多年生簇毛麦基因组长末端重复序列片段的克隆、定位与应用  

Isolation,Mapping and Application of An LTR Fragment for Dasypyrum Genome

作　　者：刘成[1] 杨足君[1] 李光蓉[1] 冯娟[1] 周建平[1] 任正隆[1] 

机构地区：[1]电子科技大学生命科学与技术学院,四川成都610054

出　　处：《作物学报》2006年第11期1656-1662,共7页Acta Agronomica Sinica

基　　金：国家自然科学基金(30170502和30170579)

摘　　要：以多年生簇毛麦、普通小麦中国春、山羊草等为材料,用142个10碱基随机引物进行RAPD分析,筛选出1个簇毛麦的特异性RAPD片段,经克隆测序得其全长为1 182 bp(登录号DQ167398),被命名为OPH121182。序列比对结果显示,OPH121182与栽培大麦中的反转重复序列Sabrina有95%的同源性,表明OPH121182也是一类反转重复序列。原位杂交结果显示,pDbH12在多年生簇毛麦所有染色体上除端部和着丝点以外的所有区域均具有很强的杂交信号,说明OPH121182是簇毛麦基因组的一个特异高度重复序列。进一步以OPH121182的DNA序列为基础设计特异PCR引物,对供试材料进行扩增,表明OPH121182分布在簇毛麦全部染色体上,可作为分子标记检测具有簇毛麦染色质的后代和小麦-簇毛麦双二倍体。Dasypyrum genus is an important source for improving the genetic variability of wheat （ Triticum aestiruum ）. However, the tetraploid species, D. breviaristatum, has not been reported to introduce into wheat. In this study, random amplified polymorphie DNA （RAPD） analysis was performed on Dasypyrum breviaristatum and other 9 Triticeae materials by 142 random 10-based primers. A 1 182 bp Dasypyrum specific RAPD fragment was obtained, which was named OPH 12 1182 .Sequence similarity analysis revealed that 95 % of OPH12 1182 was similar to a retrotransposon repeat sequence Sabrina of Hordeum vulgate, indicating that OPH12 1182 was also a kind of retrotransposon repeat sequence. The result of fluorescenee in situ hybridization （FISH） suggested that pDbHl2 was specifically hybridized throughout all D. breviaristatum chromosomes arms exeept for the terminal and centromeric regions, which indicated that OPH12 1182 is a specific repetitive sequence in genome of Dasypyrum. Furthermore, based on the sequence of OPM12 1182, pairs of primers were designed, and used to amplify materials used in this study, the result showed that all the seven pairs of Das）pyrum chromosomes contained the specific DNA segment and OPH12 1182 could be used to detect offsprings containing Dasypyrum ehromatins and wheat-Dasypyrum amphiploids. Therefore, OPM12 1182 could be used as a molecular marker for detection of chromatins of Dasypyrum.

关 键 词：簇毛麦 重复序列 克隆 原他杂交 特异PCR 

分 类 号：S512.1[农业科学—作物学]