白菜核雄性不育两用系AFLP扩增特异片段的克隆及序列分析

Cloning and sequence analysis of a specific amplified fragment related to genic male sterility AB lines of Chinese cabbage (Brassica campestris L. ssp. chinensis Makino)

利用AFLP技术从白菜核不育两用系"矮脚黄"可育系DNA中得到一个特异扩增片段MF-14.Southern杂交结果证实不育系也存在该片段的同源序列;回收该特异扩增片段并将其克隆到pGEM-TEasy载体进行序列测定.测序结果表明,该片段全长289bp,其碱基组成为A+T=46.02%.与白菜RAPD特异片段S107及其他育性相关基因核苷酸序列比较,同源性均低于50%.表明该片段为一新的序列.

A specific amplified DNA fragment, MF-14, was obtained from fertile lines of Chinese cabbage pak-choi (Brassica campestris L.ssp. chinensis Makino var. communis Tsen et Lee cv. Aijiaohuang) by AFLP technique. It was proved by Southern hybridization analysis that the similar nucleotide sequence was present in the sterile line. This specific fragment was cloned into pGEM-T Easy vector and DNA sequence was determined. The result showed that it consist of 289 bp, of which the base components of A+T accounted for 46.02%. Comparison between MF-14 and other specific sequences derived from Chinese cabbage GMS AB lines revealed that the homology in most cases was below 50%. This implied that the above mentioned fragment represented a newly found DNA sequence in Chinese cabbage.