Integration of Trichoderma, Pseudomonas and fungicides for the control of collar rot disease of chickpea (Cicer arietinum L.)

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International Journal of Agriculture, Environment and Biotechnology Citation: IJAEB: 10(1): 12531, February 201 DOI: 10.595/2230-32X.201.00005.5 201 New Delhi Publishers. All rights reserved Integration of richoderma, Pseudomonas and fungicides for the control of collar rot disease of chickpea (Cicer arietinum L.) Shyam Singh* 1, V.K. Nirmalkar 2, R.K.S. iwari 3, Anjeet Jangre 4 and Pankaj Kumar 5 1 Department of Plant Pathology, S.K. College of Agriculture and Research Station, Indira Gandhi Krishi Vishwavidyalaya, Kawardha- Kabirdham, Chhattisgarh 491995,India 2 Department of Plant Pathology, K.V. K., Indira Gandhi Krishi Vishwavidyalaya, Bilaspur, Chhattisgarh 495001,CG,India 3 Department of Plant Pathology,.C.B. College of Agriculture and Research Station, Indira Gandhi Krishi Vishwavidyalaya, Bilaspur - 495001,CG,India 4 Department of Agronomy, S.K. College of Agriculture and Research Station, Indira Gandhi Krishi Vishwavidyalaya, Kawardha- Kabirdham, Chhattisgarh 491995,India 5 Department of Plant Pathology, College of Agriculture, Indira Gandhi Krishi Vishwavidyalaya, Raipur 492012,CG,India *Corresponding author e-mail: shyamsingh.skcars@gmail.com Paper No.: 560 Received: 1-02-2016 Accepted: 0-02-201 Abstract An experiment was conducted on integration of richoderma, Pseudomonas and fungicides for the control of Collar rot disease of Chickpea during 20134 and 20145. Results indicated that the most effective treatment was richoderma harzianum @ q/ha 1 (Soil) + Hexaconazole @ 3ml/kg 1 seed with minimum mortality (4.30 and 2.25%) which was at par with treatment Pseudomonas fluorescens @ q/ha 1 (Soil) + Hexaconazole @ 3ml/kg 1 seed (5.0 and 2.59%) and richoderma harzianum @ q/ha 1 (Soil)+ubeconazole @ 3ml/kg 1 seed (6.15 and 4.09%) whereas maximum mortality 15.0 and 12.35%) was recorded in control plot. Maximum no. of pods per plant (41.30 and 49.5) was recorded in treatment = richoderma harzianum @ q/ha 1 (Soil) + Hexaconazole @ 3ml/kg 1 seed which was at par with = Pseudomonas fluorescens @ q/ ha 1 (Soil) + Hexaconazole @ 3ml/kg 1 (3. and 45.95) and significantly superior over rest of the treatment. In case of grain yield highest grain yield was increased in treatment = richoderma harzianum @ q/ha 1 (Soil) + Hexaconazole @ 3ml/kg 1 seed (44.5%) followed by = Pseudomonas fluorescens @ q/ha 1 (Soil) + Hexaconazole @ 3ml/kg 1 (43.61%) and 10 = Pseudomonas fluorescens @ q/ha 1 (Soil) + ubeconazole @ 3ml/kg 1 seed (29.63%). Highlights PLAN PAHOLOGY richoderma harzianum @ q/ha 1 (Soil)+Hexaconazole @ 3ml/kg 1 seed and Pseudomonas fluorescens @ q/ ha 1 (Soil)+Hexaconazole @ 3ml/kg 1 was found most effective treatment for the control of collar rot of chickpea under field condition. As compare to alone application richoderma harzianum or Pseudomonas fluorescens or Hexaconazole integrated application of richoderma harzianum or Pseudomonas fluorescens and Hexaconazole showing minimum mortality and maximum plan growth and yield of chickpea under field condition. Keywords: Chickpea, Collar rot, Integration management, Pseudomonas fluorescens, richoderma harzianum Chickpea (Cicer arietinum L.) is the most important pulse crop in India. India is the largest producer of chickpea with about 63% of the total area under chickpea production lying in India. Gram is the most dominant pulse having a share of around 40% in the total production followed by pigeonpea at 15 to 20% and urdbeen and mungbeen around 0%. Area and production and productivity of chickpea in India have been increased during the past decade. In India, chickpea occupies an area of mha and its

