Management of seed borne mycoflora of proso millet by using fungicides and bioagents

Introduction

Proso millet (Panicum miliaceum L.) classified within the tribe Paniceae under family Poaceae (Graminae) with subfamily Panicoideae. It is a fourploid species having four sets of chromosome number totaling 36. (Rajesekaran et al., 2023) [1] The crop is extensively cultivated across several countries, including India, China, Japan, Arabia, Egypt, and regions of Western Europe. In India, it is predominantly cultivated in Andhra Pradesh, Bihar, eastern Uttar Pradesh, Karnataka, Madhya Pradesh, Maharashtra, and Tamil Nadu. Small millets are primarily grown in semi-arid regions like India as rainfed crops. Despite its beneficial qualities small millet production makes up less than 2% of global cereal production. The genetic potential of small millets is significantly constrained by both biotic and abiotic stresses, causing significant yield and economic loss. Among biotic stress factors, diseases cause by fungal pathogen such as blast, leaf spot, leaf blight, rusts and smuts are major challenges in small millet growing areas. (Ramesh et al., 2024) [2] . Pathogen free seeds serve as a prerequisite for successful crop establishment, fostering uniform germination, healthy seedling emergence, and ultimately greater agricultural output. The seed borne mycoflora of proso millet comprises multiple fungi, including Fusarium moniliforme, Bipolaris panici-miliacei, Curvularia lunata, Aspergillus niger, Aspergillus flavus, Aspergillus ochraceus, and Cladosporium cladosporioides. These pathogens are critical contributors to poor germination and yield losses. Treating seeds with fungicides, biocontrol agents, and other appropriate methods is an effective strategy for managing seed borne fungal pathogens, improving seed health, enhancing crop performance and ultimately increasing yield. Fungicides remain one of the most widely used approaches for controlling fungal diseases, while biological control agents offer environmentally friendly alternatives by suppressing the growth of pathogenic fungi. (Chaudhari et al., 2017) [3] . Therefore the present investigation was conducted to study the management of seed borne mycoflora of proso millet using fungicides and bioagents.

Materials and Methods

The investigation was conducted under in vitro laboratory condition during 2025-26 at the Department of Plant Pathology, College of Agriculture, Nagpur, Maharashtra, India. Seeds of proso millet varieties TNAU-202 and TNAU-145 were used for the detection of seed borne mycoflora. Seed borne mycoflora associated with proso millet seeds were detected using method prescribed by (ISTA, 1976) [4] namely, standard blotter paper method and standard agar plate method. Seven different fungicides were evaluated under in vitro against test pathogen by adopting poison food technique. The treatments evaluated include carbendazim 25% + mancozeb 50% WS, penflufen 13.28% + trifloxystrobin 13.28% FS, carboxin 37.5% + thiram 37.5% DS, prochloraz 34.8% + propiconazole 7.8% EC, tebuconazole 5.4% FS, fluxapyroxad 333 g/l FS, metalaxyl 8% + mancozeb 64% WP and an untreated control. Potato dextrose agar (PDA) medium was prepared and sterilized in autoclave. Requisite quantity of each of the fungicide (as per concentration) was added in sterilized, melted PDA separately to obtain desired concentration. Flask containing poisoned medium was shaken well to have even and uniform distribution of fungicides. About 20 ml of melted poisoned PDA was poured in each sterilized petri plate and allow to solidify. The petri plates were inoculated separately by test fungus. Five mm disc of one week old fungus culture was cut with sterilized cork borer, lifted and transferred aseptically in the centre of petri plate containing the medium poisoned with test fungicide. The control plates were kept the culture disc grown in same condition on PDA without fungicides. Treated plates were incubated at room temperature (26 ± 2 °C) for a period of seven days. Colony diameter was recorded in mm as per Vincent's formula based on the average colony diameter. The data was subjected to statistical analysis wherever necessary. Four bioagents were evaluated under in vitro condition against test pathogen by adopting dual culture method. The treatments include Trichoderma asperellum, Trichoderma harzianum, Pseudomonas fluorescens, Bacillus subtilis, and an untreated control. PDA was autoclaved and poured into 90 mm petri plate, where it solidified. Then, on one end of the plate a 5 mm disc of test species was set and on the other end, an antagonistic disc and striking bacterial culture on both sides where a 5 mm disc of test pathogen is placed in centre. Along with the control, the plates were incubated at room temperature (26 ± 2 °C) for seven days. The data was subjected to statistical analysis wherever necessary. The data recorded were analyzed statistically by Completely Randomized Design for all the experiment in vitro and critical difference (CD) were calculated at 1 per cent probability.

