Direct extraction and purification of soil microbial total DNA and molecular detection of soil-borne pathogen Fusarium oxysporum causal agent of tomato vascular wilt

Document Type : Research Paper

Authors

1 Associate Professor, Department of Agricultural Biotechnology, Payame Noor University, Tehran, Iran

2 M.Sc. of Biotechnology, Department of Agricultural Biotechnology, Payame Noor University, Tehran, Iran

3 . Ph.D. of Plant Pathology, Department of Plant Pathology, College of Agriculture and Natural Resources, Science and Research Branch, Islamic Azad University, Tehran, Iran

Abstract

Molecular analysis is often required in many genome extracted from various tissues. In much molecular analysis of genetic studies of microbial communities, often require direct extraction of DNA from soil. In this study, after careful study of the various methods proposed three methods (chemical enzyme), (chemi-mechanical) and (chemical mechanical enzymatic), in order to extract DNA from soil, a separate project was studied to most identify different methods of extraction. The comparison between the same conditions, a significant difference in the DNA product of the average concentration of 2/32 to 6/1 ng per µl DNA was diluted and also on the basis of information obtained from Nanodrop and electrophoresis device and review of the DNA extracted protein product DNA, methods (chemical, mechanical, enzymatic) better performance at higher concentrations, remove humic acid proteins and other contaminants was superior to other methods. The molecular detection of pathogens that extraction is important applications; the aim of this study is molecular detection of pathogen Fusarium oxysporum f. sp lycopersici and Fusarium oxysporum f. sp. radicis lycopersici of soil microbial community by molecular techniques to identify the special primer evaluated. Molecular detection results showed that there is tomato Fusarium infection in soil samples S1.

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Crop Biotechnology
Volume 5, Issue 2 - Serial Number 12
February 2016
Pages 55-66

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