与大白菜细胞质不育相关RAPD特征片段的克隆和序列分析

Cloning and Sequencing for RAPD Specific Fragments in Chinese Cabbage Cytoplasmic Male Sterile

为了研究和获得大白菜细胞质不育系和保持系线粒体DNA的多态性,选用153个RAPD随机引物,对3套材料,每套中包含不同来源、同核异质的大白菜细胞质不育系(Pol-CMS、Ogu-CMS和CMS96)和保持系线粒体DNA进行RAPD分析。结果表明,有4个RAPD随机引物可以在全部Ogu-CMS和CMS96不育系中扩增出特异带,有1个随机引物仅在所有Ogu-CMS不育系中扩增出特异带;同时将OPAH16随机引物扩增的与不育相关的特异带进行了克隆和测序。结果表明,所获得的特异片段均与拟南芥和甘蓝型油菜的线粒体基因组序列部分同源,但与已发表的不育基因序列没有同源性。由这5个序列推导出的蛋白质与拟南芥中的一种未知功能蛋白AtMg00760(ORF109b)具有部分同源性。这5个序列的可能功能正在验证中。

To abtain the differences between maintainer and cytoplasmic male sterile line in Chinese cabbage（Brassica campestris L.ssp. pekinensis）, 153 random primers were used for RAPD analysis in maintainer and cytoplasmic male sterile mtDNA which had the same nucleus and different cytoplasm. Specific fragments could be amplified by 4 RAPD primers in all Ogu-CMS and CMS96. And specific fragments related to all Ogu-CMS could be amplified by one RAPD primer. 5 specific fragments amplified by OPAH16 primers in all Ogu-CMS and CMS96 were cloned. Then sequences were compared by Blastn method. Results show that these sequences are homological to Brassica napus and Arabidopsis mtDNA genome partially. But they are not homological to CMS gene reported before. The 5 sequences can be induced a protein that is homological to ORF109b in Arabidopsis mtDNA genome. The functions of the 5 sequences are testing.