Kaye Claudia, Milazzo Joëlle, Rozenfeld Sophie, Lebrun Marc Henri, Tharreau Didier
UMR BGPI, INRA-ENSAM-CIRAD, 34398 Montpellier Cedex 05, France.
Fungal Genet Biol. 2003 Dec;40(3):207-14. doi: 10.1016/j.fgb.2003.08.001.
Although microsatellite or simple sequence repeat (SSR) markers have several advantages, few have been developed in fungi. The goal of this study was to identify and characterize SSR-containing loci in the filamentous ascomycete Magnaporthe grisea, the causal agent of rice blast disease, and to add these markers to an integrated genetic map of this species [Theor. Appl. Genet. 95 (1997) 20]. We have constructed and screened a microsatellite-enriched small-insert genomic library as well as exploited both publicly available and one proprietary databases for identification of M. grisea SSR containing sequences. Twenty-four out of 49 primer pairs designed to amplify SSR, produced unambiguous polymorphic products in our test population of six isolates. The number of alleles at each locus ranged from two to six when assayed on 3% agarose gels. Twenty-three of the primer pairs amplified polymorphic products between Guy11 and 2539, the parents of a cross from which a genetic map for M. grisea has been established. Genetic analysis showed that all the markers segregated in the expected 1:1 ratio and map positions were determined for all 23 loci.
尽管微卫星或简单序列重复（SSR）标记具有若干优势，但在真菌中开发的此类标记却很少。本研究的目的是在稻瘟病病原菌丝状子囊菌稻瘟病菌中鉴定并表征含SSR的位点，并将这些标记添加到该物种的综合遗传图谱中[《理论与应用遗传学》95(1997)20]。我们构建并筛选了一个富含微卫星的小插入基因组文库，并利用公开可用的数据库和一个专有数据库来鉴定含稻瘟病菌SSR的序列。在设计用于扩增SSR的49对引物中，有24对在我们的6个分离株测试群体中产生了明确的多态性产物。在3%琼脂糖凝胶上检测时，每个位点的等位基因数量在2到6个之间。23对引物在Guy11和2539之间扩增出多态性产物，Guy11和2539是一个已构建稻瘟病菌遗传图谱的杂交组合的亲本。遗传分析表明，所有标记均按预期的1:1比例分离，并且确定了所有23个位点在图谱上的位置。
