基于EST-PCR的簇毛麦染色体特异分子标记筛选及应用

为定位、转移和利用簇毛麦有益基因, 通过花粉辐射, 获得一批包括小麦-簇毛麦易位染色体的异染色体系。为了鉴定这批材料中的簇毛麦染色体身份, 根据水稻、小麦的EST序列合成了240对STS引物, 其中34对引物在普通小麦中国春与簇毛麦间存在多态性；进一步对亲本及簇毛麦二体异附加系进行PCR扩增分析, 标记CINAU32_-300可追踪簇毛麦1V染色体, 标记CINAU33_-280、CINAU34_-510、CINAU35_-1100、CINAU36_-380和CINAU37_-400可追踪簇毛麦2V染色体, 标记CINAU38_-250可追踪簇毛麦3V染色体, 标记CINAU39_-950和CINAU40_-800可追踪簇毛麦4V染色体, 标记CINAU41_-745和CINAU42_-1050可追踪簇毛麦5V染色体, 标记CINAU44_-765和CINAU45_-495可追踪簇毛麦7V染色体。加上本室已开发的2个6V染色体特异标记, 用这些簇毛麦特异分子标记鉴定辐射诱导材料的部分回交后代, 选育出小麦背景中只包含单条簇毛麦染色体的整套1V至7V染色体系, 同时有18条易位染色体的簇毛麦身份得到确定, 表明这些标记可以用来快速检测普通小麦背景中的簇毛麦染色体或染色体片段。

Screening and Application of EST-based PCR Markers Specific to Indi-vidual Chromosomes of Haynaldia villosa

To localize, transfer and use interested genes of Haynaldia villosa, a series of wheat-H. villosa alien chromosome lines including translocation lines induced by irradiation have been developed. To identify H. villosa chromosomes or chromosome segments in these materials, 240 STS primer pairs were designed based on EST sequences of rice and wheat, and 34 of them could amplify specific polymorphic bands between common wheat cv. Chinese Spring and H. villosa. These 34 STS primer pairs were further used for screening markers specific to individual chromosomes of H. villosa using 1V to 7V disomic additional lines and their parents. The results showed that the marker CINAU32_-300 could be used to trace chromosome 1V, CINAU33_-280, CINAU34_-510, CINAU35_-1100, CINAU36_-380 and CINAU37_-400 to 2V, CINAU38_-250 to 3V, CINAU39_-950 and CINAU40_-800 to 4V, CINAU41_-745 and CINAU42_-1050 to 5V, CINAU44_-765 and CINAU45_-495 to 7V. These EST-STS markers combined with the two 6V specific markers previously in our institute were used to identify the H. villosa chromosome and chromosome segments of backcrossed generations derived from pollen irradiation. A set of 1V to 7V addition lines and 18 translocation chromosomes involved in different H. villosa chromosomes were identified. Therefore, these chromosome-specific EST-STS markers could be used to detect chromosomes and chromosome segments of H. villosa in common wheat background.