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United States Department of Agriculture

Agricultural Research Service

Title: The Use of Mitochondrial DNA for Clarifying Phylogenetic Relationships and Isolate Identification with the Genus Phytophthora

Authors
item Martin, Frank
item Tooley, Paul

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: March 22, 2005
Publication Date: August 20, 2005
Citation: Martin, F.N., Tooley, P.W. 2005. The use of mitochondrial dna for clarifying phylogenetic relationships and isolate identification with the genus phytophthora. Meeting Abstract. No page numbers.

Technical Abstract: The mitochondrial genome in Phytophthora has provided additional tools for phylogentic analysis (cox 1 and 2 genes) of the genus that allows for greater resolution among closely related species than the more commonly used rDNA-ITS region. In addition, a molecular marker system for detection of the pathogen in infected tissue was developed from mtDNA that uses a primer pair that is specific for Phytophthora spp. multiplexed with a plant primer pair that serves as a positive control. A second round amplification with nested species-specific primers was used to detect specific species. This marker system was most widely tested with P. ramorum, but has been found to work on a range of other species as well. The species-specific marker system can be used with traditional PCR as well as real-time PCR (SYBR Green and TaqMan). An alternative approach for isolate identification was developed using RFLP analysis of another PCR amplified region to accurately identify isolates to a species level. In evaluation of isolates representing 31 species isolates were consistently identified to the correct species, including those not clearly differentiated by RFLP analysis of the rDNA-ITS region.

Last Modified: 7/31/2014
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