细胞质雄性不育高粱叶绿体ndh D基因的序列变异(英文)  

Sequence Variation of the Chloroplast Gene ndh D Region in Cytoplasmic Male Sterile Sorghum

作　　者：范昌发[1] 孙春昀[1] 郭骁才[1] 张福耀[2] 孙毅[3] 牛天堂[3] 贾敬芬[4] 

机构地区：[1]中国科学院成都生物研究所,成都610041 [2]山西省农业科学院高粱研究所,榆次030600 [3]山西省农业科学院农业生物技术中心,太原030031 [4]西北大学生命科学学院,西安710069

出　　处：《Acta Genetica Sinica》2002年第10期907-914,共8页

基　　金：中国科学院知识创新工程项目;中科院成都地奥科学基金资助

摘　　要：片段SAAU - 0 2 70 0 特异地扩增自 7种具可育细胞质的高粱材料的总DNA ,含有叶绿体psaC(88bp)和ndhD(192bp)基因的部分序列。该片段与EcoRⅠ +HindⅢ酶切的总DNA、线粒体DNA和叶绿体DNA杂交 ,在总DNA中获得了 0 .74kb的杂交带 ,而在叶绿体中获得 0 .74kb和 0 .4 5kb两条杂交带 ,与线粒体DNA无杂交 ;与经HaeⅢ酶切的总DNA杂交 ,在不育系中获得 4 .9kb的杂交带 ,而保持系的杂交带为 4 .4 5kb。参考GenBank中高粱的近缘物种玉米叶绿体基因组的序列 ,构建了ndhD基因区的酶切位点图谱 ,借此分析得出高粱不育系的叶绿体ndhD基因序列已发生改变。这种变异与高粱细胞质雄性不育发生的关系正在探讨中。A fragment SAAU-02 700 was amplified specifically from total DNA of seven sorghum varieties with male fertile cytoplasm (N cytoplasm). PCR assays indicated that it was amplified from chloroplast (cp) DNA. Sequence analysis revealed this newly cloned fragment contained a portion of chloroplast gene psa C (88bp) and part of ndh D gene (192bp). Total DNA,mitochondrial (mt) DNA,and cpDNA were digested with Eco R Ⅰ+ Hin dⅢ and probed with fragment SAAU-02 700 . The Southern hybridization patterns displayed a 0.74kb band both in total DNA and cpDNA,but an additional faint band 0.45kb in size was found only in the latter. No polymorphic hybridization signal between the N-cytoplasm and male sterile cytoplasm (S cytoplasm) was observed. Southern hybridization of total DNA of CMS line A 1Tx623 and fertile line Tx623 digested with Hae Ⅲ gave a band 4.9kb in size in the former and a 4.45kb band in the latter. This revealed that the sequence of ndh D from CMS line was likely altered. Further studies designed to determine whether or not the variation has some effect on the metabolism of mitochondria and chloroplast,even on the occurrence of male sterility in sorghum are underway.

关 键 词：细胞质雄性不育 高粱 叶绿体 ndh D基因 序列变异 

分 类 号：Q943[生物学—植物学]