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A species-specific PCR assay for the newly observed root lesion nematode, Pratylenchus vulnus, in Taiwan

Yu-po Lin: National Taiwan university

<div>The root lesion nematode <em>Pratylenchus sp.</em> is the third most agriculture- impacting plant parasitic nematode genus. On March 2017, <em>Pratylenchus vulnus</em> was extracted from a strawberry field soil in Taiwan. The nematode was identified both morphologically and molecularly. Measurements were taken under the view of compound microscopic observation. Molecular analysis were done on the sequences of the cytochrome oxidase subunit I (COI) gene of mitochondria DNA (mtDNA) and multiple regions of the ribosome gene, 18S, ITS region, and the D2/D3 region of 28S. It was the first record of <em>P. vulnus</em> in Taiwan. A species-specific PCR detection assay targeting a 138-bp long region of the mtDNA COI gene of <em>P. vulnus</em> was then developed for rapid detection and identification. The detection limit of the assay is 10<sup>4</sup> copy per µl when examined with cloned target DNA. The assay is highly specific and can differentiate the target from the closely related and easily misdiagnosed spscies, such as <em>Pratylenchus speijeri, Pratylenchus coffeae, Pratylenchus kumamotoensis,</em> and other common plant parasitic nematodes including <em>Helicotylenchus sp.,</em><em> Rhadinaphelenchus sp., Radopholus sp.,Tylenchorhynchus sp. Ditylenchus sp.,</em> and <em>Bursaphelenchus xylophilus.</em></p> <p style="text-align:left;"><em> </em></p> <p style="text-align:left;">Key word:<em> Pratylenchus vulnus</em>; mtDNA; PCR</div>