摘要
以4个限制性内切酶(EcoRI、BamHI、HindIII、PstI)进行酶切、14个线粒体基因为探针,对NCa和pol、nap、ogura、tour等5个甘蓝型油菜细胞质雄性不育系的不育胞质线粒体DNA进行RFLP分析。在总计54个有效探针/酶组合中,除12个组合在5种不育细胞质间没有检测到差异外,其余42个探针/酶组合在5个不育系中均检测到差异。其中,atp1/EcoRI、atp1/HindIII、atp6/EcoRI、atp8/BamHI、orf139/BamHI等5个探针/酶组合在5个不育系的不育胞质间检测到各不相同的条带,从而可以将它们两两完全区分开。RFLP分析结果从分子水平上进一步证明了NCa不育系确实具有与pol、nap、ogura、tour等不育系不同的的不育胞质,是一类新的不育胞质类型。这5个探针/酶组合也可以作为鉴定甘蓝型油菜不育细胞质的分子标记。根据控制pol、nap、ogura等不育系的线粒体基因序列和拟南芥线粒体基因组序列设计了22对引物,对上述5种甘蓝型油菜不育系的线粒体DNA进行了PCR分析,没有找到可以将NCa不育胞质与其它类型不育胞质明显区分开来的引物。
Previous research on the NCa, a cytoplasmic male sterile （CMS） line in Brassica napus, showed that it might hold a new type of sterile cytoplasm. RFLP analysis of the mitochondrial （mt） DNA of the 5 CMS lines, i.e., NCa, pol, nap, ogura and tour CMS, combining 14 probes originated from mt genes with 4 endonuclease, such as EcoR Ⅰ, BamH Ⅰ, Hind Ⅲ and Pst Ⅰ, was performed and detected that 42 combinations of probe/enzyme out of the total 54 combinations could discover the different bands among the 5 CMS lines and then distinguish some of them from the others. Moreover, 5 combinations of atpl/EcoR Ⅰ, atpl/Hind Ⅲ, atp6/EcoR Ⅰ, atp8/BamH Ⅰ, and orf139/BamH Ⅰ, could powerfully distinguish the 5 CMS lines from each other. These 5 combinations of probe/enzyme could be exploited as markers to identify the cytoplasmic type of rapeseed CMS. These results definitely demonstrated the NCa CMS hold a new type of sterile cytoplasm different from pol, nap, ogura and tour CMS. Twenty-two primers were designed according to the Arabidopsis mt genome and the mt regions which controlled the cytoplasmic male sterility of pol, nap, ogura CMS lines, and were used in PCR analysis of mt DNA from the above five CMS lines. The PCR results proved that no one of the 22 primers could distinguish the NCa CMS from the other four.
出处
《中国农业科学》
CAS
CSCD
北大核心
2005年第10期1965-1972,共8页
Scientia Agricultura Sinica
基金
国家"863"计划资助项目(2002AA207009)
武汉市青年晨光计划项目(20035002016-36)
农业部油料作物遗传改良重点开放实验室开放课题
