INTRODUCTION:

Chickpea (Cicer arietinum L.) is an important pulse crop of family Leguminaceae. It is used as a big source of protein in the human diet. Chickpea is one of the best legumes for human consumption

It is an important crop of Indian sub-continent that usually contributes more than 66%in terms of global production, while In India, chickpea is ranked first in terms of production and consumption in the world. About 65% of global area with 68% of global production of chickpea is contributed by India (Amarender and Devraj, 2010).

Low yield of chickpea is attributed to its susceptibility to several fungal, bacterial and viral diseases. Fusarium wilt caused by Fusarium oxysporum. f. sp. ciceri (Padwick) Matuo and K. Sato, is the most important soil-borne disease of chickpea throughout the world and particularly in the Indian Subcontinent, the Mediterranean Basin and California (Nene and Reddy, 1987).

Trichoderma spp. generally grows in its natural habit on plant root surface and therefore it controls root diseases in particular (Monte, 2001; Faruk et al., 2002; Kamlesh and Gujar, 2002). The species of Trichoderma have been evaluated against the wilt pathogen and have exhibited greater potential in managing chickpea wilt under field condition (Podder et al., 2004).

The disease can appear at any stage of plant growth, symptoms in a highly susceptible cultivar can develop any time between 25 days after sowing till as late as podding stage (Nene 1985). The disease has assumed great importance.Annual yield losses in chickpea were estimated to be 4.8 million tones worldwide due to biotic stresses, including infectious plant diseases (Ryan, 1997).

In India it is 10–15%, which in years of severe epidemics may rise to 60–70% (Jalali and Chand, 1992). Mycoparasites (Papavizas, 1985) and many bacteria (Weller, 1988) have shown promising results in managing phytopathogenic fungi. P. fluorescens has revolutionised the field of biological control of soil-borne plant pathogenic fungi (Burr et al., 1998). That bacterium produces phenazin (Toohey et al., 1965; Gurusiddaiah et al., 1986), pyrolnintrin (Burkhead and Geoghegan, 1994), phloroglucinol (Howell and Stipanovic, 1980) and siderophores (Sakthivel et al., 1986), which may be involved in the suppression of the wilt fungus (Fridlender et al., 1993; Gamliel and Katan, 1993). Leeman et al. (1995) reported satisfactory control of Fusarium wilt of radish by treating the seed with P. fluorescens. In addition, P. fluorescens produces auxins, gibberellins etc. (Glick, 1995) and solubilises phosphorus in the soil (Dube and Yeole, 1997), which helps plant growth. Among mycoparasites,

The genus Trichoderma includes the most widely used biocontrol agent of soil-borne, seed-borne and other diseases (Chet et al., 1979; Chet and Baker, 1981). Trichoderma harzianum and T. virens are active rhizosphere colonisers (Tronsmo andHarman, 1992) that produce antibiotics such asgliotoxin, viridin, and some cell wall degrading enzymes (Larito et al., 1976; Bello et al., 1997) and also certain biologically active heat-stable metabolites such as ethyl acetate (Claydown et al., 1987). These substances may be involved in disease suppression or plant growth promotion. Trichoderma harzianum is one efficient biocontrol agent that is successfully used to suppress Fusarium wilt (Khan et al., 2004; Dubey et al., 2007). Similarly, amending soil with plant extracts significantly reduces Fusarium wilt in the field (Chand and Singh, 2005).To study the effect of fungicides on disease incidence.To study the effect of fungicides on test fungus.To study the effect of bio control agents on disease incidence

MATERIALS AND METHODS:

Cleaning and sterilization of glass wares:

The glasswares (petri dishes, pipettes, conical flasks, test-tube) used in the experiment will be thoroughly washed with detergent powder and air dried. The Petri dishes and pipettes will be wrapped in clean paper and sterilized in hot air oven at temperature of 150⁰C for two hours.

Isolation and identification of pathologen:

Fusarium oxysporium f.sp.ciceriused in the the experiment will isolated in pure culture from the root of infected chickpea plant showing characteristics symptoms of wilt disease .the infected roots of chickpea will be wash three time sterilized with 0.1% Hgcl₂for 1-2 second. Previously before culturing infected parts will view under microscope for ascertaining examination of conidia. By applying Hgcl₂ tissue get surface sterilized so as to minimize the contamination. Already sterilized melted PDA will transferred into Petri-dishes and then small pieces of chick pea infected roots will keep on semi-solidify media inside petriplates .This whole process will be done inside the laminar flow under highly asceptic condition. These Petri-plates will be incubated under room temperature.

Viability and population assessment test of the product:

Commercial formulations of Trichoderma viride and Pseudomonas fluorescence will tested for the viability and population assessment test before using in the experiment by the following procedure.1g of product will weight and make upto10 ml with sterilized distilled water and was shaken well (1:10).1ml of this suspension will take and transferred to 9 ml of sterilized water in a test tube(1:100) serial dilution will made similarly transferring 1ml of the suspension to the subsequent tubes to get 1:1000000 dilution 1ml.of the 1000000 suspension will transferred to sterile pertiplate .15 ml of the melt and cool PDA will poured in Petri plates for assessment Trichoderma viride and Pseudomonas fluorescence. The plates will incubated at room temperature .after 48 hrs average no of colonies per plate will be calculated.

