小麦K型不育系育性恢复基因的cDNA-AFLP分析

Identification of restorer genes of K-type CMS in wheat by cDNA-AFLP

采用cDNA-AFLP技术,对小麦K型不育系豫麦3号及其相应的保持系,以及2个利用恢复系豫麦2号构建的不育系豫麦3号遗传背景的恢复基因近等基因系在小麦二核花粉时期的基因表达谱进行了分析.共筛选120对引物,产生10 757个转录本(TDF),用26对引物检测到53个(4.9%)差异TDF.经大规模克隆、测序分析,最终获得35个差异TDF,并对这些差异TDF进行了电子延伸.经Blastx比对和功能分类分析,其中23条TDF与NCBI有序列同源性,功能主要涉及细胞和组织代谢(37.1%)、转录因子(8.6%)、胁迫相应(5.7%)、转录促进(5.7%)、表达调节(5.7%)以及混合功能(29%)等;3条TDF(8.6%)与未知功能蛋白同源性较高;其余9条TDF(25.7%)未找到同源性匹配,可能是一些新基因.

cDNA-amplified fragment length polymorphism （cDNA-AFLP） analysis was employed to identify the different expression of restorer genes between K-type sterile line Yumai 3 and NILs of re- storer genes with the background of Yumai 3 during their dicaryophase of anther development. The re- suits showed that over 10 757 expression bands existed between the two materials. Among about 53 transcript-derived fragments （TDFs） , 35 TDFs were cloned, and all of them were electronically ex- tended. According to the genomic database, 23 TDFs identified were obviously homologous to the genes with the known or putative function, 3 TDFs were homologous to uncharacterized genes while 9 TDFs did not show significant matches to any genes in the Genedatabase.