National Academy of Agricultural Sciences (NAAS)
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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 |
The study was undertaken to isolate the DNA of good quality from the banana wilt pathogen Fusarium oxysporum f. sp. cubense (Foc) directly from infected tissues for molecular studies. In this method, laborious and time-consuming procedures such as isolation of the pathogen and its artificial cultivation are avoided. Moreover, grinding of the fungal mycelium using liquid nitrogen is also not needed as the DNA is directly extracting from infected banana tissues. This is the first attempt to isolate Foc DNA directly from infected tissues. Extraction buffer containing 2 percent CTAB, 0.7 M NaCl, 20 mM EDTA, 50 mM Tris HCl, ß-mercaptoethanol and 1 per cent PVP was used for the isolation of fungal DNA from the infected rhizome. The concentration of DNA was 751.6 ng/µl. OD ratio λ260/ λ280 obtained was 1.89 which comesin between 1.8 to 2 indicates the purity of DNA isolated. The ITS region of 5.8S rDNA was amplified using the ITS 1F and ITS 4R primers. A single band of size approximately 580 bp was obtained as a result of PCR analysis. A tree inferred from phylogenetic analysis using Maximum Likelihood method revealed that sequence was closely related with Foc isolates which is supported by a good bootstrap value of 96 percent.
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