No of colonies

cfu in/g product=-------------------------x Dilution factor

Amount place

Characters of Trichoderma viride :

Conidiophores hyline much branched, not verticillatePhialides single or in groupConidia (phialospores) hyaline, 1-celled, ovoid, borne in small terminal clustersUsually easily recognized by its rapid growth and green patches or cushions of conidia.Saprophytic in soil or wood very common some species reported as parasites on other fungi

Bacterial biocontrol agent:-

Pseudomonas fluorescence is another bio-control agent and will used in experiment for controlling wilt of chick pea (gram) caused by Fusarium oxysporum f.sp.ciceri.

Incorporation of bacterial antagonists Pseudomonas fluorescence into medium:

Effect of P. fluorescence on growth of test fungus Fusarium oxysporum f.sp.ciceri. will studied using different concentration of bacterial suspension prepared from 48 h old culture of bacterial isolate grown on kings’B medium. Bacterial suspension will used at (0.1ml) and (0.3ml) concentrations. Transfer different concentrations bacterial suspension in replicated petriplates and poured the medium into sterilized petri plate as the rate of 15 ml of mediumper dish and allow to solidify.

Inoculation of test organisms:-

A 5 mm disc of Fusarium oxysporum f.sp.ciceri will cut with a sterile cork borer and transfer asceptically to the center of medium contained in a petridish. Control will maintain with the mycelia disc of the Fusarium oxysporum f.sp.ciceri. on PDA medium containing bacterial suspension. All the treatments will be replicated 3 times. First trial will be conducted during 2015-16 second trial will conducted from 2016-17.

Inoculation of fungus in PD broth:-

All the treatments will be keep according as in radial growth method. The only difference between the two experiments will be here the same fungus was inoculated in PD broth carried in conical flasks. After the inoculation of the fungus in all the conical flasks will inoculated under room temp. all the treatments replicate 5 time.

Disease intensity (%) was calculated by using the following formula:

Sum of all disease rating

Disease intensity (%)=---------------------------X 100

Total no rating x Max disease grade

(IRRI, 1996)

RESULTS ANDDISCUSSION:

The results of study entitled “Efficacy of fungi toxicants and bio-control agents against gram wilt caused by Fusariumoxysporum ciceri “under “In vitro” conditions were conducted in the Department of Plant Protection Allahabad Agricultural Institute Deemed University, Allahabad are included in tables and figures are presented

Effect of treatments:

During both the years of experimentation Trichoderma viride in 2:1 ratio resulted in significantly reduced the growth of Fusarium oxysporumf. Sp. ciceri followed by Trichoderma vidide in 1:1 ratio. The growth of Fusarium oxysporumf. Sp. Ciceri was the highest in the inoculated test fungus treatments. During both the years of experimentation Pseudomonas fluorescence in the 0.3 ml resulted in significantly reduced the growth of Fusarium oxysporum f. Sp. Ciceri followed by Pseudomonas fluorescence 0.1 ml. The growth of Fusarium oxysporumf. Sp. Ciceri was the highest in the inoculated test funges treatments. During both the years of experimentation, Neem leaf extract 6% resulted insignificantly reduced the growth of Fusarium oxysporum f. Sp. ciceri followed by Neem leaf extract 3%. The growth of Fusarium oxysporum f. Sp. ciceri was the highest in the inoculated fest fungus treatments. The order of the treatments T₂<T₁<T₀During both the year of experimentation Neem bark extract 6% resulted in significantly reduced the growth of Fusarium oxysporum f. Sp. ciceri followed by Neem bark extract 3%. The growth of Fusarium oxysporum f. Sp. ciceri was the highest in the inoculated test fungus treatments.

Interaction effect:

During 2015-16, 2016-2017 treatment T₂significantly reduced the redial growth of Fusarium oxysporumf. Sp. ciceri as compared with control at all the successive stages of the radial growth expert T₂D₁ and ToD₁ were ate per with each other. Similar observation was also recorded in case of T₁ during both the years of experimentation. During 2015-2016, and 2016-2017 treatment T2 significantly inhibited the radial growth of Fusarium oxysporum f. Sp. ciceri as compared with control at all the successive stages of the radial growth. Similar results was observed in T₁ also. During 2015-2016, and 2016-2017 Neem leaf extract 6% T₂ significantly inhibited the radial growth of Fusarium oxysporum f. Sp. ciceri as compared with control at all the stages of the radial growth. Neem leaf extract 3% (T₁) also significantly reduced the radial growth of Fusarium oxysporum f. Sp. Ciceri at all the stages of radial growth as compared with control during both the years of experimentation. Neem leaf extract (6%) also reduced the radial growth of Fusarium oxysporum f.sp. ciceri significantly as compared with neem leaf extract (3%) at D₅, D₆, D₇ and D₈ stages during 2015-2016 and at D₃, D₄, D₅, D₆, D₇ and D₈ during 2016-17. During 2015-2016, and 2016-2017 treatment T₂significantly inhibited the radial growth of Fusarium oxysporum f. Sp. ciceri as compared with control at all the successive stages of the radial growth. Similar results was observed in T₁ also.