Result and Discussion

Proso millet seed samples were examined using method prescribed by (ISTA, 1976) [4] namely standard blotter paper method and standard agar plate method. Based on morphological and microscopic observation the fungi observed viz., Fusarium moniliforme, Bipolaris panicimiliacei, Curvularia lunata, Aspergillus niger, Aspergillus flavus, Aspergillus ochraceus and Cladosporium cladosporioides.

Poisoned food technique

The efficacy of seven fungicides was evaluated in vitro at their respective concentration against seed borne mycoflora of proso millet. The data is presented and tabulated in Table 1 [5] and Tammineni et al. (2025) [6] .

2. Bipolaris panici-miliacei: Among the seven fungicides, carboxin 37.5% + thiram 37.5% DS and prochloraz 34.8% + propiconazole 7.8% EC achieved complete mycelial growth inhibition of 100% followed by fluxapyroxad 333 g/l FS recorded 81.53%, tebuconazole 5.4% FS recorded 80.76%, metalaxyl 8% + mancozeb 64% WP recorded 76.92% and carbendazim 25% + mancozeb 50% WS recorded 76.15%. Penflufen 13.28% + trifloxystrobin 13.28% FS was the least effective with mycelial growth inhibition of 68.21%. The highest growth of Bipolaris panici-miliacei was recorded in the control. The findings of present investigation in agreement with Gupta et al. (2025) [7] .

3. Curvularia lunata: Among the seven fungicides, carboxin 37.5% + thiram 37.5% DS and prochloraz 34. [8] . [8] .

Aspergillus ochraceus:

Dual culture method

The efficacy of four biocontrol agents viz., Trichoderma asperellum, Trichoderma harzianum, Pseudomonas fluorescens and Bacillus subtilis, was evaluated against seed borne mycoflora of proso millet by dual culture method. Results obtained are tabulated in Table 2.

1. Fusarium moniliforme: Maximum mycelial growth inhibition of Fusarium moniliforme was observed in Trichoderma asperellum (75.71%) followed by Trichoderma harzianum (68.92%) and Bacillus subtilis (61.42%). Minimum mycelial growth inhibition was observed in Pseudomonas fluorescens (36.61%). The findings of present investigation are in agreement with Meghavathu et al. (2025) [5] and Tammineni et al. (2025) [6] .

Bipolaris panici-miliacei:

Maximum mycelial growth inhibition of Bipolaris panici-miliacei was observed in Trichoderma asperellum (80.74%) followed by Trichoderma harzianum (66.17%) and Bacillus subtilis (57.21%). Minimum mycelial growth inhibition was observed in Pseudomonas fluorescens (50.74%). The findings of present investigation are in agreement with Gupta et al. (2025) [7] .

3. Curvularia lunata: Maximum mycelial growth inhibition of Curvularia lunata was observed in Trichoderma asperellum (73.07%) followed by Bacillus subtilis (69.23%), Trichoderma harzianum (65.38%). Minimum mycelial growth inhibition was observed in Pseudomonas fluorescens (56.15%). The finding of present investigation in agreement with Tekade et al. (2017) [9] .

Aspergillus niger: Maximum mycelial growth inhibition of

Aspergillus niger was observed in Trichoderma harzianum (74.14%) followed by Trichoderma asperellum (57.95%), Bacillus subtilis (44.31%). Minimum mycelial growth inhibition was observed in Pseudomonas fluorescens (03.52%). The present investigation in agreement with Sutar et al. (2024) [8] 5. Aspergillus ochraceus: Maximum mycelial growth inhibition of Aspergillus ochraceus was observed in Bacillus subtilis (79.26%) followed by Trichoderma asperellum (65.05%), and Trichoderma harzianum (60.79%). Minimum mycelial growth inhibition was observed in Pseudomonas fluorescens (25.28%). The findings of present investigation are in agreement with Sutar et al. (2024) [8] .

Conclusion

References

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8. Siren R, Zope AV, Bulle SR, Totawar MV. Comparative evaluation of fungicides against seed mycoflora of wheat. J. Pl. Dis. Sci. 2025;20(2):211-219. How to cite this article: Kale AA, Bramhankar S, Pillai TS, Gathe A, Peshattiwar P, Shinde GV. Management of seed borne mycoflora of proso millet by using fungicides and bioagents. International Journal of Advanced Biochemistry Research. 2026;SP-10(9):2591-2594. doi: https://www.doi.org/10.33545/26 174693.2026.v10.i9Saf.10223