白菜小孢子发育相关基因BcMF3的分离及序列分析

Isolation and Characterization of BcMF3,a Gene Expressed Only in Maintainer Line in Chinese Cabbage-pak-choi (Brassica campestris L.ssp.chinensis Makino var.communis Tsen et Lee)

利用cDNA AFLP技术对‘矮脚黄’白菜 (BrassicacampestrisL .ssp .chinensisMakinovar.communisTsenetLeecv .Aijiaohuang)雄性不育两系不育株系与可育株系的表达差异进行分析 ,结果以A18和T16引物为引物对在可育的中蕾和大蕾中筛选出一条 4 74bp特异表达的差异条带BA18 T16 ,配合RACE方法扩增了该片段的 3′和5′末端 ,获得了该基因的cDNA全长序列BcMF3。BcMF3全长 2 0 82bp ,开放阅读框长度为 175 5bp ,编码 5 84个氨基酸。序列分析结果表明 ,BcMF3基因与油菜、大白菜和拟南芥的花药特异表达果胶甲酯酶 (PME)基因具有较高的相似性 ,由此推定 。

Expression profiling was performed using cDNA-AFLP technology on floral buds of a genic male (sterile) line (A line) and a maintainer line (B line) in Chinese cabbage pak-choi (Brassica campestris L.ssp.chinensis Makino var.communis Tsen et Lee cv.Aijiaohuang).A differentially expressed cDNA fragment,BA18-T16,was obtained via selective amplification with A18/T16 primer pair.BA18-T16 was specifically associated with the maintainer line,and was expressed only in floral buds revealed by half-quantitative RT-PCR.The 3′ end and 5′ end sequences of BA18-T16 were obtained by 3′ and 5′ RACE amplification and the full cDNA,designated as BcMF3,is 2082 bp long containing a 1 755 bp open reading frame.BcMF3 shares high homology with Bp19,a pectin methylesterase from Brassica rapa L.with 99% identity at nucleotide level and 85% identity at (amino) acid level.Suggesting that BcMF3 most likely encodes a pectin methylesterase in Chinese cabbage pak-choi.