National Academy of Agricultural Sciences (NAAS)
|
PRINT ISSN : 2319-7692
Online ISSN : 2319-7706 Issues : 12 per year Publisher : Excellent Publishers Email : editorijcmas@gmail.com / submit@ijcmas.com Editor-in-chief: Dr.M.Prakash Index Copernicus ICV 2018: 95.39 NAAS RATING 2020: 5.38 |
1Division of Plant Pathology, FOA, main campus Chatha of SKUAST-Jammu, India
2Division of Plant Pathology, Indian Agricultural Research Institute, New Delhi, India
3Division of Biotechnology, Jammu University, Jammu, IndiaTrichoderma is one of the most important filamentous fungi used as a biocontrol agent. Because of the absence of sexual reproduction in this fungus, other methods of genetic improvement have been developed such as protoplast fusion to enhance its biocontrol potential. In order to develop an unique effective strain as a single source of vital enzyme, it was intended to integrate two biocontrol species for high chitinase production by fusing their protoplasts. Protoplasts were isolated from Trichoderma harzianum fungus using Sigma Lyticase with 0.6 M KClas osmotic stabilizer. Intra-strain T. harzianum protoplast fusion has been carried out using polyethylene glycol with STC (sorbitol, Tris – HCl, CaCl2) buffer. The maximum number of protoplasts (2.0x103/ml) was obtained from 16 h culture at pH 6,28°C for 3h. Five self-fusant strains (Th3fu1, Th3fu2, Th3fu3, Th3fu4, Th3fu5) were selected. Most of the fusants exhibited fast and vigorous mycelial growth on 2 % colloidal chitin agar. The fused protoplasts were regenerated on chitin agar selective medium. In dual culture biocontrol experiments, five selected fusants showed growth inhibition against three pathogens namely; Sclerotinia sclerotiorum, Fusarium oxysporum and Altenaria brassicicola. However, Fus 5 indicated the best ability of inhibition the growth of the three pathogens. Molecular characterization of five fusants using RAPD profile indicated the presence of novel fragments which may be due to recombination events .Results indicated a fast method to generate protoplast fusants to be used to enhance biocontrol potential intraspecifically as well as interspecifically.
Citations |
 |
 |
 |
 |
