【作者】 符庆功；

【导师】 曹家树；

【作者基本信息】 浙江大学 ， 蔬菜学， 2003， 硕士

【摘要】 白菜(Brassica campestris L.ssp. chinensis Makino)属十字花科芸薹属芸薹种植物，在我国农业生产和人民生活中占有非常重要的地位。白菜杂种优势十分明显，其雄性不育系利用一直被认为是目前一代杂种生产的理想途径，因而，有关其雄性不育发生的机理研究一直深受人们的重视。分子生物学技术的发展则为从分子水平阐明白菜雄性不育发生机理提供了重要的工具。 本研究以‘矮脚黄’白菜核不育两用系的测交后代群体为实验材料，利用AFLP指纹图谱技术研究了可育系与不育系基因组DNA多态性差异，并对差异条带进行了克隆、测序。探讨了所得差异片段与先前所得的育性相关基因之间的关系。结果如下： (1)本实验分别以可育系和不育系基因组DNA混合池为模板，选用64对E+3／M3引物组合进行选扩筛选，分析得到了两条可育系所特有的差异条带MF-14和MF-24。 (2)以回收得到特异片段MF-14为探针，通过Southern杂交检测到白菜不育系中也含有该片段的同源序列。并初步探讨不育系碱基序列突变情况。 (3)序列测定表明特异片段MF-14长289bp；比较特异片段MF-14与先前所得育性相关片段S107及育性相关基因CYP86MF、BCMF1、BCMF2、BCMF3、BCMF4之间序列关系，结果显示它们之间的相似性均小于50％。分析结果表明MF-14是可育系中发现的一个新的特异片段。 (4)以MF-14特异片段为基础，设计引物进行了SCAR标记转换研究。更多还原

【Abstract】 Chinese cabbage-pak-choi (Brassica campestris L. ssp. chinensis Makino) is a typical allogamy plant and is of attractive heterosis. And, it is also one of the most popular leafy vegetables, especially in the South China. Much attention, for which, has been paid to the research of male sterility mechanism and its application in it inthe past several decades. Now, with the help of progressing molecular biotechnology, it has become possible of explaining the molecular mechanism of male sterility.’Aijiaohuang’ used in this study is a representative genie male sterile AB lines in Chinese cabbage-pak-choi, and it is also an optimal plant material in the research of male sterility mechanism. In order to compare the differences of genomic DNA between the fertile line and the sterile line with AFLP technique, genomic DNA bulks of the fertile and the sterile lines of which were built up. By using different primer combinations, some polymorphism bands were determined. Meanwhile, the cloning and sequencing of which was also performed .and the main results were as follows:(1) Only two obvious polymorphism bands were obtained in the fertile line from selective PCR amplification with 64 pairs E+3/M+ rimer combinations, and which were named as MF-14 and MF-24, respectively. Moreover the polymorphism band MF-14 was cloned successfully.(2) It proved that there should be nucleotides sequence similar to MF-14 in the sterile line by the Southern hybridization using the fragment MF-14 as probe. Basing that, the mutation of nucleotides sequence in the region related to fertile gene was discussed.(3) Nucleotides sequencing revealed that the fragment MF-14 consists of 289 base pairs. Comparison of sequences similarity between MF-14 and other specific fragment or genes (S107, CYP86MF, BCMF1, BCMF2, BCMF3, BCMF4), which got in the previous research of male sterile, show that all the similarity between them were below 50%. So MF-14 was believed to be a newly specific DNA fragment, according to which, in the fertile line in ’Aijiaohuang’.(4) The sequence characterized amplified region marker about the specific fragment MF-14 was discussed.更多还原