Effect of Bavistin, Thiram on growth of Fusarium oxysporum f. Sp. ciceri by radial growth method

Effect of treatments:

During both the years of experimentation Bavistin 100 ppm resulted in significantly reduced the growth of Fusarium oxysporum f. Sp. ciceri followed by Bavistion 10 ppm. The growth of Fusarium oxysporum f. Sp. ciceri was the highest in the inoculated test fungus treatments. The order of the treatments T2<T1<T0 During both the years of experimentation Thiram 100 ppm resulted in significantly reduced the growth of Fusarium oxysporum f. Sp. ciceri followed by Thiram 10 ppm. The growth of Fusarium oxysporum f. Sp. ciceri was the highest in the inoculated test fungus treatments. The order of the treatments T2<T1<T0

Interaction effect:

During 2015-2016-2016-2017 Bavistin 100 ppm (T₂) significantly inhibited the growth of Fusarium oxysporum f. Sp. ciceri as compared with control at all the stages of the radial growth. Bavistin 10 ppm (T₁) also reduced significantly the radial of Fusarium oxysporum f. Sp. ciceri at all the stages of radial growth as compared with control during both the years of experimentation. Bavistin 100 ppm also reduced the radial growth of Fusarium oxysporum f. Sp. ciceri significantly as compared with Bavistin 10 ppm at all the stages of radial growth during the years of experimentation. During 2015-2016, and 2016-2017 .Thiram 100 ppm (T₂) significantly inhibited the growth of Fusarium oxysporum f. Sp. ciceri as compare with control at all the stages of the radial growth. Thiram 10 ppm (T₁) also reduced significantly the radial growth of Fusarium oxysporum f. Sp. ciceri at all the stages of radial growth as compared with control during both the years of experimentation. Thiram 100 ppm also reduced the radial growth of Fusarium oxysporum f. Sp.ciceri significantly as compared with Thiram 10 ppm at all the stages of radial growth during both the year of experimentation.

Comparing the growth of Fusarium oxysporum f. Sp. ciceri with Trichoderma viride in PD broth.

During both the years of experimentation data showed that after 12 days of inoculation of fungus in PD broth, the mycelium of Fusarium oxysporum f. Sp. ciceri was completely overgrown by Trichoderma viride in both treatments T₁ (Fusarium oxysporum f. Sp. ciceri+Trichoderma viride ration of 1:1) and T₂(Fusarium oxysporum f. Sp. ciceri+Trichoderma viride ration of 2:1), where as in T₀Fusarium oxysporum f. Sp. ciceri showed its normal growthThe present ïn vitro” study indicated that the growth of Fusarium oxysporum f. Sp. ciceri ws completely inhibited by Trichoderma viride in liquid medium.

Effect of Pseudomonas fluorescence, Neem leaf extract, Neem bark extract, Bavistin, Thiramon growth of Fusarium oxysporum f. Sp. ciceri at 12 DAI by mycelia weight method.

During both the years of experimentation data showed that after 12 days of inoculation of fungus in PD broth, Pseudomonas fluorescence 0.3ml resulted in significantly reduced the growth of Fusarium oxysporum f. Sp. ciceri followed by Pseudomonas fluorescence 0.1 m. The growth of Fusarium oxysporum f. Sp. ciceri was the highest in the inoculated test fungus treatments. The order of treatments was T2<T1<T0

During both the years of experimentation, data showed that after 12 days of inoculation of fungus in PD broth, Neem leaf extract 6% resulted in significantly reduced the growth of Fusarium oxysporum f. Sp. ciceri followed by Neem leaf extract 3%. The growth of Fusarium oxysporum f. Sp. ciceri was the highest in the inoculated test fungus treatments. The order of treatment was T2<T1<T0During both the years of experimentation, data showed that after 12 days of inoculation of fungus in PD broth, Neem bark extract 6% resulted in significantly reduced the growth of Fusarium oxysporum f. Sp. ciceri followed by Neem bark extract 3%. The growth of Fusarium oxysporum f. Sp. ciceri was the highest in the inoculated test fungus treatments. The order of treatments was T2<T1<T0During both the years of experimentation data showed that after 12 days of inoculation of fungus in PD broth, Bavistin 100 ppm resulted in significantly reduced the growth of Fusarium oxysporum f. Sp. ciceri followed by Bavistin 10 ppm. The growth of Fusarium oxysporum f. Sp. ciceri was the highest in the inoculated test fungus treatments. The order of treatments was T2<T1<T0During with the years of experimentation, data observed showed that after 12 days of inoculation of fungus in PD broth, Thriam 100 ppm resulted in significantly reduced the growth of Fusarium oxysporum f. Sp. ciceri followed by Thiram 10 pp,. The growth of Fusarium oxysporum f. Sp. ciceri was the highest in the inoculated test fungus treatments.The order of treatment was T2<T1<T0

