In vitro efficacy of bioagents and fungicides against chickpea wilt caused by Fusarium oxysporium f. sp. ciceri

Introduction

Chickpea (Cicer arietinum L.) is one of the most important pulse crops grown and consumed worldwide, particularly in Afro-Asian countries. In India, it is a major rabi pulse cultivated under both rainfed and irrigated conditions. Chickpea grains contain about 21.1% protein, 61.5% carbohydrates and 4.55% fat, with protein quality superior to many other pulses. During 2019-2020, chickpea occupied 9.85 million ha in India with a total production of 10.90 million tonnes and an average productivity of 1142 kg ha⁻¹. It is cultivated in 16 states, with Madhya Pradesh, Rajasthan, Uttar Pradesh, Maharashtra and Haryana as the leading producers. Madhya Pradesh tops the list with 26.8 lakh ha, whereas Maharashtra ranks second with 25.94 lakh ha and a production of 28.46 lakh tonnes at 1097 kg ha⁻¹ productivity (Chief Statistician, Maharashtra, 2021). Due to its nitrogen-fixing ability, chickpea improves soil fertility and can restore up to 140 kg N ha⁻¹ during its growth period (Poonia and Pithia, 2013) [12] . Despite its importance, chickpea is susceptible to more than 50 pathogens (Nene et al., 1981) [10] . Wilt (Fusarium oxysporum f. sp. ciceri), root rot (Rhizoctonia bataticola), and collar rot (Sclerotium rolfsii) are major constraints in Marathwada (Maharashtra) and many other chickpea-growing regions. Wilt disease occurs widely across India, Pakistan, Iran, Nepal, Myanmar, Mexico, Spain, Peru, Syria and the USA, causing regular yield losses of 5-10%, which may reach 60-70% during severe epidemics (Singh and Dahiya, 1973) [15] . The pathogen F. oxysporum f. sp. ciceri was first reported in India by Butler (1918) [4] . It is a facultative, soil-borne saprophyte that can persist as chlamydospores or mycelium for up to six years even in the absence of a susceptible host (Haware et al., 1986a) [7] . The fungus colonizes the xylem vessels, blocks water translocation, and leads to rapid wilting and plant death (Padwick, 1948; De et al., 1996) [11,5] . Several approaches have been investigated to manage wilt, including resistant varieties (Brisa et al., 2007) [3] , chemical control, plant extracts (Kumar and Tripathi, 1991) [8] and biological control agents (Rojo et al., 2007; Dubey et al., 2007) [14,6] . Given the seriousness of the disease and the significant yield losses it causes, the present investigation was undertaken to develop an effective integrated management strategy for chickpea wilt.

Materials and Methods

An experiment for the management of wilt of chickpea caused by Fusarium oxysporum f. sp. ciceri using different fungicides and bioagents was carried out at Department of Plant Pathology, College of Agriculture, Badnapur, during 2024-25.

In vitro efficacy of bioagents

A total of seven fungal bioagents as detailed under treatments were evaluated in in vitro against Fusarium oxysporum f. sp. ciceri, by applying Dual culture technique (Arora and Upadhay, 1978) [1] . Seven days old pure cultures of the test pathogen and test bioagents grown on PDA medium were used for the study. Two 5 mm culture discs, one each of the test pathogen and the test bioagent were cut out with sterilized cork borer and placed at equidistance and exactly opposite to each other on autoclaved and solidified PDA medium in Petri plates and were incubated at 25±1°C. PDA plates inoculated alone with pure culture disc (5 mm) of the test pathogen were maintained as control. Observations on radial mycelial growth and colony diameter of Fusarium oxysporum f. sp. ciceri was recorded when untreated control plates were fully covered with mycelial growth. Per cent mycelial growth inhibition of the pathogen with the bioagents, over untreated control was calculated by using formula suggested by (Arora and Upadhyay, 1978) [1] . In vitro efficacy of fungicides Efficacy of seven combi fungicides (each @ 1500, 1750 and 2000 ppm) was evaluated in vitro against Fusarium oxysporum f. sp. ciceri by applying poisoned food technique (Nene and Thapliyal, 1993) [9] . Three Petri plates / treatment / replication were maintained and also untreated control plates were maintained. Observations on radial mycelial growth / colony diameter of the Fusarium oxysporum f. sp. ciceri was recorded when untreated control plates were fully covered with mycelial growth. Per cent mycelial growth inhibition of the pathogen with the test fungicides over the untreated control were calculated by using the formula given by (Vincent, 1927).

Where, C = Growth of test pathogen in control T = Growth of test pathogen in treated plates

Detals of the experiment

Where, C = Growth of test pathogen in control T = Growth of test pathogen in treated plates.

