苜蓿雄性不育系MS-4SSH文库构建及基因表达分析

摘要

The buds of male sterile line of alfalfa MS-4 and it's maintainer were taken as materials,employed the suppression subtractive hybridization (SSH) technology.Two differential gene expression subtractive cDNA libraries were constructed under sterile and fertile conditions.Positive clones were randomly selected to sequence and analyze by Blast.190 ESTs with known functions were got.6 genes were inferred which may be associated with male fertility of alfalfa by GO functional classification.The expression of 6 genes were analyzed by fluorescence quantitative PCR,the results showed 6 genes had differential expression at different development stages in the male sterile line of alfalfa.It suggested that these 6 genes may be related to the male fertility of alfalfa,and play an important role in fertility transformation.%以苜蓿雄性不育系MS-4与可育系花蕾为材料,利用抑制性消减杂交(SSH)技术,分别构建了不育条件下和可育条件下基因差异表达消减cDNA文库,在2个库中分别随机挑选阳性克隆进行测序,经序列比对分析,共获得功能已知的EST序列190条.后经GO功能分类,对推定出的6个相关基因进行荧光定量PCR分析.结果表明:6个基因在苜蓿雄性不育系花蕾不同发育时期中均出现差异表达,推测这6个基因可能与苜蓿雄性育性相关,并且在其育性转换中具有重要作用.