不结球白菜雄性不育系细胞质类型的鉴定
毕业论文关键词：细胞质类型鉴定, 细胞质雄性不育, 不结球白菜, 特异引物
IDENTIFYING THE CYTOPLASMIC TYPE OF CMS IN NON-HEADING CHINESE CABBAGE
Abstract： In this study, we observed the floral modality characters of male sterile line of non heading Chinese Cabbage,the results show that most stamens of male sterile lines are incompletely developed. Meanwhile, we analyzed the 4 type of cytoplasmic male sterile lines of non-heading Chinese cabbage, 2016-LBC-025、2016-LBC-028-4、2016-LBC-E017-4、2016-LBC-E022-13, and the mutual maintainer, ‘Suzhouqing’, by PCR technologies based on the specific primer pairs,orf138P1/orf138P2、orf224P3/orf224P4, designed from the mitochondrial genes orf138 and orf224,which would be respectively inseparably linked with the cytoplasmic male sterile characters of Ogura CMS and the Polima CMS. The results indicated that the specific 750 bp length of fragments from the 3 male sterile lines 2016-LBC-025、2016-LBC-E017-4、2016-LBC-E022-13 were amplified by using Ogura primer pair P1/P2, and 680bp from 2016-LBC-028-4 by using Polima primer pair P1/P2.While two pairs of the primer are both unavailable for amplifying the mutual maintainer “Suzhouqing”. We can speculate that the 3 male sterile lines:2016-LBC-025、2016-LBC-E017-4 and 2016-LBC-E022-13,belong to the type of Ogura CMS,and 2016-LBC-028-4 belongs to Polma CMS. What the blastn analysis stated is that the amplified 667 bp length DNA shares 99% high homologous with that of the mitochondrial gene of Polima CMS in Brassica napus L. We can make a conjecture that the cytoplasmic male sterile of 2016-LBC-028-4 might belong to the type of Polima cytoplasmic sterile.
Key words: Cytoplasmic type identification,Cytoplasmic male sterility(CMS), Non-heading Chinese cabbage, Specific primer
目 录
摘要1
关键词1
Abstract1
Key words1
引言2
1 材料与方法2
1.1 植物材料试剂 3
1.2 花器官的形态学观察比较 3
1.3植物材料DNA制备3
1.4特异引物设计3
1.5特异引物PCR扩增3
1.6特异条带回收测序4
1.6.1特异条带回收4
1.6.1基因克隆4
1.6.1测序4
1.7序列分析4
2 结果与分析5
2.1 花器官形态学观察5
2.2 特异引物PCR扩增6
2.3 序列分析6
3 讨论 8
3.1雄性不育系花器官发育8
3.2 分子标记鉴定雄性不育系胞质不育类型8
致谢9
参考文献9
不结球白菜雄性不育系细胞质类型鉴定
不结球白菜(Brassica campestris ssp.chinensis Makino)俗称小白菜，原产于东亚,属芸薹属植物，为国内最常见的绿色蔬菜之一。不结球白菜是典型的异花授粉作物，杂种优势明显，利用杂种优势得到的杂种一代往往表现出比双亲更强大的生长能力，并改善作物的品质。有三种途径可用于杂种优势的利用，即自交不亲和系、自交系和雄性不育系[1]。但利用自交不亲和系的方法自交多代后会出现生活力衰退、亲本繁殖成本高及杂交率低等缺陷，雄性不育系则不存在这些问题，因此培育雄性不育系是更好的选育优良杂交种的方法。