Details of the experiment

Results and Discussion

Efficacy of bioagents against Fusarium oxysporum f. sp. ciceri The results reveled that, the colony diameter of mycelial growth and per cent inhibition of the F. oxysporum f. sp. ciceri with fungal antagonist ranged from 11.18 mm and 87.57% with T. harzianum to 48.12 mm and 46.53% with Bacillus subtilis.

Radial mycelial growth

Results (Table 1) revealed that among the seven bioagents tested, (T2) Trichoderma harzianum with 87.57 per cent mycelial growth was found most effective in controlling the mycelial growth of F. oxysporum f. sp. ciceri (11.18 mm) which was found significant over all of the treatments and (90.00 mm) untreated control (T8), (T6) Aspergillus niger was next best bioagent restricting mycelial growth of the F. oxysporum f. sp. ciceri (14.36 mm), followed by (T7) Pseudomonas fluorescens (22.27 mm), (T1) T. asperellum(28.44 mm), (T3) Verticillium lecanii (34.63 mm), (T5) Pink Pigmented Facultative Methylotrophs (PPFMs) (34.67 mm) and (T4) Bacillus subtilis (48.12 mm).

All the bioagents (Table 1) evaluated, exhibited fungistatic /antifungal activity against test pathogen and significantly inhibited its growth, over untreated control.

Inhibition of mycelial growth

Results (Table 1) revealed that out of the seven bioagents tested, (T2) Trichoderma harzianum was found most effective in inhibiting the mycelial growth of F. oxysporum f. sp. ciceri (87.57%) which was significant over rest of the treatments and untreated control (00.00%), The next best bioagent found in inhibiting the mycelial growth of the F. oxysporum f. sp. ciceri was (T6) Aspergillus niger (84.04%), which was followed by (T7) Pseudomonas fluorescens (75.25%), (T1) T. asperellum (68.40%), (T3) Verticillium lecanii (61.52%), (T5) Pink Pigmented Facultative Methylotrophs (PPFMs) (61.47%) and (T4) Bacillus subtilis (46.53%). The results are in agreement with the finding of several workers who reported that bioagents viz., Trichoderma harzianum, Aspergillus niger, Pseudomonas fluorescens, and T. asperellum had significantly inhibited mycelial growth of F. oxysporum f. sp. ciceri by Singh et al. (1977 and Sonawane and Pawar (2001) [16,17] .

Efficacy of fungicides against Fusarium oxysporum f. sp. Ciceri

Radial mycelial growth

The results (Table 2) revealed that at 1500 ppm concentration of the combi-product fungicides the radial mycelial growth of F. oxysporum f. sp. ciceri, ranged from 00.00 mm (T7) to 62.33 mm (T3). Minimum mycelial growth of F. oxysporum f. sp. ciceri was recorded in (T7) Carbendazim + Mancozeb (00.00 mm) which was followed by (T5) Carboxin + Thiram (12.28 mm). Whereas at 1750 ppm concentration of the combi-product fungicides the radial mycelial growth of F. oxysporum f. sp. ciceri, ranged from 00.00 mm (T7) to 43.79 mm (T3). Minimum mycelial growth of F. oxysporum f. sp. ciceri was recorded in (T7) Carbendazim + Mancozeb (00.00 mm) which was followed by (T5) Carboxin + Thiram (06.67 mm). Also, at 2000 ppm concentration of the combi-product fungicides the radial mycelial growth of F. oxysporum f. sp. ciceri, ranged from 00.00 mm (T7) to 31.22 mm (T3). Minimum mycelial growth of F. oxysporum f. sp. ciceri was recorded in (T7) Carbendazim + Mancozeb (00.00 mm) which was followed by (T5) Carboxin + Thiram (01.72 mm).

Inhibition of mycelial growth

The results (Table 2) revealed that at 1500 ppm concentration of the combi-product fungicides the per cent inhibition of mycelial growth of F. oxysporum f. sp. ciceri, ranged from 100.00 per cent (T7) to 30.74 per cent (T3). Maximum inhibition of F. oxysporum f. sp. ciceri was recorded in Treatment (T7) Carbendazim + Mancozeb with cent per cent (100.00%) inhibition. Whereas at 1750 ppm concentration of the combi-product fungicides the per cent inhibition of mycelial growth of F. oxysporum f. sp. ciceri, ranged from 100.00 per cent (T7) to 51.34 per cent (T3). Maximum inhibition of F. oxysporum f. sp. ciceri was recorded in Treatment (T7) Carbendazim + Mancozeb with cent per cent (100.00%) inhibition which was then followed by (T5) Carboxin + Thiram (92.58%). Also, at 2000 ppm concentration of the combi-product fungicides the per cent inhibition of mycelial growth of F. oxysporum f. sp. ciceri, ranged from 100.00 per cent (T7) to 65.31 per cent (T3). Maximum inhibition of F. oxysporum f. sp. ciceri was recorded in Treatment (T7) Carbendazim + Mancozeb with International Journal of Advanced Biochemistry Research https://www.biochemjournal.com cent per cent (100.00%) inhibition which was then followed by (T5) Carboxin + Thiram (98.01%).