There were significant different in the shoot length (cm) of chickpea due to different treatments at 30, 60 and 90 DAS during both the years of experimentation.At 30 DAS the maximum shoot length was recorded in the T₃ (Fusarium oxysporum f. Sp. ciceri+Pseudomonas flurescence with 19.78 cm and 15.50 cm of chickpea during 2015-2016, and 2016-2017 and it was followed by T₂Fusarium oxysporum f. Sp. ciceri+Trichoderma viride with 17.30 cm and 12.50 cm of chickpea during 2015-2016, and 2016-2017 and the minimum shoot length was recorded in the T1 (Fusarium oxysporum f. Sp. ciceri) with 5.00 cm and 5.10 cm of chickpea during 2015-2016, and 2016-2017 respectively.At 60 DAS the maximum shoot length was recorded in the T₃ (Fusarium oxysporum f. Sp. ciceri+Pseudomonas fluorescence ) 41.20 cm and 38.10 cm of chickpea during 2015-2016, and 2016-2017 and it was followed by T2 (Fusarium oxysporum f. Sp. ciceri+Trichoderma viride) 36.8 cm and 35.00 cm of chickpea during 2015-2016, and 2016-2017 respectively and the minimum shoot length was recorded in the T₁ (Fusarium oxysporum f. Sp. ciceri) 13.50 cm 13.33 cm of chickpea during 2015-2016, and 2016-2017 respectively.The order of Treatments during both the year wasT₃>T₂>T₄>T₅>T₇>T₆>T₈>T1at 90 DAS the maximum shoot length was recorded in the T₃ (Fusarium oxysporum f. Sp. ciceri+ Pseudomonas fluorescene ) 52.30 cm and 52.00 cm of chickpea during 2015-2016, and 2016-2017 respectively and it was followed by T2 (Fusarium oxysporum f. Sp. ciceri+Trichoderma viride ) 47.87 cm and 47.70 cm of chickpea during 2015-16 and 2016-17 respectively and the minimum shoot length was recorded in the T₁ (Fusarium oxysporum f. Sp. ciceri) 15.50 cm and 15.38 cm of chickpea during 2015-2016, and 2016-2017 respectively.The order of Treatment during both the years was T₃>T₂>T₄>T₅>T₇>T₆>T₈>T₁at 30 DAS, the maximum root length was recorded inthe treatment T₃ (Fusarium oxysporum f. Sp. ciceri+Pseudomonas fluorescence having 7.35 cm and 7.30 cm of chickpea 2015-2016, and 2016-2017 respectively and it was followed by T₂ (Fusarium oxysporum f. Sp. ciceri+Trichoderma viride) having 6.80 and 6.33 cm of chickpea during 2015-2016, and 2016-2017 respectively and the minimum root length was recorded in the R1 Fusarium oxysporum f. Sp. ciceriinoculated ) having 4.60 cm and 4.27 cm of chickpea during 2015-2016, and 2016-2017 respectively.The order of treatment during both the years was T₃>T₂>T₄>T₅>T₇>T₆>T₈>T₁

At 60 DAS the maximum root length was recorded in the treatment T3 (Fusarium oxysporum f. Sp. ciceri+Pseudomonas fluorescence having 15.06 cm and 15.13 cm of chickpea during 2015-2016, and 2016-2017 respectively and it was followed by T2 (Fusarium oxysporum f. Sp. ciceri+Trichoderma viride) having 12.73 cm and 12.83 cm of chickpea during 2015-2016, and 2016-2017 respectively and the minimum root length was recorded in the T1 (Fusarium oxysporum f. Sp. ciceri inoculated) having 5.27 cm and 5.27 cm of chickpea 2015-2016, and 2016-2017 respectively. The order of treatment during both the years was T3>T2>T4>T5>T7>T6>T8>T1 At 90 DAS the maximum root length was recorded in the T3 (Fusarium oxysporum f. Sp. ciceri+Pseudomonas fluorescence having 17.53 cm and 17.83 cm of chickpea during 2015-2016, and 2016-2017 respectively and it was followed by T2 (Fusarium oxysporum f. Sp. ciceri+Trichoderma viride) having 16.17 cm and 16.47 cm of chickpea during 2015-2016, and 2016-2017 respectively and the minimum root length was recorded in the T1 (Fusarium oxysporum f. Sp. ciceri inoculated ) having 5.43 cm and 5.47 cm of chickpea during during 2015-2016, and 2016-2017 respectively. The order of treatment during 2015-16 was T3>T2>T4>T5>T7>T6>T8>T1. The order of treatment during 2016-17 T3>T2>T4>T5>T7> T6>T8>T1. There were significant differences in the shoot weight (g) of chickpea due to different treatment at 30, 60 and 90 DAS during both the years of experimentation. At 30 DAS, the maximum shoot weight in (g) was recorded in the T3 (Fusarium oxysporum f. Sp. ciceri+Pseudomonas fluorescence) having 21.40 g and 21.70 g of chickpea during 2015-2016, and 2016-2017 respectivelyand it followed by T2 (Fusarium oxysporum f. Sp. ciceri+Trichoderma viride) having 18.60 g and 19.00 g of chickpea during 2015-2016, and 2016-2017 respectively and the minimum shoot weight was recorded in the T1 (Fusarium oxysporum f. Sp. ciceri) havng 3.20 g and 3.63 g of chickpea during 2015-2016, and 2016-2017 respectivelyThe order of Treatments during 2015-16 was T3>T2>T4>T5>T7>T6>T8>T1. The order of Treatment during 2016-17 T3>T2>T4>T5>T7>T6>T8>T1

