大白菜雄性核不育基因的染色体定位及AFLP分子标记筛选  

Chromosomal Location and AFLP Marker Screening of Genic Male Sterile Gene in Chinese Cabbage

作　　者：袁鹤[1] 张成合[1] 刘海河[1] 轩淑欣[1] 李晓峰[1] 申书兴[1] 

机构地区：[1]河北农业大学园艺学院,河北保定071001

出　　处：《中国农业科学》2009年第6期2061-2067,共7页Scientia Agricultura Sinica

基　　金：河北省自然科学基金项目(C2006000450)

摘　　要：【目的】基因定位是遗传研究的重点内容,也是育种工作的重要基础。本研究以大白菜雄性不育两用系‘大阳AB系’和大白菜系列初级三体为材料,对决定大白菜雄性不育性的核基因(ms)进行染色体定位和分子标记筛选,旨在为其分子克隆和遗传利用奠定基础。【方法】基因的染色体定位采用初级三体遗传分析和χ2测验法,分子标记采用AFLP分析方法。【结果】大白菜‘大阳AB系’的雄性不育系(msms)×大白菜系列初级三体(Tri-1、Tri-2、Tri-3、Tri-4、Tri-5、Tri-6、Tri-7、Tri-8、Tri-9)的9个杂交组合中,F1均为可育株;从F1选出三体植株进行自交和测交,其自交子代(F2)和测交子代(Tc)中可育株与不育株的分离比例,只有Tri-4表现为15.5﹕1(F2)和3.24﹕1(Tc)、不符合孟德尔的单基因遗传分离比例(3﹕1和1﹕1),其它三体在2.60﹕1～3.76﹕1(F2)和0.81﹕1～1.48﹕1(Tc)、均符合孟德尔的单基因遗传分离比例(3﹕1和1﹕1),这表明该雄性不育性的遗传与Tri-4的遗传表现一致,即确定该雄性不育性的基因位于大白菜4号染色体上。基于染色体和染色单体分离的三体遗传分析表明,该雄性不育基因位于4号染色体的近着丝点区域,通过对124个AFLP引物组合的筛选,初步获得了一个与雄性核不育基因位点连锁的AFLP标记E34M49(398bp),遗传距离约为1.2cM。【结论】利用初级三体遗传分析首次将大白菜雄性核不育基因定位在第4号染色体上。[ Objective ] Chromosome localization of genes is an important work for genetic study and breeding practice. In this paper Dayang AB line and trisomics of Chinese cabbage were used for assigning the male sterile gene to a particular chromosome and screening the molecular markers linked to the locus. [ Method] Locating the male sterile gene to a particular chromosome was conducted by primary trisomic analysis and X^2 test, and screening the molecular markers linked to the gene locus by AFLP techniques [Result] All FI progenies in the nine combinations derived from sterile line×Tri-1 to sterile line x Tri-9 were male ferterile. This indicated that the male fertility was dominant to the sterility in Chinese cabbage Dayang AB line. Selecting the trisomic（2n＋1） plants from each FI progenies and being selfed or test-crossed to the sterile line, and the results showed that only Tri-4 combination displayed 15.5 ： 1 segregation proportion of male fertile plants to male sterile plants in F2 and 3.24 ： 1 in Tc （test-cross）, which disobeyed Mendelian segregation ratios for a monogenetic factor, while other combinations varied from 2.60：1 to 3.76 ： 1 in F2 and 0.81 ： 1 to 1.48 ： 1 in Tc, which fitted Mendelian segregation proportions （3 ： 1 and 1 ： 1） for a monogenetic factor. These results indicated that the gene controlling the male sterility of Chinese cabbage Dayang AB line was associated with trisomic 4, i.e. the gene was located on the chromosome 4. Further trisornic analysis based on the chromosomal segregation （assuming the gene closely linked to the centromere ） and on the chromatid segregation（assuming the gene apart from the centromere ） showed that the gene was closer to the centromere. Furthermore, an AFLP marker （E34M49） linked to the sterile gene was preliminarily screened out from 124 random primers. [Conclusion] Using primary trisomic analysis and molecular screening techniques, the sterile gene of Chinese cabbage was assigned to chromosome 4 and an AFLP marker （3

关 键 词：大白菜 初级三体 雄性不育 基因定位 AFLP标记 

分 类 号：S634.1[农业科学—蔬菜学]