白菜雄性不育相关新基因BcMF1的分离及特征分析

Cloning and Characterization of a New Male Sterility-Related Gene BcMF1 in Brassica campestris L. ssp. chinensis Makino

【目的】筛选与白菜雄性不育性状相关的新基因,为研究植物雄性不育的分子机制提供依据。【方法】利用cDNA-AFLP技术分析白菜核雄性不育两用系‘ZUBajh97-01AB’的表达差异,在可育株群中扩增出1条特异条带BcMF-A15T17,通过RACE和PCR技术扩增得到该基因的cDNA和DNA全长序列,并用Northern杂交验证了该基因的表达特征。【结果】该基因被命名为BcMF1,它的cDNA全长为1684bp,基因全长为1985bp,BcMF1基因仅在两用系可育株群的中、大花蕾中特异表达。推测的BcMF1蛋白含有471个氨基酸,与拟南芥未知功能蛋白家族DUF1216成员的相似性较高,预测该蛋白是一个定位在胞外的分泌蛋白。【结论】BcMF1基因是一个与白菜细胞核雄性不育相关的新基因。

[Objective] This paper aims to screen a novel gene related with the male sterility in Chinese cabbage for studying the molecular mechanism of plant male sterility. [ Method] The gene differential expression analysis was performed by cDNA-AFLP in the genic male sterile-fertile line ZUBajh97-OIAB of Chinese cabbage, and a differentially expressed cDNA fragment, BcMF-A15T17, was found in fertile line. The full cDNA and DNA sequences of the gene related with BcMF-A15T17 were amplified by RACE and PCR, and the expression characteristics of the gene were proved by Northern blot analysis. [Result] The gene, designated as BcMF1, was 1 684 bp long in cDNA and 1 985 bp long in DNA, and specially expressed in middle and big floral buds of the fertile line. The hypothetical protein BcMF 1 including 471 amino acids, which was predicted being a extracellular protein in secretory pathway, had higher similarity in the amino acids sequence to some members from a unknown protein family DUF 1216 in Arabidopsis thaliana.[ Conclusion] All the results in this study suggest that BcMF1 is a new gene related with genic male sterility in Chinese cabbage.