At 60 DAS the maximum shoot weight was recorded in the T3 (Fusarium oxysporum f. Sp. ciceri+Pseudomonas fluorescence ) having 39.60 g and 39.30 g of chickpea during 2015-2016, and 2016-2017 respectively and it was followed by T2 (Fusarium oxysporum f. Sp. ciceri+Trichoderma viride) having 33.5 g and 32.6 g of chickpea during 2015-2016, and 2016-2017 respectively and the minimum shoot weight was recorded in the T1 Fusarium oxysporum f. Sp. ciceri) having 6.60 g nad 7.10 g of chickpea during 2015-2016, and 2016-2017 respectively The order of treatment during 2015-16 was T3>T2>T4>T5>T7>T6>T8>T1. The order of treatment during 2016-17 was T3>T2>T4>T5>T7>T6>T8>T1 At 90 DAS the maximum shoot weight was recorded in the T3 (Fusarium oxysporum f. Sp. ciceri+Pseudomonas fluorescence ) having 49.70g and 49.80 g of chickpea during 2015-2016, and 2016-2017 respectively and it was followed by T2 (Fusarium oxysporum f. Sp. ciceri+Trichoderma viride) having 47.06 g and 47.50 g of chickpea during 2015-2016, and 2016-2017 respectively and the minimum shoot weight was recorded in the T1 (Fusarium oxysporum f. Sp. ciceri) having 7.60 g and 8.30 g of chickpea during 2015-2016, and 2016-2017 respectively. The order of Treatment during both the years was T3>T2>T4>T5>T7>T6>T8>T1 At 30 DAS the maximum root weight was recorded in the T3 Fusarium oxysporum f. Sp. ciceri+Pseudomonas fluorescence) having 20.60 g na d21.30 go of chickpea during 2015-2016, and 2016-2017 respectively and it was followed by T2 (Fusarium oxysporum f. Sp. ciceri+Trichoderma viride)Having 15.30 g and 15.40 g of chickpea during 2015-2016, and 2016-2017 respectively and the minimum root weight was recorded in the T1 (Fusarium oxysporum f. Sp. ciceri) having 1.90 g and 2.20 g of chickpea during 2015-2016, and 2016-2017 respectively/The order ot treatments during both the years was T3>T2>T4>T5>T7>T6>T8>T1

At 60 DAS the maximum root weight was recorded in The T3 (Fusarium oxysporum f. Sp. ciceri+Pseudomonas flurescence) having 23.30 g and 23.2 g of chickpea during 2015-2016, and 2016-2017 respectively and it was followed by T2 (Fusarium oxysporum f. Sp. ciceri+Trichoderma viride) having 21.40 g and 21.70 g of chickpea during 2015-2016, and 2016-2017 respectively and the minimum root weight was recorded in the T1 (Fusarium oxysporum f. Sp. ciceri) having 4.8 g and 4.5 g of chickpea during 2015-2016, and 2016-2017 respectively The order of Treatments during both the years was T3>T2>T4>T5>T7>T6>T8>T1

At 90 DAS the maximum root weight was recorded in the T3 (Fusarium oxysporum f. Sp. ciceri+Pseudomonas fluorescence ) having 35.10 g and 35.17 g of chickpea during 2015-2016, and 2016-2017 respectively and it was followed by T2 (Fusarium oxysporum f. Sp. ciceri+Trichoderma viride ) having 32.37 g and 33.17 g of chickpea during 2015-2016, and 2016-2017 respectively and the minimum root weight ws recorded in the T1 (Fusarium oxysporum f. Sp. ciceri) having 6.60 g and 6.67 g of chickpea during 2015-2016, and 2016-2017 respectively The order of Treatments during both the years was T3>T2>T4>T5>T7>T6>T8>T1

SUMMARY AND CONCLUSION:

