Crop Improvement and Protection Research Site Logo
ARS Home About Us Helptop nav spacerContact Us En Espanoltop nav spacer
Printable VersionPrintable Version     E-mail this pageE-mail this page
Agricultural Research Service United States Department of Agriculture
Search
  Advanced Search
 
Programs and Projects
Subjects of Investigation
 

Title: Evaluation of molecular markers for Phytophthora ramorum detection and identification; testing for specificity using a standardized library of isolates.

Authors
item Martin, Frank
item Coffey, Mike - UC, RIVERSIDE
item Zeller, K -
item Hamelin, R - FOREST SERVICE, CANADA
item Tooley, Paul
item Garbelotto, Matteo - UC, BERKELEY
item Hughes, K - CENTRAL SCIENCE LAB. UK
item Kubisiak, T - USDA FOREST SER. MS
item Bilodeau, Guillaume -
item Levy, C -
item Blomquist, C -
item Berger, P -

Submitted to: Phytopathology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: December 29, 2008
Publication Date: April 1, 2009
Citation: Martin, F.N., Coffey, M., Berger, P., Hamelin, R.C., Tooley, P.W., Garbelotto, M., Hughes, K., Kubisiak, T. 2009. Evaluation of molecular markers for Phytophthora ramorum detection and identification; testing for specificity using a standardized library of isolates. Phytopathology 98:390-403.

Interpretive Summary: This experimentation evaluates different molecular detection methods for identification of the pathogen Phytophthora ramorum, which is responsible for sudden oak death.

Technical Abstract: A number of molecular techniques have been developed for detection of Phytophthora ramorum from infected tissue. These have been based on spacer regions (the rDNA ITS region, the spacer region between the cox I and II gene) or specific genes (beta tubulin, elicitin) and have been configured for use by both conventional and real-time PCR. Techniques based on PCR-RFLP of the cox 1 and 2 gene cluster or single strand conformational polymorphism of the ITS region also have been proposed for isolate identification. In an effort to make a uniform comparison of specificity among the techniques a standard library of DNA was extracted from 317 isolates representing 60 described species and 22 isolates with ambiguous taxonomic classification. Aliquots were adjusted to 10ng/µl, coded and sent blind to all participants (a total of 457 samples were sent). For all extracts the ITS region and cox2 gene were also amplified and sequenced to confirm the identity of the samples. A total of 12 marker systems were evaluated (conventional PCR: cox 1 and 2 spacer, ITS; real-time PCR: cox 1 and 2 spacer, ITS, beta tubulin, elicitin), many by the labs in which they were developed. The results of these evaluations and their implications for effective diagnostics of P. ramorum were discussed.

   
 
 
Last Modified: 05/26/2013
ARS Home | USDA.gov | Site Map | Policies and Links 
FOIA | Accessibility Statement | Privacy Policy | Nondiscrimination Statement | Information Quality | USA.gov | White House