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Species-Specific PCR Assays for Differentiating Heterodera filipjevi and H. avenae

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Title Species-Specific PCR Assays for Differentiating Heterodera filipjevi and H. avenae
Names Yan, Guiping (creator)
Smiley, Richard W. (creator)
Okubara, Patricia A. (creator)
Skantar, Andrea M. (creator)
Date Issued 2013-12 (iso8601)
Note To the best of our knowledge, one or more authors of this paper were federal employees when contributing to this work. This is the publisher’s final pdf. The published article is copyrighted by the American Phytopathological Society and can be found at: http://apsjournals.apsnet.org/loi/pdis.
Abstract Heterodera avenae and H. filipjevi are economically important cyst
nematodes that restrict production of cereal crops in the Pacific Northwest
United States and elsewhere in the world. Identification of these
two species is critical for recommending and implementing effective
management practices. Primers were designed from the internal transcribed
spacer (ITS) regions of H. avenae and H. filipjevi ribosomal
DNA. The primers were highly specific when examined on target isolates
but did not amplify DNA from nontarget Heterodera, Globodera,
Meloidogyne, Pratylenchus, and other nematode species tested. Polymerase
chain reaction (PCR) and amplification conditions were established, and H. avenae and H. filipjevi were clearly distinguished by
PCR fragments of 242 and 170 bp, respectively. Robust PCR amplification
was achieved with DNA extracted from a single egg or second-stage
juvenile (J2) using a laboratory-made worm lysis buffer, and
DNA from 0.5 egg or J2 using a commercial kit. The PCR assays were
successfully employed for differentiation of H. filipjevi and H. avenae
populations collected from eight locations in three Pacific Northwest
states. This is the first report of a species-specific ITS PCR assay to
detect and identify H. filipjevi. The assays for both species will enhance
diagnosis of cereal cyst nematode species in infested fields.
Genre Article
Identifier Yan, G., Smiley, R. W., Okubara, P. A., & Skantar, A. M. (2013). Species-Specific PCR Assays for Differentiating Heterodera filipjevi and H. avenae. Plant Disease, 97(12), 1611-1619. doi:10.1094/PDIS-01-13-0064-RE

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