Chickpea is the most important winter (Rabi) pulse crop, cultivated in India. Chickpea in general also play a unique role in restoring the soil fertility by denitrogen fixing through symbiosis of root nodules bacteria of Rhizbium species. This crop is badly affected by many diseases, out of them one os wilt disease caused by Fusarium oxysporum f. sp. ciceri which is most severe among all of them. Keeping in view the severity of damage and its control measure, the present problem was undertaken in the Department of Plant Pathology, Sam Higginbottom University Agriculture Science and Technology, Allahabad, in which Management of chick pea wilt caused by Fusarium oxysporum f. sp.ciceri was studied both “In vitro” and In vivo” conditions. The experiment “In vitro” was carried out in the laboratory Departmentof Plant Pathology Sam Higginbottom University of Agriculture Science and Technology. The fungus causing wilt disease was isolated from the chickpea roots showing characteristics symptoms of Fusarium infection. The two bio control agents used are viz. Trichoderma viride and Pseudomonas flurescence, fungitoxicants viz. Neem leaf extract, Neem bark extract, Bavistin and Thiram. These tasted in both solid and broth medium.The test fungus isolated was inoculated on PDA and PD- broth. The commercial formulation of Trichoderma viride and Pseudomonas fluorescence were tested for their viability and population assessment before using in the experiment by Serial dilution techniques Pure culture of Trichoderma viride and Pseudomonas flurescence was maintained by periodic sub culturing in PDA and PD-broth, King’s B medium and King’s B broth by zig-zag streaking after every 15 day respectively.In radial growth method three replicates in the PDA medium was maintained for each treatment including control.

The radial growth of fungal colony (in mm) was recorded every 24 hrs. Till eight days from the date of inoculation. In mycelia weight method five replicated was maintained for each treatments including control. The mycelia weight (in g) of test fungus was recorded at 12 days after inoculation in broth medium.The fungi-toxicants used in radial growth method, three replicates in the PDA medium was maintained for each treatments including control.In mycelia weight method five replicates in the PD broth was maintained for each treatment including control and mycelia weight of test fungus was recorded at 12 DAI in broth medium.

Soil inoculation with Fusarium oxysporum f. sp. ciceri @ 100g/plot of soil. Seed dressed with bio-control agents @ 4g/ Kg of seed and fungitoxicants viz. Neem leaf and Neem bark powder @ 4g/Kg of seed, Bavistin and Thiram@3 g/Kg of seed. Observation of wilt incidence was recorded at 30, 60 and 90 DAS. Root length, Shoot length, Root weight and Shoot weight were also recorded at 30, 60 and 90 DAS. The wilt incidence was recorded minimum with Bavistin, Thiram and Trichoderma viride.The Shoot weight, Root weight, Shoot length and Root length was maximum when treated with Pseudomonas fluorescence (T₃), Trichoderma viride (T₂).

FUTURE TIME OF ACTION/ RECOMMENDATION:

In view of present exploratory studies conducted in “In vitro” and “In vivo” at Department of Plant Protection Allahabad Agricultural Institute Deemed University, Allahabad, it can be conducted that Bavistin, Thiram, Trochoderma viride and Pseudomonas fluorescence in hibit the growth of Fusarium oxysporum f. sp. ciceri. But the chemicals provide good short term protection, and the biological fungus provides long term root protection As a consequence, yields frequently are increased over use of the chemical alone chemicals are very toxic for controlling soil-borne disease like Fusarium oxysporum f. sp. ciceri. Which are very hazardous to our flora and fauna. The main problems caused from pesticides are:

1) It disturbs our ecological equilibrium

2) It results in increase of human diseases

3) It decreases number of defenders as natural enemies

4) It has temporary effect

For avoiding these factors, bio-control agents should be use which have eco-friendly non- poisonous behaviour and do not adversely effect the crop. But it still needs more investigation to be conducted in this regards for proper recommendation

REFERENCES:

1. Amarender R, Devraj M (2010)., Growth and instability in chickpea production in India.www.krisat.org Accessed on 15 February 2011.

2. Bendre NJ Barhate BG (1998)., A Souvenir on disease management in chickpea. M.P.K.V. Rahuri during 10th Dec. 1998.

3. Ankita Shukla and S.K.Dwivedi (2012).,Bioefficacy of plant extracts against fusarium species causing wilt in pulses Vol. 2 Issue 1, Jan.2012, pp. 136-144

4. Bendre NJ Barhate BG (1998)., A Souvenir on disease management in chickpea. M.P.K.V. Rahuri during 10th Dec. 1998.

5. Burr A., A. Ortuno and T. Armero, (1998)., Phosphate solubilizing effect of Aspergillus niger and Pseudomonas.Microbiologia Espanola 30, 113.

6. Burkhead K. and M.J. Geoghegan, (1994)., Antibiotics. In: Soil-borne Plant Pathogens. (K. Burkhead ed.) Macmillon, New York, NY, USA, 368 pp.

7. Bello D.K., H.D. Wells and C.R. Morkhan, (1997)., In vitroantagonism of Trichoderma species against six fungalplant pathogens. Phytopathology 72, 579.

