K.S. Sayyed, K.T. Apet, K.B. Rudrappa and V.B. Bhujabal

Department of Plant Pathology, Vasantrao Naik Marathwada Krishi Vidyapeeth, Parbhani- 431402, Maharashtra, India.

Abstract

Effective native isolates of Trichoderma spp. collected from the rhizosphere of different plant growing in the field. Twenty seven isolates of Trichoderma spp 13, T. viride (Pers.), 9, T. harzianum (Rifai), 2, T. longibrachiatum (Rifai), 1, T. koningii (Oudem) and 2, Gliocladium virens were identified. The antagonistic potential of the isolates as evaluated through dual culture plate using three soil borne pathogens of wilt of Safflower, Chickpea and Pigeonpea respectively, T. harzianum Jalna isolate was found highly antagonistic compared to T. viride isolates as it inhibited mycelia growth of Fusarium oxysporum f.sp. carthami, F. oxysporum f.sp. ciceri and F. oxysporum f.sp. udum by 78.06 %, 89.99 % and 82.49 per cent respectively after six days of in vitro incubation.

Keyword: Trichoderma spp., Rhizosphere, Biocontrol, Fusarium sp., Safflower, Pigeonpea and Chickpea.

Introduction

Trichoderma species are used as biocontrol agent in agriculture. Trichoderma, a genus of asexually reproducing saprophytic fungi, frequently present in nearly all temperate and tropical soils, decaying plant tissues and root ecosystems. The strains of Trichoderma spp. are strong opportunistic invaders, fast growing, prolific producers of spores and powerful antibiotic producers. The antagonism of Trichoderma involves several mechanisms, such as competition for nutrient antibiosis and production of fungal cell wall degrading enzymes. The mycoparasitic ability of Trichoderma species against plant pathogenic filamentous fungi allows for development of biocontrol strategies (Benitez et al., 2004).
The Fusarium spp. is one of the most important soil borne fungi which causes wilt diseases in several field crops like chickpea, pigeonpea and safflower. Yield losses in different field crops due to Fusarium are upto tune of 90 per cent (Singh and Dahiya 1973).

Materials and Methods

Sample Collection
A total of 35 samples were randomly collected from different sites in Jalna and Parbhani district from August to Sept. 2014. The soil samples were taken from a depth of 05 to 15 cm around the rhizosphere area of fruit plants, cereal, vegetable, pulses and oilseed crops.
Isolation of Trichoderma spp. from rhizospheric soils
Serial dilution technique was used to isolate the Trichoderma from the samples collected. The collected samples were air dried in shade. These samples were finely ground before serial dilutions. Trichoderma Selectve Medium (TSM) was used for isolation. Isolation was carried out under aseptic conditions of laminar air flow in aseptic condition. Isolation were carried out by serial dilution and plating method. The culture of bioagents Trichoderma spp isolated from different region was identified on the basis of morphological and cultural characteristics.
In vitro evaluation for antagonistic potential
Twenty seven Trichoderma spp isolates, thirteen T. viride (Pers.), nine T. harzianum (Rifai), two T. longibrachiatum (Rifai), one T. koningii (Oudem) and two G. virens were evaluated in vitro against Fusarium oxysporum f.sp. carthami, F. oxysporum f.sp. ciceri and F. oxysporum f.sp. udum by employing dual culture technique by Dennis and Webster (1971). Seven days old cultures of bioagents and test fungus grown on agar media were used for the study. Disc (5 mm diameter) of PDA along with culture grown of the test fungus and bioagents were cut out with sterilized cork borer. Then two cultures disc, one each of the test fungus and bioagents were placed at equidistance and exactly opposite with each other on solidified PDA medium in plates under aseptic conditions and plates were incubated at 28±2o C. plates inoculated with culture disc of test fungus were maintained and untreated control. Observations on linear mycelia of 24 hours and continued till untreated control plate was fully covered with mycelia growth of the test fungus. Per cent inhibition of the test fungus over untreated control was calculated by applying the formula given by Arora and Upadhyay (1978).

After incubation radial growth was calculated by applying the formula (Vincent, 1927)
Per cent inhibition (I) = C-T/C x 100

Where,
C = Growth of test fungus in control in mm.
T = Growth of test fungus in treatment in mm.

Result and Discussion

In vitro evaluation of bioagents
In vitro bioefficacy of different Trichoderma isolates against F. oxysporum f. sp. carthami.
The results (Table1) obtained on mycelial growth and inhibition of Fusarium oxysporum f. sp. carthami (Klisiewicz and Houston) with thirteen T. viride, nine T. harzianum, two T. longibrachiatum, one T. koningii, two G. virens isolates antagonists. It was observed that treatment T7 (T. harzianum) isolate obtained from Jalna strain showed highest zone of inhibition i.e. 78.06 per cent followed by T17 (T. harzianum) Parbhani strain i.e. 77.50 and T2 (T. harzianum) Jalna strain i.e. 76.66 per cent.
Results of the present study on effect of bioagents on mycelia growth of F. oxysporum f. sp. carthami are in consonance with those reported earlier by several workers. Bioagents viz., Trichoderma viride, T. harzianum, T. longibrachiatum, T. koningii, and G. virens were reported earlier to cause significant inhibition of mycelial growth of F. oxysporum f. sp. carthami by several workers (Behare et al., 2002; Singh et al., 2003; Waghmare and Datar, 2007 and Waghmare and Datar, 2009).
In vitro bioefficacy of different Trichoderma isolates against F. oxysporum f. sp. ciceri.
The results (Table 2) obtained on mycelial growth and inhibition of Fusarium oxysporum f. sp. ciceri (Padwick) with thirteen T. viride, nine T. harzianum, two T. longibrachiatum, one T. koningii, two G. virens isolates antagonists. It was observed that treatment T3 (T. harzianum) isolate obtained from Jalna strain showed highest zone of inhibition i.e. 89.99 per cent followed by T27 (T. viride) Parbhani strain i.e. 87.21 and T7 (T. harzianum) Jalna strain i.e. 86.94 per cent.
Results of the present study on effect of bioagents on mycelia growth of F.oxysporum f. sp. ciceri are in consonance with those reported earlier by several workers. Bioagents viz., Trichoderma viride, T. harzianum, T. longibrachiatum, T. koningii, and G. virens were reported earlier to cause significant inhibition of mycelial growth of F.oxysporum f. sp. ciceri by several workers ( Srivastava and Mall, 2008; Patil et al., 2010; Rajput et.al., 2010 and Yadav and Anadani , 2013).
In vitro bioefficacy of different Trichoderma isolates against F. oxysporum f. sp. udum.
The results (Table 3) obtained on mycelia growth and inhibition of Fusarium oxysporum f. sp. udum (Butler) with thirteen T. viride, nine T. harzianum, two T. longibrachiatum, one T. koningii, two G. virens isolates antagonists. It was observed that treatment T12 (T. harzianum) isolate obtained from Jalna strain showed highest zone of inhibition i.e. 82.49 per cent followed by T5 and T10 (T. harzianum) Jalna strain i.e. 81.10 and T24 (T. viride) Parbhani strain i.e. 80.83 per cent.
Results of the present study on effect of bioagents on mycelial growth of F. oxysporum f. sp. udum are in consonance with those reported earlier by several workers. Bioagents viz., Trichoderma viride, T. harzianum, T. longibrachiatum, T. koningii, and G. virens were reported earlier to cause significant inhibition of mycelial growth of F. oxysporum f. sp. udum by several workers (Pandey and Upadhay, 2000; Patel et al., 2011; Ranjana and Chhetry, 2012).

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