小麦中国春背景下长穗偃麦草Ee染色体组特异AFLP及STS标记的建立

为建立长穗偃麦草Ee染色体组特异的分子标记，以普通小麦中国春、中国春-长穗偃麦草二体代换系、附加系为材料，用5对AFLP引物进行分析，共筛选出28个分布于1Ee-7Ee 7个染色体特异的AFLP标记，经PAGE凝胶电泳后回收、克隆和测序，并根据测序结果设计PCR特异引物，成功地将AFLP标记E-ATC/M-CGTA341bp, E-ATT/M-CTAG265bp, E-AAT/M-CCAG139bp和E-ATT/M-CTAG205bp转化为实验结果稳定、操作更简单的STS标记。利用获得的STS标记对5个长穗偃麦草居群和8个小麦品种进行了鉴定。结果表明其可以在长穗偃麦草居群中被检测到，但在其它小麦背景中检测不到。表明这些标记可以用于小麦-长穗偃麦草异源附加系和代换系中长穗偃麦草遗传物种的检测。

Development of Ee-chromosome Specific AFLP and STS Molecular Marker for Lophopyrum elongatum in Chinese Spring Wheat Background

For development of the Ee chromosome specific molecular markers for diploid species of L.elongatum, amplified fragment length polymorphism (AFLP) analysis was performed on common wheat Chinese Spring (CS), CS—L. elongatum disomic substitution and additional lines using 5 primer combinations, and 28 AFLP markers, which were specific for 1Ee to 7Ee chromosome of the diploid species of L.elongatum, respectively, were obtained. After cloning and sequencing of these AFLP fragments, four AFLP markers of E-ATC/M-CGTA341bp, E-ATT/M-CTAG265bp, E-AAT/M-CCAG139bp and E-ATT/M-CTAG205bp were successfully converted into reliable sequence tagged markers (STS). These four markers were further subject to amplification other five accessions of the L.elongatum and other eight common wheats without L.elongatum sources for validating their specificity and polymorphism. The results showed that these markers could amplification the same size of the fragment from L.elongatum accessions but couldn’t amplification it from other eight common wheat, suggesting that these markers could be used for identify and tracing the Ee chromosomes in wheat- L.elongatum substitution and additional lines.