8. Cho S Muehlbauer FJ (2004)., Genetic effect of differentially regulated fungal response genes on resistance to necrotrophic fungal patho-gens in chickpea (Cicer arietinum L.). Physiol. Mol. Plant Pathol. 64: 57–66.Cook RJ (1985)., Biological control of plant pathogens: theory to application. Pathopathology 12:75-80.

9. Chet I Y. Hadar. Y. Elad., J. Katan and Y. Henis, (1979)., Biological control of soil-borne pathogens by Trichodermaharzianum. In: Soil Borne Plant Pathogens (B. Schippers ed.), Academic press, London, UK 585

10. Chand H, Singh S (2005)., Control of chickpea wilt (Fusarium oxysporum f sp ciceri) using bioagents and plant extracts. Indian J. Agric. Sci. 75: 115-116.

11. Claydown K.L., O.H. Emerson and R.J. Sauthwell, (1987)., The isolation of a toxic substance from the culture filtrate of Trichoderma. Phytopathology 36, 1068

12. .Dubey SS, Suresh M, Singh B (2007)., Evaluation of Trichoderma species against Fusarium oxysporum f. sp. ciceris for integrated management of chickpea wilt. Biol. Control 40(1):118-127.

13. Dube H.C. and R.D. Yeole, (1997)., Increased plant growths and yield through seed bacterization. Indian Phytopathology 50 (3), 316–319.

14. Dubey SC, Suresh M, Singh B (2007)., Evaluation of Trichoderma species against Fusarium oxysporum f. sp. ciceris for integrated management of chickpea wilt. Biol. Control 40: 118-127.

15. Faruk MI, Rahman, ML, Bari MA (2002)., Management of seedling disease of cabbage through Trichoderma harzianum amendment in seedbed. Bangl. J. Plant Pathol. 18(1-2):49-53.

16. Monte E (2001). Understanding Trichoderma: between biotechnology and microbial ecology. Int. Microb. 4:1-4.

17. Fridlender M., J. Inbar and I. Chet, (1993)., Biological controlof soilborne pathogens by a β-13 glucanase producing Pseudomonas cepacia. Soil Biology and Biochemistry25, 1211–1221.

18. Grewal JS (1969)., Important fungal disease of Cicer arietinum in India. Pulse Improvement Project Seminar Report held at Karaj Agricultural College, University of Tehran & USDA, January 7-9, 1969. pp. 35-40.

19. Gurusiddaiah S., D.M. Weller., A. Sarkar and R.J. Cook,(1986)., Characterization of an antibiotic produced by a strain of P. fluorescens inhibitory to Gaemannomyces graminis var. tritici and Pythium spp. Antimicrobialagents and Chemotherapy 29, 488–495.

20. Gamliel A and J. Katan, (1993)., Suppression of major and minor pathogens by fluorescent pseudomonads in solarized and nonsolarized soil. Phytopathology 83(1), 68-75

21. Glick B.R, (1995)., The enhancement of plant growth by free living bacteria. Canadian Journal of Microbiology 41, 109–117.

22. Haware MP, Nene YL, Mathur SB (1986)., Seed borne diseases of chickpea. Technical Bulletin 1. Danish Government Institute of seed Technology for developing countries. Copenh. (1):1-32.

23. Harman GE, Charles RH, Ada V, Chet I, Matteo L (2004)., Trichoderma -opportunistic, avirulent plant symbionts. Nat. Rev. Microbiol. (2):43-56. Haware

24. Harman GE (2006), Overview of mechanism and uses of Trichoderma spp. Phytopathol. (96):190-194.

25. Howell C.R and R.D. Stipanovic, (1980)., Suppression of Pythium utlimum-induced damping-off of cotton seedlingsby Pseudomonas fluorescens and its antibiotic pyoluteorin. Phytopathology 70, 712–715.

26. Hanan Ibrahim Mudawi1, , Mohamed Osman Idris(2014).,The efficacy of Trichoderma spp. and Bacillus isolates in the control of chickpea wilt pathogens Agriculture, Forestry and Fisheries 2014; 3(5): 346-351

27. Jayalakshmi S.K, Raju S, Usha Rani S, Benagi V.I and Sreeramulu K(2009).,Trichoderma harzianum L1 as a potential source for lytic enzymes and elicitor of defense responses in chickpea (Cicer arietinum L.) against wilt disease caused by Fusarium oxysporum f. sp. ciceri.

28. Jalali, B.L. and H. Chand. (1992)., Chickpea wilt. In: Plant Diseases of International Importance. Vol. I. Diseases of Cereals and Pulses, (Eds.): Singh, U.S., A.N. Mukhopadhayay, J. Kumar and H.S. Chaube. Prentice Hall, Englewood Cliffs, NJ. pp. 429-444.

29. Kamlesh M, Gujar RS (2002)., Evaluation of different fungal antagonistic, plant extracts and oil cakes against Rhizoctonia solani causing stem rot of chilli seedlings. Ann. Plant Prot. Sci. 10(2):319-322.

