从土壤中提取的南方根结线虫、爪哇根结线虫和花生根结线虫的灵敏PCR检测
Sensitive PCR Detection of Meloidogyne arenaria, M. incognita, and M. javanica Extracted from Soil.
作者信息
Qiu Jinya Jack, Westerdahl Becky B, Anderson Cindy, Williamson Valerie M
机构信息
Department of Nematology, University of California, One Shields Avenue, Davis, CA 95616.
出版信息
J Nematol. 2006 Dec;38(4):434-41.
We have developed a simple PCR assay protocol for detection of the root-knot nematode (RKN) species Meloidogyne arenaria, M. incognita, and M. javanica extracted from soil. Nematodes are extracted from soil using Baermann funnels and centrifugal flotation. The nematode-containing fraction is then digested with proteinase K, and a PCR assay is carried out with primers specific for this group of RKN and with universal primers spanning the ITS of rRNA genes. The presence of RKN J2 can be detected among large numbers of other plant-parasitic and free-living nematodes. The procedure was tested with several soil types and crops from different locations and was found to be sensitive and accurate. Analysis of unknowns and spiked soil samples indicated that detection sensitivity was the same as or higher than by microscopic examination.
我们已经开发出一种简单的聚合酶链式反应（PCR）检测方案，用于检测从土壤中提取的根结线虫（RKN）种类，即南方根结线虫、爪哇根结线虫和花生根结线虫。使用贝曼漏斗法和离心浮选法从土壤中提取线虫。然后用蛋白酶K消化含线虫部分，并使用针对该组根结线虫的特异性引物和跨越核糖体RNA基因内转录间隔区（ITS）的通用引物进行PCR检测。在大量其他植物寄生线虫和自由生活线虫中能够检测到根结线虫二龄幼虫（J2）。该程序在来自不同地点的几种土壤类型和作物上进行了测试，结果表明其灵敏且准确。对未知样品和加标土壤样品的分析表明，检测灵敏度与显微镜检查相同或更高。
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