Dissecting Fusarium wilt resistance in chickpea (Cicer arietinum L.) through genotypic screening under artificial conditions

Introduction

Chickpea (Cicer arietinum L.) is a self-pollinated, diploid (2n = 16) cool-season grain legume of significant global importance (Varshney et al., 2013) [13] . It is broadly classified into two major groups, desi and kabuli, which differ in morphological, physiological and genetic attributes, including seed characteristics, stress tolerance and adaptation patterns (Jimenez-Diaz et al., 2015) [7] . Chickpea is valued for its high nutritional content, serving as an affordable source of protein, dietary fiber and essential micronutrients, particularly in developing countries (Gaur et al., 2012) [3] . It also contributes to sustainable agriculture through symbiotic nitrogen fixation, improving soil fertility under rainfed and marginal conditions (Narayan et al., 2024) [8] . Chickpea is widely cultivated across diverse agro-ecological regions (Hattiholi et al., 2024) [4] , while India is a leading producer globally, with about 10.24 million hectares area under cultivation (73.78% of the world area) and production of 11.68 million tonnes (73.45%) (DAC&FW, 2024-25) [2] . Chickpea, despite being a valuable source of nutrition and income, is highly vulnerable to a range of biotic and abiotic stresses that considerably hamper its productivity. Important yield limiting factors include fungal diseases such as Ascochyta blight and Fusarium wilt, pod borer infestation and unfavorable environmental conditions like drought and cold stress (Sankar et al., 2024) [10] . Fusarium oxysporum f. sp. ciceris is the causal agent of Fusarium wilt in chickpea, which poses a major constraint to its productivity (Haware et al., 1978) [6] . Under congenial environmental conditions, the disease can lead to yield losses approaching 100 percent (Pande et al., 2011) [9] . Disease incidence has been reported to reach up to 61 percent during the vegetative stage and around 43 percent at the flowering stage, indicating its severe impact across crop growth phases. Yield losses are more pronounced when infection occurs at early growth stages, with early wilt accounting for approximately 77-94% reduction, compared to 24-65% under late wilt conditions. Furthermore, infection during the reproductive phase, particularly from flowering to pod development, can result in drastic reductions in seed weight, with losses reported as high as 90%, underscoring the destructive nature of the disease. Several high yielding varieties such as Annigeri, JG 62, that are preferred by farmers are susceptible to Fusarium wilt. Hence, the cultivation of wilt-resistant varieties of chickpea is the best way to overcome yield and production losses. In view of this, present investigation was carried out to isolate the potential wilt resistant chickpea genotypes to aid in disease resistance broadly in chickpea.

Materials and Methods Isolation and multiplication of pathogen

Diseased portion was washed under running tap water. Small infected tissue pieces were cut using a sterile blade, surface sterilized with 1% sodium hypochlorite and rinsed thrice in sterile distilled water. The tissues were aseptically placed on Potato Dextrose Agar (PDA) plates and incubated at 27±1 °C. Fungal growth was monitored and pure cultures were obtained using the hyphal tip method and maintained on PDA slants. Pathogenicity of the isolated fungus was confirmed by inoculating susceptible host plants and observing characteristic disease symptom development. For long-term maintenance, the cultures were periodically sub-cultured at monthly intervals and preserved on PDA slants at 4 °C (Sharma et al. 2010) [11] .

Screening of Chickpea Genotypes under Controlled Conditions

Forty two chickpea genotypes were screened for response to Fusarium oxysporum f. sp. ciceri infestation under controlled greenhouse conditions. The experiment was laid out in a completely randomized design (CRD) with two replications during rabi 2024-25. Two standard checks, WR 315 (R) and JG 62 (HS), were included to validate the screening procedure. The experiment was conducted using pots filled with sterilized soil to ensure uniform growth conditions. The pathogen inoculum was prepared on sorghum meal medium in conical flasks. The substrate was sterilized on two consecutive days to eliminate contaminants. A mycelial disc of 5 mm from a 7 day old actively growing culture of F. oxysporum f. sp. ciceri was aseptically transferred into each flask and incubated for 15 days at suitable temperature conditions, with intermittent shaking to ensure uniform colonization. The fully colonized substrate was mixed thoroughly and incorporated into the soil in each pot at an inoculum level of 8% (w/w). Five seeds were sown per pot. Prior to sowing, seeds of each genotype were surface sterilized with 1% sodium hypochlorite for 1 minute, followed by repeated washing with sterile distilled water for 2 times. Observations on wilt incidence were recorded periodically based on plant mortality. The percent disease incidence (PDI) was calculated using the following formula: Based on the percent disease incidence, genotypes were categorized into different reaction groups following the scale proposed by Haware and Nene (1982) [5] with slight modifications (Table . 1). The study revealed significant variability among the test genotypes for wilt disease reaction indicating the presence of diverse reaction levels. There were genotypes viz DCPV 21-23, DCPV 21-85, DCPV 21-131 and WR 315 which showed resistant reaction, suggesting their potential as reliable sources of resistance. Moderately resistant genotypes exhibiting superior yield performance may be recommended for cultivation, while also serving as valuable genetic sources in breeding programmes focused on the development of durable and broad-spectrum disease resistant chickpea cultivars. The genotypes categorized as moderately susceptible to highly susceptible, including popular cultivars like Annigeri and JG 62, exhibited the vulnerability under wilt conditions (Fig. 1) and cultivation of such entries under wilt prone fields should be curtailed. The clear differentiation of genotypes under controlled inoculation confirms the effectiveness of the screening method. Similar findings were obtained by Ashwini et al. (2024) [1] , where 25 genotypes were screened for wilt resistance under greenhouse conditions. Takalakki et al. (2025) [11] identified 10 germplasm lines and 7 advanced breeding lines resistant to wilt under artificial sick plot condition. The present findings were aligned with results of Yadav et al. (2023) [13] . The identified resistant and moderately resistant genotypes can be utilized in breeding programmes for developing fusarium wilt resistant chickpea varieties.

Conclusion

Fusarium wilt remains a major constraint in chickpea production due to its destructive nature and widespread occurrence. In the present study, genotypes exhibiting resistant and moderately resistant reactions to Fusarium oxysporum f. sp. ciceri were isolated, which can be a valuable sources of resistance. These genotypes can be utilized both for direct deployment in wilt prone areas based on yield performance and as donors in breeding programmes aimed at developing resistant cultivars. Upon generating the molecular insights on resistance and tagging the markers for resistance aids in enhancing the efficiency of crop improvement through marker assisted selection. In addition, regular screening of diverse germplasm along with continuous monitoring of pathogen variability will be crucial for achieving durable and broad based resistance in chickpea.

References

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