30. Khan MR, Khan SM, Mohiddin FA (2004)., Biological control of Fusarium wilt of chickpea through seed treatment with the commercial formu-lation of Trichoderma harzianum and/ or Pseudomonas fluorescens. Phytopathol. Mediterr. 43: 20-25

31. Leeman M., J.A. vanPelt., M.K. Hendrickz., R.J.Scheffe., P.A.H.M. Bakker and B. Schippers, (1995)., Biocontrol of Fusarium wilt of radish in commercial green house trials by seed treatment with Pseudomonas fluorescens WCS 374. Phytopathology 85, 1301–1305.

32. Larito P., J. Webster and N. Lomas, (1976). Trichoderma viride produce gliotoxin and viridin. Transactions of British Mycological Society 47, 535

33. Md. Motaher Hossain1, Nilufar Hossain1, Farjana Sultana, Shah Mohammad Naimul Islam, Md. Shaikul Islam1 and Md. Khurshed Alam Bhuiyan(2013)., Integrated management of Fusarium wilt of chickpea (Cicer arietinum L.) caused by Fusarium oxysporum f. sp. ciceris with microbial antagonist, botanical extract and fungicide Vol. 12(29), pp. 4699 4706,

34. Muneeb andrabi, amrish vaid, vijay kumar razdan (2010)., Evaluation of different measures to control wilt causing pathogens in chickpeaVol. 51, No. 1

35. Mujeebur R. Khan, Shahana M. Khan And Fayaz A. Mohiddin (2004)., Biological control of Fusarium wilt of chickpea through seed treatment with the commercial formulation of Trichoderma harzianum and/or Pseudomonas fluorescens Phytopathol. Mediterr. (2004) 43, 20–25

36. Muhammad Nasir Subhani1, Shahbaz Talib Sahi, Liaqat Ali, Safdar Hussain1, Javaid Iqbal1 and Nisar Hussain1 (2013)., Management of Chickpea wilt caused by Fusarium oxysporium f. sp. ciceris through antagonistic microorganisms Canadian Journal of Plant Protection. Volume 1, Number 1, Pages 1-6 ©2013 Canadian Science and Technology Press Inc

37. Nene YL, Reddy MV (1987)., Chickpea Diseases and their Control. In: Saxena M.

38. Nene Y.L. (1985)., Opportunities for research on disease of pulse crops. Indian Phytopathol. 38: 1–10.

39. Podder RK, Singh DV, Dubey SC (2004)., Integrated application of Trichoderma harzianum mutants and carbendazim to manage chickpea wilt (Fusarium oxysporum f. sp, ciceri). Ind. J. Agric. Sci. 74:346-348.

40. Papavizas G.C,(1985).,Trichoderma and Gliocladium: biology,ecology and potential for control. Annual Review ofPhytopathology 23, 23–54

41. R.k. Poddar*, d.v. Singh and s.c. Dubey (2014)., Management of chickpea wilt through combination of fungicides and bioagents Indian Phytopath. 57 (1) : 39-43

42. Ryan, J.G. (1997)., A global perspective on pigeon pea and chickpea sustainable production systems-present status andfuture potential. In: Recent Advances in Pulses Research. (Eds.): Asthana A.P. and M. Ali. Indian Society of Pulses Research and Development, Kanpur, India. pp. 1-31.

43. Shabir-U-Rehman1, W. A. Dar, S. A. Ganie, Javid A. Bhat, Gh. Hassan Mir, Rubina Lawrence1, Sumati Narayan and Pardeep Kumar Singh(2013).,Comparative efficacy of Trichoderma viride and Trichoderma harzianum against Fusarium oxysporum f sp. ciceris causing wilt of chickpea vol.7(50),pp. 5731-5736

44. Sakthivel N., E. Sivamani., N. Unnmalai and S.S. Gananamanickam, (1986)., Plant growth promoting rhizobacterialin enhancing plantgrowth and suppressing plantpathogens. Current Science 55(1), 22–25.

45. S.A. Maitlo1, R.N. Syed, M.A. Rustamani, R.D. Khuhro And A.M. Lodhi (2014)., Comparative efficacy of different fungicides against fusarium wilt of chickpea (cicer arietinum l.) Pak. J. Bot., 46(6): 2305-2312,

46. Toohey J.I., C.D. Netson and G. Krotkov, (1965)., Isolation and identification of two phenazines from a strain of Pseudomonas aureofaciens. Canadian Journal of Botany 43, 1055–1062.

47. Tronsmo A. and N. Harman, (1992)., Effect of temperature on antagonistic properties of Trichoderma species. Transactions of British Mycological Society 71, 469

48. Weller M., (1988). Biological control of soil borne plant pathogensin the rhizosphere with bacteria, Annual Reviewof Phytopathology 26, 379–407.

Received on 06.07.2018 Accepted on 05.08.2018

Asian J. Pharm. Res. 2018; 8(3): 158-166.

DOI: 10.5958/2231-5691.2018.00028.X