扁蓿豆冷诱导基因差异表达分析.pdfVIP

西北植物学报， 2013 ， 33 (1) : 0060一 0065 Acta Bot. Boreal. -Occident. Sin. 文章编号 :1000-4025(2013)01-0060-06 扁蓓豆冷诱导基因差异表达分析 张宇1 ，石凤刽l\云娜1 ，高 霞飞伊风艳l ，蔡丽艳2 (1内蒙古农业大学生态环境学院，呼和浩特 010019;2 中国农业科学院草原研究所，呼和浩特 010010) 飞揍F 摘 要 2 以直立型扁稽豆幼苗为试验材料，采用 cDNA-AFLP 技术分析扁稽豆在低温胁迫诱导下的基因表达差异。 结果显示，利用筛选的 64 对引物组合，对 o .C低温处理 3~5 叶期扁稽豆幼苗的叶片 cDNA 进行扩增，共获得 549 条差异表达的转录衍生片段CTDFs) 。选取上调表达较好的 43 条片段进行克隆、测序、 Blastx 比对和功能分析，其 中 32 个 TDFs 的蛋白序列与基础代谢、信号转导、转录因子、防御等功能有关， 11 个 TDFs 为假设蛋白、未知蛋白 或没有找到一致序列。利用荧光定量 PCR 对 3 种不同上调表达差异片段进行验证，结果可从数值上更准确地显 示差异片段在低温胁迫过程中的相对表达量。 关键词:扁稽豆;冷诱导 ;cDNA-AFLP;差异表达 中图分类号 :Q786 文献标志码 :A cDNA-AFLP Analysis of Cold-induced Genes in Medicago ruthenica ZHANG Yu 1 ,SHI Fengling 1 费 ， YUN Na1 ,GAO Xia1 , YI Fengyan1 , CAI Liyan2 (1 College of Ecology and Environmental Science. Inner Mongolia Agricultural University. Hohhot 010019. China; 2 Grassland Re search Institute of Chinese Academy of Agricultural Sciences. 日ohhot 010010.China) Abstract: cDNA-AFLP approach was employed to analyze genes differential expression under cold stress u- sing the three-five leaf stage leaves of Medicago ruthenica. The results showed that 549 differentially ex- pressed transcript-derived fragments(TDFs) were identified by sixty-four primer combinations ,and 43 up- regulated TDFs of them were sequenced and analyzed. Blastx-analysis and functional analysis indicated that 32 TDFs might participate in senescence and metabolism pathways , signal transduction , transcription regu lation , stress responses etc , other 11 TDFs were showed homology to unknown and hypothetical function ge口es. The qRT-PCR results showed that it was accuately on the numerical value for expression of three different up-regulated TDFs were induced by low temperature stress. Key words: Medicago ruthenica ; cold-induced; cDN A-AFLP; different expression 直立型扁稽豆 [Medicago ruthenica (L.) cv. ZhilixingJ是从野生扁稽豆选育出的高产、高抗牧草 新品种，可在首稽不易生长的干旱、寒冷地区种植， 40 oc 环境条件下也能安全越冬[叫，可作为改良 同属牧草抗逆性的优质基因资源[4-5J 。因此，分离和 筛选扁稽豆抗寒相关表达基因，研究其抗寒分子机 收稿日期: 2012-