核质互作型水稻线粒体不育基因的RAPD分析  

Analysis of RAPD Markers About Sterility Gene in Cytoplasm-Nucleolus Interacting Type Rice Mitochondrial DNA

作　　者：裴得胜[1] 向跃武[1] 李名扬[2] 蔡平钟[1] 张志雄[1] 阎文昭[1] 侯磊[2] 裴炎[2] 

机构地区：[1]四川省农业科学院生物技术核技术研究所,成都610066 [2]西南农业大学生物技术中心,重庆400716

出　　处：《Acta Genetica Sinica》2003年第4期357-363,共7页

基　　金：8 63项目 (No.2 0 0 1AA2 410 112 )~~

摘　　要：采用RAPD对水稻Ⅱ 32A、Ⅱ 32B、Ⅱ优 94 9的mtDNA进行分析 ,得到两个特异的扩增片段OPJ18 10 0 0、OPJ18 14 0 0。序列测定两个特异片段全长分别为 10 6 8bp和 14 81bp。Southern杂交结果证明了差异片段的真实性。用OPJ18 10 0 0作探针与Ⅱ 32A、Ⅱ 32B、Ⅱ 优 94 9的mtRNA斑点杂交结果表明 ,除Ⅱ 32B外 ,都在转录水平上对水稻雄性不育有一定影响。虽然OPJ18 14 0 0对应的RNA斑点杂交呈阴性 ,但其DNA序列中存在一个七碱基 5′ TTC CCTC 3′的保守系列。据报道它是atp6基因中同源重组热点区 ,可促使线粒体基因重组形成嵌合基因 ,从而对水稻雄性不育的形成起着重要作用。经同源性比较和DNA、RNA杂交证实获得的两个片段是新的与水稻细胞质雄性不育相关的mtDNA序列。Two specific fragments OPJ18 1000 and OPJ18 1400 were obtained from mitochondrial DNAs of two rice varieties,Ⅱ 32 and Ⅱ you 949,by RAPD analysis.The sequenced lengths of the two differential fragments were 1068 bp and 1481 bp,respectively.The result of Southern hybridization demonstrated that the differential fragments were true.When hybridizing with Ⅱ 32A,Ⅱ 32B and Ⅱ you 949 by OPJ18 1000 probe,we found OPJ18 1000 which had effects on rice Ⅱ 32A and Ⅱ you 949 cytoplasmic male sterility in transcription had not the same effects on Ⅱ 32B.Though RNA dot hybridization was negative,OPJ18 1400 had a conservative domain of seven base pairs DNA sequence:5′ TTCCCTC 3′,which was homologous recombination hotspot domain in atp6 gene.It was reported that 5′ TTCCCTC 3′ induced the formation of chimeric gene in mitochondrial DNA and further caused rice male sterility.As a result,the two fragments are new mitochondrial DNA fragments relevant to rice cytoplasm male sterility by homologous analysis and DNA,RNA hybridization.

关 键 词：水稻雄性不育 mtDNA RAPD分析 SOUTHERN杂交 RNA斑点杂交 

分 类 号：Q943[生物学—植物学]