Heterodera filipjevi许昌群体SCAR标记的建立

菲利普孢囊线虫Heterodera filipjevi是禾谷类作物上重要的病原线虫之一,严重影响禾谷类作物的产量和品质。本课题组前期研究发现菲利普孢囊线虫许昌群体是一个新的致病型,其在小麦上的致病力强于其他多个群体,危害更为严重。本研究旨在建立菲利普孢囊线虫许昌群体的快速、准确的分子检测体系,为Heterodera filipjevi许昌群体的监测和防控及抗病品种的选育利用奠定基础。本研究采用随机扩增多态性DNA(RAPD)和序列特征扩增区域(SCAR)的方法,对黄淮麦区5个重要小麦孢囊线虫致病型共9个线虫群体进行RAPD分析和SCAR标记转化,并通过增加除黄淮麦区外的线虫群体验证所获得的致病型相关分子标记的特异性和有效性。本研究共筛选了331条RAPD引物,筛选出2个菲利普孢囊线虫许昌群体相关的RAPD标记,引物S86可以扩增出1条约550 bp的多态性片段,引物S178可以扩增出1条约1 200 bp的多态性片段,并将这2个RAPD标记成功转化为SCAR标记。SCAR标记的特异性检测结果表明这两个SCAR标记只在许昌群体上有特异性扩增,可以用于许昌群体的分子检测。

Development of SCAR markers related to Heterodera filipjevi Xuchang population

Heterodera filipjevi is one of the important pathogenic nematodes that endanger the cereal crops in temperate zone, and it caused serious influence of the yield and quality. H. filipjevi is a newly identified species of cereal cyst nematode in central China. More than three years studies showed that Xuchang population is a new pathotype of H. filipjevi, and its pathogenicity was stronger than other populations of cereal cyst nematode in Huang-Huai Plain. This study aims to establish a rapid and accurate molecular detection system of Xuchang population, lay the foundation for monitoring and controlling Xuchang population of H. filipjevi and provide the basis for breeding and utilization of resistant varieties. This study focused on using RAPD-SCAR technique to analyze 9 CCN populations of 5 different CCN pathotpyes from Huang-Huai Plain. Moreover, 8 extra CCN populations were added to verify the specificity and validity of SCAR markers. 331 random primers were screened and two random primers with RAPD method were found related to Xuchang population, Primer S86, S178 could product polymorphic bands of Xuchang population about 550 bp and 1 200 bp respectively. The two RAPD markers related to Xuchang population of H. filipjevi had been transformed into SCAR markers successfully. The results of the specific detection showed that the two SCAR markers can detect the Xuchang population from other CCN populations in Huang-huai floodplain. The two SCAR markers can be used to detect and identify H. filipjevi Xuchang population.