中国春Ph1基因的RAPD标记鉴定  

Identification of RPAD Markers for Ph1 Gene in Common wheat Chinese Spring

作　　者：王新望[1] 赖菁茹[2] 姚大年 孙辉[1] 刘广田[1] 

机构地区：[1]中国农业大学植物遗传育种系,北京100094 [2]河南农业科学院小麦所,郑州450002

出　　处：《农业生物技术学报》1999年第1期29-35,共7页Journal of Agricultural Biotechnology

摘　　要：用260个10bp随机引物对中国春（CS）、ph1b突变体及其F2分离群体基因组DNA进行RAPD分析，筛选出两个特异的RAPD标记OPR7936和OPR17524。经Southern分析，并结合减数分裂MI染色体配对分析，认为这两个RAPD标记位于5BL染色体ph1b基因缺失区中，可以作为ph1b基因的分子标记。经过连锁分析，测得OPR7936与Ph1基因之间的距离为11．4cM，OPR17524与Ph1基因之间的距离为5．0cM，均位于Ph1基因的远着丝粒端。测序后，经同源性比较发现这两个DNA序列可能是Ph1b基因区中的两个不同位点上的新序列，并有可能成为Ph1基因区图谱新的探针。根据这两个PAPD克隆片段的两端序列，分别设计出一对24bp的SCAR引物，可特异扩增Ph1基因的一条带（SCR7823和SCR17403），而ph1b基因缺少各自的特征带，其扩增产物无需电泳分离，只需在反应管中加入溴化乙锭，紫外灯下直接根据荧光反应即可鉴定出ph1b基因型，为今后ph1b基因的转育进行标记辅助选择又提供了两个易于操作、有效的分子标记。Two RAPD markers, OPR7936 and OPR17524, which closely linked to Ph1 gene in Chinese Spring wheat, were identified by using 260 10-mer random primers for the total DNAs of Chinese Spring (CS), ph1b mutant and the F2 population. OPR7936 and OPR17524 were located on the interstitial deletion region of Ph1 gene and distal to Ph1 gene apart from 11.4 cM and 5.0 cM respectively. Two SCAR markers were developed based on the sequences of these tWo RAPD clones. The conditions were optimal to produce only one band for CS, none for ph1b mutant. So the products of SCAR analyses could be detected directly by addition of ethdium bromide under UV light in order to screen ph I b homozygotes. This strategy is less time-consuming and convenient for the marker-assisted selection of Phlgene in the wheat breeding program.

关 键 词：中国 春小麦 Ph1基因 Ph1b突变体 RAPD 

分 类 号：S512.103.2[农业科学—作物学]