Study on PCR rapid molecular detection technique of Meloidogyne vitis

文献类型： 外文期刊

第一作者： Yang, Yan-Mei

作者： Yang, Yan-Mei;Du, Xia;Dong, Ye;Hu, Xian-Qi;Yang, Yan-Mei;Du, Xia;Dong, Ye;Hu, Xian-Qi;Liu, Pei;Li, Hong-Mei;Peng, Huan

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关键词： Meloidogyne vitis; PCR rapid detection; reliability; specificity; sensitivity

期刊名称：JOURNAL OF INTEGRATIVE AGRICULTURE （ 影响因子：4.384； 五年影响因子：4.021 ）

ISSN： 2095-3119

年卷期： 2022 年 21 卷 11 期

页码：

收录情况： SCI

摘要： Meloidogyne vitis is a new root-knot nematode parasitic on grape root in Yunnan Province, China. In order to establish a rapid, reliable and specific molecular detection method for M. vitis, the species-specific primers were designed with rDNA-ITS (ribosomal DNA internal transcribed spacer) gene fragment as the target. The reaction system was optimized and the reliability, specificity and sensitivity of primer were testified, therefore, a rapid PCR detection method for M. vitis was established. The result showed that the optimal annealing temperature of the primers was 53 degrees C, which was suitable for the detection of different life stages of M. vitis. Specificity test showed that the specific fragment size of 174 bp was obtained from M. vitis, but other five non-target nematodes did not have any amplification bands, thus effectively distinguish M. vitis and the other five species, and could specifically detect the M. vitis from mixed populations. Sensitivity test showed that this PCR technique could detect the DNA of a single second-stage juvenile (J2) and 10-4 female. Futhermore, this PCR technique could be used to detect directly M. vitis from soil samples. The rapid, sensitive and specific PCR molecular detection technique could be used for the direct identification of a single J2 of M. vitis and the detection of M. vitis in mixed nematode populations and the detection of two J2s or one male in 0.5 g soil samples, which will provide technical support for the investigation of the occurrence and damage of M. vitis and the formulation of efficient green control strategies.

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