Singh et al. production is.1 mt with an average productivity of 5 kg ha 1 during 20145 (Anonymous 2016) but the production was decreased as compared to 20134 (9.5 million tons) due to many biotic and abiotic constraints. Nearly 12 pathogens have been reported so far that infect chickpea (Cicer arietinum L.) in different parts of the world but only a few of them have the potential to devastate the crop. Some diseases are persistent problems in chickpea production in wide geographical areas, notably, ascochyta blight, fusarium wilt, dry root rot, botrytis gray mold, collar rot, black root rot, phytophthora root rot, and pythium root and seed rot, while others are sporadic in occurrence or endemic in distribution (Nene et al. 2012). Among them soil borne diseases such as fusarium wilt (Fusarium oxysporum f. sp. ciceris), dry root rot (Rhizoctonia bataticola), collar rot (Sclerotium rolfsii) and black root rot (Fusarium solani) are the major limiting factor in chickpea production (Ghosh et al. 2013). Among the above soil borne pathogen Sclerotium rolfsii is the major problematic fungus in Chhattisgarh. Now days it is becoming a serious problem in central and peninsular India due to high soil moisture, the presence of under decomposed organic matter on the soil surface, low soil ph and high temperature (25 to 30 C) which are favor the disease development (Al-Askar et al. 2013). It has an extensive host range; at least 500 species in 100 families are susceptible (Nagamma and Nagaraja 2015). he most common hosts are the legumes, crucifers, and cucurbits. S. rolfsii commonly occurs in the tropics, subtropics, and other warm temperate regions including India. Sclerotium rolfsii Sacc., is a major cause for 55 95% mortality of chickpea seedlings (Gurha and Dubey 192). his may be due to the prolific growth, extensive host range of the pathogen and having the ability to produce large number of sclerotia that may persist in the soil for several years (Sennoi et al. 2013). Several management strategies such as physical, chemical, biological and growing of resistant varieties were used for the control of collar rot disease caused by Sclerotium rolfsii Sacc.. Still there is no resistant or tolerant varieties were identified against this pathogen in India. In case of chemical control large numbers of broad range or spectrum fungicides were tried by the several workers but there is no one fungicides was much effective under field condition. Besides of this fungicides are damaging the biotic and abiotic environments which are problematic for human beings. he control of collar rot of chickpea (Sclerotium rolfsii) is not possible by one method of plant disease control. It can be manage by the integration of all available control measures. herefore, keeping in view the above facts an experiments was conducted on integration of richoderma, Pseudomonas and fungicides for the control of Collar rot disease of Chickpea. Materials and methods An experiment was conducted on integration of richoderma, Pseudomonas and fungicides for the control of Collar rot disease of Chickpea at S.K. College of Agriculture and Research Station Farm, Kawardha (Kabirdham), C.G. during 20134 and 20145. Experiment layout under Randomized Block Design (RBD) with ten treatment and three replications. he plot size of experiment was 4x 3 m. Chickpea variety JG6 is widely grown in Chhattisgarh was selected for this experiment. Culture of richoderma harzianum and Pseudomonas fluorescens were collected from Department of Plant Pathology, CB college of Agriculture and Research Station (IGKV), Bulaspur, C.G. richoderma harzianum culture was maintained on PDA media and