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

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https://ir.library.oregonstate.edu/concern/articles/s7526f12k

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  • 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.
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  • 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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  • 97
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  • 12
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  • This research was supported by the Idaho Wheat Commission, Oregon Wheat Commission, and Washington Grains Commission; Oregon State University, Agricultural Research Foundation project ARF 7136A; and a subcontract between Oregon State University and the United States Department of Agriculture–Agricultural Research Service (SCA 58-5348-9-100, “Control of Root Diseases of Wheat and Barley”).
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  • description.provenance : Submitted by Erin Clark (erin.clark@oregonstate.edu) on 2014-03-18T16:21:03Z No. of bitstreams: 1 YanGuipingCropSoilScienceSpecies-SpecificPCR.pdf: 358570 bytes, checksum: a356386df2ee398fb538b07fc79c1a4f (MD5)
  • description.provenance : Approved for entry into archive by Erin Clark(erin.clark@oregonstate.edu) on 2014-03-18T16:21:39Z (GMT) No. of bitstreams: 1 YanGuipingCropSoilScienceSpecies-SpecificPCR.pdf: 358570 bytes, checksum: a356386df2ee398fb538b07fc79c1a4f (MD5)
  • description.provenance : Made available in DSpace on 2014-03-18T16:21:39Z (GMT). No. of bitstreams: 1 YanGuipingCropSoilScienceSpecies-SpecificPCR.pdf: 358570 bytes, checksum: a356386df2ee398fb538b07fc79c1a4f (MD5) Previous issue date: 2013-12

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