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首页> 外文期刊>Journal of Nematology, with Annual of Applied Nematology >Development of PrimeTime-Real-Time PGR for Species Identification of Soybean Cyst Nematode (Heterodera glycines Ichinohe, 1952)in North Carolina
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Development of PrimeTime-Real-Time PGR for Species Identification of Soybean Cyst Nematode (Heterodera glycines Ichinohe, 1952)in North Carolina

机译：开发用于在北卡罗来纳州鉴定大豆囊肿线虫（Heterodera glycines Ichinohe，1952年）的种类的PrimeTime-Real-Time PGR。

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摘要

Soybean cyst nematode (SCN) is an obligate, sedentary parasite that is a major pathogen of soybean and accounts for an estimated 1 billion dollars in production losses annually in the United States of America. This paper describes the development ofa real-time PCR method for rapid, sensitive, species-specific and accurate identification of SCN alone or on mixed populations with other nematodes in North Carolina. The 83-bp DNA fragment of PrimeTime-real-time PCR was designed based on a 477-bp-SCN-SCAR marker previously proved to be SCN-specific. A total of 44 populations including cyst forming nematodes (Heterodera glycines, H.fici, H. schachtii, H. trifolii, Cactodera weissi, Globodera tabacum, Meloidoderafloridensis and other unidentified cyst nematodes) and non-cyst forming nematodes (Ditylenchus dipsaci, Mebidogyne incognita and Xiphinema chambersi) were tested in this study, all SCN populations are tested positive and non-SCN populations negative. This assay for the detection and identification has been successfully applied for testing a single SCN cyst, a 2nd-stage-SCN juvenile, a single SCN egg, up to ten SCN cysts, a 10-fold dilution of a single 2"d-stage-SCN juvenile and 20-fold dilution of one SCN cyst. The assay is not SCN-race specific. It gave an accurate positive result when SCN is mixed with other cyst species. Also, nematode universal primers/probes for real-time PCR amplification as a nematode endogenous control to detect the presence of 18S ribosomal RNA (rRNA) gene were employed in this assay, so that a SCN-negative sample can be tested to exclude false negative. This method will be very useful for a broad range of research programs as well as the regulatory response and management of SCN in North Carolina and other region ofthe southeastern U.S.A.
机译：大豆囊肿线虫（SCN）是一种专性的，久坐的寄生虫，是大豆的主要病原体，在美国每年造成约10亿美元的生产损失。本文介绍了一种实时PCR方法的开发，该方法可用于快速，灵敏，物种特异性和准确地识别北卡罗来纳州的SCN或与其他线虫混合的种群中的SCN。 PrimeTime实时PCR的83 bp DNA片段是基于先前被证明是SCN特异性的477 bp-SCN-SCAR标记设计的。共有44个种群，包括形成囊肿的线虫（Heterodera甘氨酸，H.fici，H。schachtii，H。trifolii，Cactodera weissi，Globodera tabacum，Meloidoderafloridensis和其他未鉴定的囊肿线虫）和非囊肿的线虫（Ditylentas dipsaciyne）和Xiphinema Chambersi）在本研究中进行了测试，所有SCN群体均被测试为阳性，非SCN群体为阴性。该检测和鉴定方法已成功应用于检测单个SCN囊肿，第二阶段SCN幼虫，单个SCN卵，最多十个SCN囊肿，单个2英寸d期的10倍稀释度-SCN幼虫和一个SCN囊肿的20倍稀释液。该方法不是SCN-race特异性的。当SCN与其他囊肿种类混合时可给出准确的阳性结果。此外，线虫通用引物/探针用于实时PCR扩增作为一种线虫内源性对照，可以检测18S核糖体RNA（rRNA）基因的存在，因此可以检测SCN阴性样品以排除假阴性，这种方法对于广泛的研究将非常有用北卡罗莱纳州和美国东南部其他地区的SCN计划以及SCN的法规响应和管理

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