Pseudomonas fluorescens culture was maintained in King s B medium and stored at 5 C. he mass culture of richoderma harzianum and Pseudomonas fluorescens was prepared on alk powder as per the standard procedure adopted by State Biological Control Laboratory (SBCL), CB College of Agriculture and Research Station, Chorbhatti, Bilaspur (C.G.). Seeds were treated with ubeconazole @ 3ml, Hexaconazole @ 3ml, richoderma harzianum @ 10g and Pseudomonas fluorescens @ 10g as per the treatments details given in able 1. In case of soil application. harzianum and P. fluorescens antagonists were inoculated in the sterilized FYM at 20 days before sowing. his inoculated FYM kept under shade to allow grow of both bio-control agents. After the colonization enriched FYM applied in furrow before sowing of the crop. hen the furrows were covered with soil. Seeds of variety JG6 were sown with row to row distance 30 cm and plant to plant 10 cm. 126

Integration of richoderma, Pseudomonas and fungicides for the control of collar rot disease... Observation on mortality was recorded at weekly interval in which total and rotted plants were recorded from each experimental plot. Plant height, number of pod and grain yield were also recorded from the all experimental plot. he per cent diseased incidence was calculated with following formula: reatment code 1 2 3 4 5 Per cent disease incidence = Number of plants infected otal number of plants examined 100 able 1: reatment Details Control reatment details Seed treatment with ubeconazole @ 3ml Seed treatment with Hexaconazole @ 3ml Seed treatment with Pseudomonas fluorescens @ 10g Seed treatment with richoderma harzianum @ q ha +10g 6 Soil application of P. fluorescens @ q ha + Seed treatment with P. fluorescens 10g kg seed Soil application of. harzianum @ q ha + Seed treatment with Hexaconazole @ 3ml Soil application of P. fluorescens @ q ha + Seed treatment with Hexaconazole @ 3ml 9 Soil application of. harzianum @ q ha + Seed treatment with ubeconazole @ 3ml 10 Soil application of P. fluorescens @ q ha + Seed treatment with ubeconazole @ 3ml Results and discussion Experimental data indicated that the minimum mortality recorded was 4.30 and 2.25% in plot treated with richoderma harzianum @ q ha (Soil) + Hexaconazole @ 3ml followed by plot treated with Pseudomonas fluorescens @ q ha (Soil) + Hexaconazole @ 3ml (5.0 and 2.59 %) during 20134 and 20145, respectively whereas, untreated plot showed maximum mortality of 15.0 and 12.35% during 20134 and 20145, respectively. Among the all treatments treatment mean highest reduction in plant mortality was recorded in plots treated with richoderma harzianum @ q ha (Soil) + Hexaconazole @ 3ml (6.66%) followed by plot treated with Pseudomonas fluorescens @ q ha (Soil) + Hexaconazole @ 3ml (0.10%) (able 2). Maximum mean plant height was observed in treatment richoderma harzianum @ q ha + 10 g (Soil + Seed) (49.03 cm) followed by Pseudomonas fluorescens @ q ha +10g (Soil + Seed) (4.50 cm) and richoderma harzianum @ q ha (Soil) + Hexaconazole @ 3ml (4.3 cm). here is no significantly difference recorded among the all treatment (able 3). Data pertaining to no. of pods per plant have been presented in able 3 reveal that the maximum no. of pods per plant (41.30 and 49.5) was recorded in treatment = richoderma harzianum @ q ha (Soil) + Hexaconazole @ 3ml which was at par with = Pseudomonas fluorescens @ q ha (Soil) + Hexaconazole @ 3ml/ (3. and 45.95) and significantly superior over rest of the treatment. While, minimum number of pods per plant (25.40 and 36.0) was observed in untreated control. In case of grain yield maximum grain yield (14.269 and 19.095 q ha ) was observed in = richoderma harzianum @ q ha (Soil) + Hexaconazole @ 3ml followed by = Pseudomonas fluorescens @ q ha (Soil) + Hexaconazole @ 3ml kg (14.12 and 1.94 q ha ) while minimum grain yield (14.135 q ha ) was obtained in untreated control. Maximum grain yield was increased in treatment = richoderma harzianum @ q ha (Soil) + Hexaconazole @ 3ml (44.5%) followed by = Pseudomonas fluorescens @ q ha (Soil) + Hexaconazole @ 3ml kg (43.61%) and = Pseudomonas fluorescens @ q 10 ha (Soil) + ubeconazole @ 3ml (29.63%) (able 4). Result indicated that the combine application fungicides as seed treatment and bio-control agents as soil treatment was found effective in controlling of collar rot disease of chickpea. his might be due to initial effect of fungicides and long time affectivity of bio-control agents. Sugha et al. (1993) also reported that conidial coating of the antagonistic richoderma harzianum and. viride on seeds significantly reduced seedling mortality (4-65%) infected by Sclerotium rolfsii compared with the untreated controls. Khan and Javaid (2015) tested egula 12

Singh et al. able 2: Efficacy of richoderma, Pseudomonas and fungicides against collar rot disease of chickpea Plant mortality (%) 20134 20145 Mean Reduction in plant mortality (%) 15.0 (23.2) 12.35 (20.53) 14.03 (21.90) -.0 (16.30) 5.4 (13.34) 6. (14.2) 51.1.41 (16.66) 5.2 (13.6).0 (15.1) 49.64 12.35 (20.4) 9.60 (1.9) 10.9 (19.23) 21. = richoderma harzianum @ q ha (Soil) + 10g kg seed 9.54 (1.3).22 (15.4).3 (16.65) 40.2 = Pseudomonas fluorescens @ q ha (Soil) + 10g kg seed 10.2 (19.0).95 (1.33) 9.9 (1.20) 29.54 = richoderma harzianum @ q ha (Soil) + Hexaconazole @ 3ml kg seed 4.30 (11.) 2.25 (.53) 3.2 (10.20) 6.66 = Pseudomonas fluorescens @ q ha (Soil) + Hexaconazole @ 3ml/kg seed 5.0 (13.) 2.59 (.32) 4.20 (11.10) 0.10 = richoderma harzianum @ q ha (Soil) + ubeconazole 9 @ 3ml kg seed 6.15 (14.30) 4.09 (11.35) 5.12 (12.3) 63.51 = Pseudomonas fluorescens @ q ha (Soil) + ubeconazole 10 @ 3ml kg seed.32 (15.66) 5.06 (12.9) 6.19 (6.19) 55. CV (%) 11.50 13.46 12.4 SEm± 0.1 0.95 0.3 CD at 5% 1.6 2.4 2.1 reatments = Control 1 = ubeconazole @ 3ml kg seed 2 = Hexaconazole @ 3ml kg seed 3 = Pseudomonas fluorescens @ 10g kg seed 4 5 6 richoderma harzianum @ q ha (Soil) + Hexaconazole @ 3ml/kg seed Control Pseudomonas fluorescens @ q ha (Soil) + Hexaconazole @ 3ml kg seed Overall view of experiment 12

Integration of richoderma, Pseudomonas and fungicides for the control of collar rot disease... able 3: Efficacy of richoderma, Pseudomonas and fungicides against collar rot disease of chickpea reatments Plant height (cm) No. of pods per plant 20134 20145 Mean 20134 20145 Mean = Control 40.25 45.60 42.93 25.40 36.0 31.05 1 = ubeconazole @ 3ml 41.30 46.5 44.03 32.00 43.50 3.5 2 3 4 5 = Hexaconazole @ 3ml 42.00 4.65 44.3 35.50 43.5 39.63 = Pseudomonas fluorescens @ 10g 44.45 49.50 46.9 29.30 40.90 35.10 = richoderma harzianum @ q ha (Soil) + 10g 46.25 51.0 49.03 30.50 41.40 35.95 = Pseudomonas fluorescens @ q ha (Soil) + 10g kg 6 seed = richoderma harzianum @ q ha (Soil) + Hexaconazole @ 3ml = Pseudomonas fluorescens @ q ha (Soil)+Hexaconazole @ 3ml/kg seed 46.00 51.00 4.50 26.24 3.0 31.9 45.50 49.95 4.3 41.30 49.5 45.53 45.00 49.15 4.0 3.0 45.95 42.33 = richoderma harzianum @ q ha (Soil) + ubeconazole 9 @ 3ml = Pseudomonas fluorescens @ q ha (Soil) + 10 ubeconazole @ 3ml 44.65 4.00 46.33 36.20 44.65 40.43 43.60 4.60 45.60 36.0 45.05 40.93 CV (%) 12.52 14.52 14.50 12.46 SEm± 2.40 2.5 3.23 2.9 CD at 5% NS NS.40.52 able 4: Efficacy of richoderma, Pseudomonas and fungicides against collar rot disease and grain yield of chickpea reatments Grain Yield (q/ha) 20134 20145 Mean Increased yield over control (%) = Control.96 14.135 11.516-1 = ubeconazole @ 3ml 11.019 16.065 