Conventional and PCR Detection of Aphelenchoides fragariae in Diverse Ornamental Host Plant Species |
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| Authors: | Jamie L McCuiston Laura C Hudson Sergei A Subbotin Eric L Davis and Colleen Y Warfield |
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| Institution: | 1Graduate Students, Professor and Assistant Professor, Department of Plant Pathology, NC State University, Raleigh, NC 27695 2Plant Pest Diagnostic Center, California Department of Food and Agriculture, 3294 Meadowview Road, Sacramento, CA 95832 3Current Address: University of California Cooperative Extension, 80 Stone Pine Road, Suite 100, Half Moon Bay, CA 94019 |
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| Abstract: | A PCR-based diagnostic assay was developed for early detection and identification of Aphelenchoides fragariae directly in host plant tissues using the species-specific primers AFragFl and AFragRl that amplify a 169-bp fragment in the internal transcribed spacer (ITS1) region of ribosomal DNA. These species-specific primers did not amplify DNA from Aphelenchoides besseyi or Aphelenchoides ritzemabosi. The PCR assay was sensitive, detecting a single nematode in a background of plant tissue extract. The assay accurately detected A. fragariae in more than 100 naturally infected, ornamental plant samples collected in North Carolina nurseries, garden centers and landscapes, including 50 plant species not previously reported as hosts of Aphelenchoides spp. The detection sensitivity of the PCR-based assay was higher for infected yet asymptomatic plants when compared to the traditional, water extraction method for Aphelenchoides spp. detection. The utility of using NaOH extraction for rapid preparation of total DNA from plant samples infected with A. fragariae was demonstrated. |
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| Keywords: | Aphelenchoides fragariae detection diagnosis foliar nematode ITS1 method NaOH ornamental host PCR rDNA |
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