Exploitation of abiotic stress tolerant strains of trichoderma spp. for the management of soil borne fungal pathogens

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Date
2018
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Department of Plant Pathology, College of Horticulture, Vellanikkara
Abstract
Soil borne phytopathogenic fungi are known to cause severe yield loss of several crops. To mitigate these crop ailments, farmers mostly rely on chemical methods of disease control such as fungicides and other pesticides which are deleterious to the environment. To date, biological control is an ecofriendly approach for the effective management of crop diseases. The fungi belonging to the genus Trichoderma are one among the most exploited biocontrol agents in the field of agriculture. However, the performance of Trichoderma spp. gets reduced when it is exposed to abiotic stressed conditions such as drought, high temperature, salinity, acidity and fungicides. Hence, this study was proposed to identify and exploit stress tolerant isolates of Trichoderma spp. with antagonistic potential in Kerala. Intensive soil sampling surveys were conducted across different stressed ecosystems of Kerala viz., Palakkad, Alappuzha, Vytilla, Kumarakom, Wayanad and Thrissur for the isolation and enumeration of native Trichoderma spp. A total of 24 isolates were obtained from 52 soil samples collected from different locations. Based on the number of Trichoderma spp. obtained from each district, they were serially numbered and abbreviated according to the name of the location. Accordingly, PAT 1 to PAT 6 represents number of isolates of Trichoderma spp. from Palakkad district, ALT 1 to ALT 3 from Alappuzha, VYT 1 and VYT 2 from Ernakulam district, KUT 1 from Kottayam district, WAT 1 to WAT 8 from Wayanad and THT 1 to THT 4 from Thrissur district. Cultural and morphological identification of these isolates were carried out under in vitro conditions. Isolates of Trichoderma spp. were subjected to in vitro screening for abiotic stress tolerance such as high temperature, drought, acidity, salinity and also to test their sensitivity towards copper fungicides. The isolates PAT6 and WAT2 were found as thermotolerant, VYT2 and ALT 1 as drought tolerant, ALT 3 and ALT 1 as acid tolerant and saline tolerant and the isolates ALT1, ALT3 and PAT 1 as copper fungicide tolerant. The selected six isolates were further subjected to biochemical tests and the study showed that the isolates VYT 2, ALT 3 and ALT 1 showed highest cellulase, β- 1, 3 glucanase and protease activity. Likewise, isolates PAT 1, ALT 1 and ALT 3 were found as best producers of ACC deaminase and PAT 1 and ALT 1 as the best cytokinin producer. The best performing isolates (ALT 1, ALT 3, PAT 1, PAT 6 and VYT 2) after enzyme study were subjected to dual culture experiment with five major soil borne pathogens to test their antagonistic potential. The isolates ALT 3, ALT 1, PAT 6 and PAT 1 showed more than 70 per cent inhibition of R. solani whereas, isolates ALT1 and VYT 2 showed only 57.78 and 55 per cent inhibition of S. rolfsii respectively. However, no significant difference was noticed among the isolates when grown against F. solani. Cent per cent inhibition of P. aphanidermatum was noticed with Trichoderma isolates PAT 1 and ALT 1. All five isolates showed 100 per cent inhibition on the growth of pathogen, P. capsici. Among the five, four isolates viz., ALT 1, ALT 3, PAT 1 and PAT 6 with best antagonistic potential were subjected to molecular characterization at Rajiv Gandhi Centre for Biotechnology, Thiruvananthapuram and the isolates showed homology with the nucleotide sequence of Trichoderma asperellum. A pot culture experiment was laid with the selected isolates viz., ALT 1, ALT 3, PAT 1 and PAT 6 to test the growth promotion of cowpea and biocontrol efficacy against Rhizoctonia solani. It was observed that the isolate PAT 6 coated seeds showed 100 per cent germination and also recorded better biometric characters and yield. Moreover, the lowest per cent disease incidence of 11.11 per cent was only recorded with both the isolates ALT 3 and PAT 6. Thus, the study has enlightened our knowledge on the existence of abiotic stress tolerant isolates of T. asperellum which can be employed in future for the biocontrol of soil borne pathogens in such conditions.
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