吴博—毕业论文.docVIP

小麦K型不育系育性恢复基因的cDNA-AFLP分析 吴博 （农学院农学专升本2012-2班） 摘要：采用优化的cDNA-AFLP体系，对小麦K型不育系豫麦3号及其相应的保持系以及两个恢复基因近等基因系BC9F2在小麦二核花粉时期的基因表达谱进行了分析。通过120对引物筛选，产生10757个转录本(TDF)，用26对引物检测到53个差异TDF。经大规模克隆、测序分析，最终获得35个差异TDF。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、转录因子、胁迫相应、转录促进、表达调节以及混合功能等；3条TDF与未知功能蛋白同源性较高；其余9条TDF未找到同源性匹配，可能是一些新基因。通过对功能已知的基因分析，推测吲哚乙酸合成酶与脯氨酰胺合成酶可能与花药育性恢复的过程相关。 关键词：小麦；K型不育系；育性恢复基因；cDNA-AFLP；差异表达基因Identification of restorer genes of K-type CMS in wheat by cDNA-AFLP Wu-Bo (Agriculture 2012-2 of Agronomy College） ABSTRACT: Optimized cDNA-AFLP analysis was employed to identify gene expression profile of K-type sterile line Yumai 3 and its maintainer line and two NILs of restorer genes BC9F2 during their dicaryophase of anther development. After the 120 pairs of primer screened, the results showed that 10 757 transcript-derived fragments (TDFs) existed between the materials, and 53(4.9%) discrepant TDFs was detected by 26 pairs of primer. After large scale cloning and sequencing, eventually 35 different TDFs obtained, and all of them were extended by electronic method. By means of Blastx comparison and functional classification analysis, 23 TDFs of them have sequence homology with NCBI, and their function mainly relates to cell and tissue metabolism(37.1%), transcription factor(8.6%), Stress corresponding(5.7%), transcription promoting(5.7%), regulation of gene expression(5.7%) and mixed function (29%); 3(8.6%)TDFs are highly homology with unknown function protein while 9 TDFs(25.7%) do not show significant matches to any genes in the Genedatabase. they may be new genes. By analyzing the function of known genes, presumably, indole acetic acid synthetase and preserved ammonia amide synthetase may be associated with the process of anther fertility restoration. Key words: wheat; sterile line with Ae.kotschi; restorer gene; near-isogenic line; cDNA-AFLP; differential expression 小麦是我国第三大粮食作物，也是世界上种植范围最广的粮食作物。随着全球人口剧增，人类对小麦的需求量也呈增长趋势。因此，培育高产小麦新品种已经成为当前小麦育种的迫切要求[13]。杂种优势的利用能大