عنوان مقاله [English]
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.
Ahmadikhah A (2012) Genetic Engineering. Gorgan University of Agriculture Sciences & Natural resources, Iran.
Darvishnia M, Dehghani A (2015) Diseases of vegetable crops in Iran and their integrated Management, Lorestan University, Iran.
Ebrahimi MA, Tohifar M, Ahmadinik I (2014) Principles of Genetic Engineering. Payame Noor University, Iran.
Burgmann H, Pesaro M, Widmer F, Zeyer J (2001) A strategy for optimizing quality and quantity of DNA extracted from soil. J. Microbial. Methods 45: 7-20.
Courtois S, Frostegard AS, Goransson P (2001) Quantification of bacterial subgroups in soil: comparison of DNA extracted directly from soil or from cells previously released by density gradient centrifugation. Environ. Microbiol. 7:431- 439.
De Lipthay JR, Enzinger C, Johnsen K, Aamaand J, Sorensen SJ (2004) Impact of DNA extraction method on bacterial community composition measured by denaturing gradient gel electrophoresis. Soil Biol. Biochem. 36: 1607-1614.
Hart ML, Meyer A, Johnson PJ, Ericsson AC (2015) Comparative evaluation of DNA extraction methods from feces of multiple host species for downstream next-generation sequencing. PLoS ONE 10(11): e0143334. doi:10.1371/journal.pone.0143334.
Hirano Y, Arie T (2006) PCR-based differentiation of Fusarium oxysporum ff. sp. lycopersici and radicis-lycopersici and races of F. oxysporum f. sp. lycopersici. J. Gen. Plant Path. 72: 273-283.
Islam MR, Trivedi P, Palaniappan P, Reddy MST (2009) Evaluating the effect of fertilizer application on soil microbial community structure in rice based cropping system using fatty acid methyl esters (FAME) analysis World J. Microbiol. Biotechnol. 25: 1115-1117.
Knietsch A, Waschkowitz T, Bowien S, Henne A, Daniel R (2003) Metagenomics of complex microbial consortia derived from different soils as sources for novel genes conferring formation of carbonyls from short-chain polyols on Escherichia coli. J. Mol. Microbiol. Biotechnol. 5: 46-56.
Liesack W, Weyland H, Stackebrandt E (1991) Potential risks of gene amplification by PCR as determined by 16s rDNA analysis of a mixed–culture of strict barophilic bacteria: Microb. Ecol. 21: 191-198.
Luna GM, Dell Anno A, Danaovaro R (2006) DNA extraction procedure: a critical issue for bacterial assessment in marine sediments. Environ. Microbiol. 8: 308-320.
Luo H, Qi H, Xue K, Zhang H (2003) A preliminary application of PCR–DGGE to study microbial diversity in soil. Acta Ecologica Sini. 23: 1570-1575.
Robe P, Nalin R, Capellano C, Vogel TM (2003) Extraction of DNA from soil. Eur. J. Soil Biol. 39: 183-190.
Roose-Amsaleg CL, Garnier-Sillam E, Harry M (2001) Extraction and purification of microbial DNA from soil and sediment samples. Appl. Soil Ecol. 18: 47-60.
Torsvik V, Ovreas L (2002) Microbial diversity and function in soil: from genes to ecosystems. Curr. Opin. Microbiol. 5: 240-245.
Yeates C, Gilling MR, Davision AD, Altavilla N, Veal DA (1998) Methods for microbial DNA extraction from soil for PCR amplification. Biol. Proc. Online 1: 40-47.
Zhou JZ, Mary AB, James MT (1996) DNA recovery from soils of diverse composition. Appl. Environ. Microbiol. 62: 316-322.
Zhang R, Cao H, Cui Z, Li S, Fan, B. 2003. Extraction and purification of soil microbial total DNA. Acta Microbiol. Sini . 43: 276-282.