13.542 1.59 2 3 4 5 6 = Hexaconazole @ 3ml 11.11 16.245 13.61 1.0 = Pseudomonas fluorescens @ 10g 10.19 15.33 12.59 10.0 = richoderma harzianum @ q ha (Soil) +10g 10.91 16.043 13.50 1.29 = Pseudomonas fluorescens @ q ha (Soil) +10g 10.244 15.635 13.303 15.52 = richoderma harzianum @ q ha (Soil)+Hexaconazole @ 3ml = Pseudomonas fluorescens @ q ha (Soil)+Hexaconazole @ 3ml/ kg seed = richoderma harzianum @ q ha (Soil)+ubeconazole @ 3ml 9 = Pseudomonas fluorescens @ q ha (Soil)+ubeconazole @ 3ml 10 14.269 19.095 16.62 44.5 14.12 1.94 16.53 43.61 11.3 16.965 14.402 25.06 12.431 1.425 14.92 29.63 CV (%) 10.2 9. 10.0 - SEm± 0.9 0.6 0. - CD at 5% 2.33 2.24 2.29-129

Singh et al. (tebuconazole), hiophanate Methyl, Ridomil Gold (metalaxyl + mancozeb) and Mancozeb fungicides at 50, 100, 150, 200, 250 ppm concentrations. Fungicides hiophanate methyl, egula and Mancozeb were found effective against Sclerotium rolfsii. Kumar et al. (200) evaluated two fungicides (carboxin and thiram) and two bio-control agents (Pseudomonas fluorescens and richoderma harzianum) as seed treatment in different combinations against Sclerotium rolfsii, the causal organism of collar rot of chickpea (Cicer arietinum). Seed treated with. harzianum (4g/kg seed) + carboxin (0.5g/kg seed) provided maximum protection to the crop by giving maximum seedling emergence (495.0/20 m 2 ), final plant stand (40.4/20m 2 ) and grain yield (1.2q/ha). Nagamma and Nagaraja (2015) revealed that the maximum inhibition of mycelial growth (1.6%) was noticed in. harzianum (Bacteriology lab isolate) which was followed by. viride (Microbiology lab) (63.33%). Least inhibition was observed in. harzianum GKVK isolate (31.6%). Sab et al. (2014) tested eight bioagents tested against S. rolfsii. richoderma harzianum-55 IIHR recorded maximum inhibition of 0% followed by. harzianum NBAII with 63% and least mycelial inhibition was observed in Pseudomonas fluorescens and Bacillus subtilis. Veena and Reddy (2016) revealed that the seed treatment with Copper oxychloride + soil application of potential fungal (richoderma isolate-) and bacterial bio-control agent (CREB6) was found to be superior as it recorded the highest germination percentage (100 %), highest initial (10.00) and final population of chickpea (9.66), least PDI (16.00 %), maximum plant height (25.61 cm), root length (13.40 cm) and maximum shoot (0.49 g) and root dry weights (0.11 g). he population dynamics of both antagonists and pathogen were estimated at two different intervals initially at DAS and then on 45 DAS in pot culture experiment. With the increase in antagonists population, the population dynamics of pathogen were significantly reduced from DAS to 45 DAS over control in all the treatments with the maximum reduction in treatment. Singh et al. (2013) revealed that the richoderma harzianum NBRI055 significantly suppress the seedling blight of sunflower caused by Rhizoctonia solani and induce defence mechanism in host. Singh et al. (2014) revealed that the seed and 130 furrow application of the formulation of richoderma viride 2% W.P. formulation significantly reduced the wilt disease of tomato and damping-off of chilli. he yields of tomato and chilli were also significantly enhanced. Further, the formulation did not have any phyto-toxic effect either on tomato or chilli plants at all the doses levels tested for field bio-efficacy. he. viride formulation also did not have any adverse effect on the beneficial rhizospheric microbes, like Arbuscular Mycorrhizae (Glomus spp.) in tomato and chilli rhizosphere at all dosages as confirmed with microscopic observations. Based on the above findings, the. viride 2% W.P. formulation is found safe and effective for using as