YS型小麦温敏不育系育性转换基因的cDNA-AFLP分析

cDNA-AFLP Analysis of Fertility-alteration Genes in a Thermo-sensitive Male Sterile Line of YS Wheat

对YS型小麦温敏雄性不育系A3017不育与可育条件下不同发育时期的幼穗提取mRNA,进行cDNA- AFLP表达差异分析,64对引物组合获得差异片段1160个。统计分析表明,单核期表达的差异片段数最多,认为这一时期是育性转换的关键时期。对其中12个在不育或可育条件下表达的差异片段进行同收、克隆、测序,经 BLAST序列比对分析表明,可育条件下的4个EST与物质转运和能量代谢过程的铁转运蛋白1和ATP合酶α亚基,基因表达调控的反转录转座子跳跃多聚蛋白和转座子相似序列同源。不育条件下有5个EST与细胞内信号传导的ZCCT转录因子和光敏色素蛋白,基因表达调控的转座酶、染色体浓缩因子和亮氨酸富集蛋白的编码基因同源。结果表明YS型小麦温敏雄性不育系A3017的育性转换与细胞内基因表达调控、物质和能量代谢及信号传导过程有关。

mRNA was extracted from young spikes of thermo-sensitive male sterile line A3017 of YS wheat at different developmental stages ,which had differentiated under fertile conditions and infertile conditions, and then analyzed in differential expression by cDNA-AFLP to obtain 1 160 differentially-expressed fragments with 64 primer combinations. The differentially-expressed fragments that expressed themselves at the uninucleate stage were statistically found to make up the largest proportion and thus this stage was thought to be the key period to the fertility altercation. Twelve of the differentially-expressed fragments, which expressed under fertile conditions or infertile condition ,were retrieved, cloned ,sequenced and aligned by BLAST,which indicated that under fertile conditions ,there were four fragments that had the same sequence homology as iron transport protein 1 and ATP sythase alpha subunit, retrotransposon hopscotch polyprotein and transposon-like sequence in substance transport and energy metabolism and under infertile conditions,there were five fragments that had the same homology as the encoding genes of zinc-finger/CCT domain transcription factor, phytochrome P450-1ike protein, transposase, chromosome condensation factor （CCF） and leucine-rich-like protein. It was accordingly revealed that the fertility alteration of thermo-sensitive male sterile line A3017 of YS wheat had something to do with intercellular gene expression regulation,substance and energy metabolisms and signal transmission.