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International Journal of Current Microbiology and Applied Sciences (IJCMAS)
IJCMAS is now DOI (CrossRef) registered Research Journal. The DOIs are assigned to all published IJCMAS Articles.
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National Academy of Agricultural Sciences (NAAS)
NAAS Score: *5.38 (2019)
[Effective from January 1, 2019]
For more details click here

ICV 2018: 95.39
Index Copernicus ICI Journals Master List 2017 - IJCMAS--ICV 2018: 95.39
For more details click here

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Original Research Articles

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

Int.J.Curr.Microbiol.App.Sci.2020.9(6): 2587-2593
DOI: https://doi.org/10.20546/ijcmas.2020.906.315


Simple and Rapid Method for Extraction of Good Quality Fungal DNA Directly from Fusarium Wilt Infected Banana and Molecular Characterization of the Pathogen
N. P. Lishma1*, K. Anita Cherian1 and Deepu Mathew2
1Department of Plant Pathology,
2Department of Plant Biotechnology, College of Horticulture, Kerala Agricultural University, Thrissur, 680 656, India
*Corresponding author
Abstract:

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.


Keywords: Foc,DNA isolation,PCR, sequencing, phylogenetic analysis
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How to cite this article:

Lishma. N. P., K. Anita Cherian and Deepu Mathew. 2020. Simple and Rapid Method for Extraction of Good Quality Fungal DNA Directly from Fusarium Wilt Infected Banana and Molecular Characterization of the Pathogen.Int.J.Curr.Microbiol.App.Sci. 9(6): 2587-2593. doi: https://doi.org/10.20546/ijcmas.2020.906.315