an efficient and ecologically-safe alternative to chemical fungicides for the management of wilt of tomato and dampingoff of chilli as well as for obtaining higher yields. Conclusion he main objective of the present study was to manage the collar rot disease of chickpea by integration of fungicide and richoderma harzianum or Pseudomonas fluorescens. he results of the present study concluded that the collar rot disease of Chickpea can be minimizing by the integration of richoderma harzianum @ q ha (Soil application) or Pseudomonas fluorescens q ha (Soil application) followed by the application of Hexaconazole @ 3ml as seed treatment under field conditions. References Al-Askar, A.A., Rashad, Y.M. and Absulkhair, W.M. 2013. Antagonistic activity on an endemic isolate of Streptomyces tendae RDS 16 against phytopathogenic fungi. J Mycobiol Resist., 6: 509-516. Anonymous. 2016. Report of commodity Profile for Pulses April 2016. 10. Ghosh, R., Sharma, M., elangre, R. and Pande, S. 2013. Occurrence and Distribution of Chickpea Diseases in Central and Southern Parts of India. American Journal of Plant Sciences 4: 940-944. Gowdra, N. and Nagaraja, A. 2015. Efficacy of biocontrol agents against Sclerotium rolfsii causing collar rot disease of chickpea, under in vitro conditions. Internat J Plant Protec., (2): 222-22. Gurha, S.N. and Dubey, R.S. 192. Occurrence of possible sources of resistance in chickpea (Cicer arietinum L.) against Sclerotium rolfsii Sacc. Madras Agriculture Journal 0: 63-64. Harman, G.E. 2000. Myths and dogmas of biocontrol: changes in perceptions derived from research on richoderma harzianum 22. Plant Disease 4: 3-393.

Integration of richoderma, Pseudomonas and fungicides for the control of collar rot disease... Khan, I.H. and Javaid, A. 2015. Chemical control of collar rot disease of chickpea. Pakistan Journal of Phytopathology 2(1): 61-6. Kumar, R., Mishra, P., Singh, G. and Yadav, R.S. 200. Integration of bioagent and fungicides for management of collar rot of chickpea. Journal of Biological Control 22(2): 4-49. Neeraja, B., Kumar, V.M., Prasadji, J.K. and Kumar, P.A. 2014. Effect of agrochemicals on richoderma harzianum (H4 isolate) and its biocontrol potential against chickpea collar rot caused by Sclerotium rolfsii. International Journal of Food, Agriculture and Veterinary Sciences 4(3): 1333. Sab, J., Nagaraja, A. and Nagamma, G. 2014. Efficacy of biopesticides against Sclerotium rolfsii Sacc. causing collar rot of chickpea (Cicer arietinuml.). he Bioscan: International Quarterly Journal of Life Science 9(1): 335-339. Sennoi, R., Jogloy, S., Saksirirat, W., Kesmala,. and Patanothai, A. 2013. Genotypic variation of resistance to southern stem rot of Jerusalem artichoke caused by Sclerotium rolfsii. Euphytica 190: 415-424. Singh, B.N., Singh, A., Singh, B.R. and Singh, H.B. 2013. richoderma harzianum elicits induced resistance in sunflower challenged by Rhizoctonia solani. Journal of Applied Microbiology 116(3): 654 666. Singh, H.B., Singh, A., Sarma, B.K. and Upadhyay, D.N. 2014. richoderma viride 2% W.P. (Strain No. BHU- 2953) Formulation Suppresses omato wilt Caused by Fusarium oxysporum f. sp. lycopersici and Chilli Dampingoff Caused by Pythium aphanidermatum effectively under Different Agroclimatic Conditions. International Journal of Agriculture, Environment & Biotechnology (2): 313-320. Sugha, S.K., Sharma, B.K. and yagi, P.D. 1993. Factors affecting development of collar rot of gram (Cicer arietinum) caused by Sclerotium rolfsii. Indian J Agril Sci., 63(6): 32-35. Veena, G.A. and Reddy, N.P.E. 2016. Integrated disease management of dry root rot of chickpea. International Journal of Applied Biology and Pharmaceutical echnology (2): 45-54. 131