Abstract Rice white tip nematode, Aphelenchoides besseyi, is a globally important seed-borne pathogen causing serious yield losses to rice. Since A. besseyi is transmitted via seed, its rapid identification is necessary for its management, as well as for breeding, seed certification and quarantine of rice. Several molecular markers have been developed for detection of A. besseyi. Kompetitive Allele-Specific PCR (KASP) is a novel molecular technique based on single nucleotide polymorphisms (SNP). The technique has been extensively used in studies of marker-assisted breeding, genotyping and SNP assays. Here, a novel KASP assay based on the small subunit ribosomal DNA (SSU rDNA) was developed for the identification of the rice white tip nematode. The KASP assay was successful in identifying A. besseyi isolates collected from rice fields in different locations of Thrace of Turkey and at various developmental stages. Results of the assays indicated that A. besseyi was successfully distinguished from A. ritzemabosi, A. fragariae and Robustodorus subtenuis (formerly A. subtenuis). To the best of the authors’ knowledge, this is the first approach to identify A. besseyi using a KASP assay and to investigate the use of primer extensions in KASP assay design. The developed assay can be used for accurate and rapid identification of A. besseyi.

中文翻译：

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基于 SNP 的 KASP 检测的开发和验证，用于快速鉴定 Aphelenchoides besseyi Christie，1942

摘要 水稻白尖线虫 Aphelenchoides besseyi 是一种全球重要的种子传播病原体，给水稻造成严重的产量损失。由于A. besseyi 是通过种子传播的，因此对其管理以及水稻的育种、种子认证和检疫都需要对其进行快速识别。已经开发了几种分子标记来检测 A. besseyi。竞争性等位基因特异性 PCR (KASP) 是一种基于单核苷酸多态性 (SNP) 的新型分子技术。该技术已广泛用于标记辅助育种、基因分型和 SNP 检测的研究。在这里，开发了一种基于小亚基核糖体 DNA (SSU rDNA) 的新型 KASP 检测方法，用于鉴定水稻白尖线虫。KASP 测定成功地鉴定了 A. 从土耳其色雷斯不同地点和不同发育阶段的稻田中收集的 besseyi 分离株。化验结果表明，A. besseyi 已成功与 A. ritzemabosi、A. fragariae 和 Robustodorus subtenuis（以前称为 A. subtenuis）区分开来。据作者所知，这是第一种使用 KASP 检测鉴定 A. besseyi 并研究在 KASP 检测设计中使用引物延伸的方法。开发的检测方法可用于准确快速地鉴定 A. besseyi。这是使用 KASP 检测鉴定 A. besseyi 并研究在 KASP 检测设计中使用引物延伸的第一种方法。开发的测定可用于准确和快速鉴定 A. besseyi。这是使用 KASP 检测鉴定 A. besseyi 并研究在 KASP 检测设计中使用引物延伸的第一种方法。开发的检测方法可用于准确快速地鉴定 A. besseyi。