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Synaptic guidepost protein (syg-2) gene-specific primers for detecting Bursaphelenchus xylophilus

Xizhuo Wang: Research Institute of Forest Ecology, Environment and Protection,CAF

<div>Pine wilt disease (PWD) caused by pine wood nematode (PWN) is a worldwide quarantine disease that is detrimental to pine trees. A stable and highly specific <em>Bursaphelenchus xylophilus</em> gene helps to accurately diagnose diseased pine trees. This study standardizes the PCR and loop-mediated isothermal amplification (LAMP) methods using newly designed primer sequences of the syg-2 gene, which encodes the synaptogenesis protein syg-2, was involved in many major physiological functions and was conserved in <em>B. xylophilus</em>, the region showed 46% dissimilarity (i.e., 211 in 398bp is identical) between <em>B. xylophilus</em> and <em>B. mucronatus</em>, while this dissimilarity in Nuclear ribosomal internal transcribed spacer (ITS) region was 20 %. Our experimental results suggest that the syg-2 gene makes a good distinction between B. xylophilus and other nematodes. However,when using the ITS region design primers for LAMP detection, <em>B. frandulentus</em> and <em>B.doui </em>could be detected in about 50 minutes, and<em> B. mucronatus</em> began to be detected in about 90 minutes. Therefore, syg-2 gene can be used as a new target gene for detection of PWN.</div>