Development of PCR-based diagnostics for soil borne plant pathogens

Autor: R. N. Nazar, X. Hu, Tatiana Volossiouk, E. J. Robb, Shin Woo Lee
Rok vydání: 1997
Předmět:
Zdroj: Journal of Plant Biology. 40:176-181
ISSN: 1867-0725
1226-9239
Popis: We believe our success with a PCR-based assay forVerticillium in plant tissue and soil samples represents a good model for other developments of this type. The universal presence of high copy ribosomal RNA genes in all organisms makes these genes especially attractive targets for gene isolation and PCR amplification. A sample dilution strategy permits the direct extraction of DNA from either soil or tissue samples without further purification, a feature which permits low cost and wide scale applications. The use of internal control templates largely eliminates the possibility of false negatives and permits quantitative analysis, something which is usually very inaccurate or impossible with traditional assays. Finally the careful preparation of PCR reagents and the use of negative controls eliminate the possibility of false positives, a careful selection of primer sequence and anneall ing conditions permits the differentiation of even closely related species. With an aim at rapid, low cost, wide scale testing a number of compromises have been included in the approaches with satisfactory results. Recent applications to nematode diagnostics appear to support the general application of our approaches to many important crop pathogens.
Databáze: OpenAIRE
Externí odkaz: