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ARS Home » Northeast Area » Beltsville, Maryland (BARC) » Beltsville Agricultural Research Center » National Germplasm Resources Laboratory » Research » Publications at this Location » Publication #304330

Title: Use of nested PCR to detect Ceratocystis fagacearum in sapwood of diseased northern oak species

item YOUNG, ANNA - University Of Minnesota
item JUZWIK, JENNIFER - Us Forest Service (FS)
item Mollov, Dimitre

Submitted to: American Phytopathological Society Annual Meeting
Publication Type: Abstract Only
Publication Acceptance Date: 5/1/2014
Publication Date: 8/6/2014
Citation: Young, A., Juzwik, J., Mollov, D.S. 2014. Use of nested PCR to detect Ceratocystis fagacearum in sapwood of diseased northern oak species. American Phytopathological Society Annual Meeting. Phytopathology 104:S3.132.

Interpretive Summary:

Technical Abstract: Early and accurate diagnosis of oak wilt, caused by Ceratocystis fagacearum (Cf), is important when disease control action is planned. When laboratory diagnosis is needed, standard isolation protocols that are used rely on high quality samples and require > 14 days for incubation. Use of a nested PCR protocol to detect Cf in diseased sapwood shavings was compared to detection success using published isolation methods. Assays were performed on samples from oak species common in the Lake States. Four subsamples of three branches collected from each tree were tested. In actively wilting red oaks, 97% of the subsamples from nine trees were positive for Cf using PCR compared to 82% using isolation techniques. For wilting branches from eight bur oaks, 92% of subsamples were positive for Cf versus 67% based on isolation. In eight white oaks, 93% of subsamples were positive for Cf using PCR compared to 47% with isolation. The fungus was not detected by either technique in subsamples of branches obtained from healthy oaks (controls). For bur and white oak branches dead for at least 1 year, Cf was detected using PCR (55 and 87% of subsamples, respectively), but was not detected by isolation. Similarly, only the PCR assay detected the pathogen in sapwood samples underlying remnant Cf sporulation mats on three main stem locations of seven trees. These findings are of particular interest to diagnostic laboratories that process disease-suspect samples in the region.