The results in the present study were almost identical with previous researchers like Ravichandran and Hegde (2015) [13] who studied in vitro efficacy of contact, systemic and combi fungicides against pathogen causing chickpea wilt and observed that among these Carbendazim 12% + Mancozeb 63% (SAAF) was found effective at all concentrations (1000, 2000, 3000 ppm) with 100 per cent inhibition of Fusarium oxysporum f. sp. ciceri and least inhibition was recorded in Zineb 68% + Hexaconazole 4% WP (Avatar) 21.85% at 0.1% concentration.

References

1. Arora DK, Upadhyay R. Effect of fungal staling growth substances on colony interaction. Plant and Soil. 1978;49:685-690.
2. Biswas M, Ali SJ. Management of Fusarium wilt of chickpea (Cicer arietinum L.) under the undulating red and lateritic belt of West Bengal. Journal of Mycopathological Research. 2017;54(4):461-468.
3. Brisa R, Fernando MA, Asunción GS, Noemí MR, Arturo PE, José MD. The gene coding for a new transcription factor (ftf1) of Fusarium oxysporum is only expressed during infection of common bean. Fungal Genetics and Biology. 2007;44(8):864-876.
4. Butler EJ. Fungi and diseases of plants. Wallingford (UK): CABI Publishing; 1918. p. 233-270.
5. De RK, Chaudhary RG, Naimuddin. Comparative efficacy of biocontrol agents and fungicides for controlling chickpea wilt caused by Fusarium oxysporum f. sp. ciceri. The Indian Journal of Agricultural Sciences. 1996;66(6):370-373.
6. Dubey SC, Suresh M, Birendra S. Evaluation of Trichoderma species against Fusarium oxysporum f. sp. ciceri for integrated management of chickpea wilt. Journal of Biological Control. 2007;40(1):118-127.
7. Haware MP, Nene YL, Natrajan M. Survival of Fusarium oxysporum f. sp. ciceri in soil in absence of chickpea. In: Proceedings of the National Seminar on Management of Soil Borne Diseases of Crop Plants; 1986 Jan 8-10; Coimbatore, Tamil Nadu, India. Tamil Nadu Agricultural University; 1986.
8. Kumar A, Tripathi SC. Evaluation of the leaf juice of some higher plants for their toxicity against soil borne pathogens. Plant and Soil. 1991;132(2):297-301.
9. Nene YL, Thapliyal PN. Fungicides in plant disease control. 3rd ed. New Delhi: Oxford and IBH Publishing Company; 1993. p. 531-532.
10. Nene YL, Haware MP, Reddy MV. Chickpea diseases: resistance-screening techniques. Information Bulletin No. 10. Patancheru (AP), India: International Crops Research Institute for the Semi-Arid Tropics; 1981.
11. Padwick GW. Plant protection and food crops of India (Plant pests and diseases of rice, wheat, sorghum and gram). Empire Journal of Experimental Agriculture. 1948;60:55-64.
12. Poonia TC, Pithia MS. Pre-and post-emergence herbicides for weed management in chickpea. Indian Journal of Weed Science. 2013;45(3):223-225.
13. Ravichandran S, Hegde YR. Evaluation of fungicides against Fusarium oxysporum f. sp. ciceri causing chickpea wilt. Chemical Science Review and Letters. 2015;4(16):1042-1046.
14. Rojo FG, Reynoso MM, Sofia MF, Chulze N, Torres AM. Biological control by Trichoderma species of Fusarium solani using peanut brown root rot under field conditions. Crop Protection. 2007;26(4):549-555.
15. Singh KB, Dahiya BS. Breeding of wilt resistance in chickpea. In: Symposium on Wilt Problem and Breeding for Wilt Resistance in Bengal Gram; 1973 Sep; New Delhi, India. Indian Agricultural Research Institute; 1973. p. 13-14.
16. Singh RS, Singh D, Singh HV. Fungal antagonistics on chickpea wilt caused by Fusarium oxysporum f. sp. ciceri. Plant Disease Research. 1977;12:103-107.
17. Sonawane SS, Pawar NB. Studies on biological management of chickpea wilt. Journal of Maharashtra Agricultural Universities. 2001;26(2):215-216.