P. fluorescens @ 0.5 per Evaluation of ashwagandha (Withania somnifera L.) Dunal
cent, recorded the highest disease control followed by accessions and breeding lines against leaf spot disease
Trichoderma harzianum and Panchagavya. caused by Alternaria alternata under subtropical condition
of India. Journal of Applied Research on Medicinal and Aromatic
The significantly highest dry root yield was recorded Plants. [Link]
in T3 (668 kg ha-1) which was statistically at par with T4 (604 Mishra, A., Singh, S.P., Mahfooz, S., Singh, S.P., Bhattacharya, A.,
kg ha-1) as compared to control. Whereas, the maximum yield Mishra, N. and Nautiyal, C.S. 2018. Endophyte-mediated
increase over control was obtained in T3 (34.68%). modulation of defence-related genes and systemic
resistance in Withania somnifera (L.) Dunal under Alternaria
CONCLUSION alternata stress. Applied Environmental Microbiology, 84: 1-13.
Panchagavya was found most effective under in vitro Mishra, R.S. 2021. Management of Alternaria leaf spot of
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ashwagandha. The treatment recorded the lowest mycelial The Pharma Innovation, 10: 1270-1272.
growth (9.65 mm) and maximum growth inhibition (89.28%) Murmu, S., Dey, S. and Chakraborty, A. 2015. Management of
at a 10 per cent concentration. Under field conditions, foliar early blight of potato using biocontrol agents and plant
spray of neem leaf extract (1 l 10 l-1 of water) was found most extracts. Journal of Applied and Natural Science, 7: 860-864.
effective with the lowest disease intensity (22.93%) and Pandia, S., Trivedi, A., Sharma, S.K. and Yadav, S. 2019. Evaluation
maximum disease reduction over control (66.18%) against of Jeevamrut and its constituents against Alternaria leaf spot
of mungbean in-vitro and under cage house condition in
Alternaria leaf spot of ashwagandha followed by foliar spray
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ACKNOWLEDGEMENTS Pati, P.K., Sharma, M., Salar, R.K., Sharma, A., Gupta, A.P. and
Singh, B. 2008. Studies on leaf spot disease of Withania
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Department of Plant Pathology, B. A. College of Agriculture, Journal of Microbiology, 48: 432-437.
Anand Agricultural University, Anand for providing all the Rahman, M.A., Moni, Z.R., Hussen, M.A. and Ansari, T.H. 2019.
facilities required for smooth conduct of experiments. Prevalence of fungal diseases in medicinal plants in
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178 Journal of Eco-friendly Agriculture 19(1) 2024

doi: 10.48165/jefa.2024.19.01.29