柠条锦鸡儿CBF%2fDREB1基因克隆及其过表达对提高拟南芥抗旱、抗冻能力影响.pdf

摘 要 低温、干旱和盐害等非生物胁迫严重影响植物的生长、发育和作物产量。DREB 转录因子，能够特异性地结合DRE 元件上的核心序列A/GCCGAC，启动下游与抗逆 相关的功能基因的表达来响应对低温、高盐、干旱等胁迫。基于 DREB 在信号途径 中的重要地位，本论文以柠条锦鸡儿 DREB1 基因为研究对象，以拟南芥野生型、 CkDREB1 基因过表达转基因纯合体植物为材料，进行了初步分析，结果如下： （1）从柠条锦鸡儿中成功分离了三个版本的CkDREB1 基因的全长DNA 序列，开放 阅读框全长均为615bp 。CkDREB1 蛋白AP2 上游和下游有 DREB1 亚家族成员一般 都有的PKK/RPAGRxKFxETRHP 保守序列和DSAWR 保守序列，序列的同源性比对 分析结果表明CkDREB1 蛋白属于DREB 蛋白家族的A-1 亚家族。 （2 ）Northern 分析表明CkDREB1 基因受低温强烈诱导，对干旱和ABA 也有一定的 响应，但对高盐处理不响应。 （3 ）RT-PCR 和Northern 检测表明CkDREB1 基因过表达纯合体株系中CkDREB1 均 有不同程度的过表达。冷驯化后，CkDREB1 基因过表达株系的抗冻能力比野生型提 高；干旱胁迫下，CkDREB1 基因过表达株系叶片的持水能力比野生型更强。 关键词：柠条锦鸡儿；DREB1 ；过表达；拟南芥；抗冻；抗旱 Cloning of CBF/DREB1 of Caragana korshinkii Kom and the Effect of Its Overexpression on Drought and Cold Resistance of Transgenic Arabidopsis thaliana Abstract Low temperature, drought and salt stresses are disadvantageous for the plants growth and development. Transcription factor DREB, which can bind specific core sequence of DRE element A/GCCGAC, can induce the downstream functional genes which have response to salt, drought and low temperature. Based on the importance of DREB in signaling, this dissertation analyzed the response of DREB1 gene to abiotic stresses using CkDREB1 overexpression transgenic lines, and the results showed: 1. Three version of full-length DNA sequence of CkDREB were obtained from Caragana Korshinkii Kom. All the ORFs are 615bp in length. Sequence analysis showed that the conserved sequence such as PKK/RPAGRxKFxETRHP and DSAWR existed on the upstream and downstream of AP2 in CkDREB1 protein. Homology alignment indicated that CkDREB1 belonged to the A-1 subfamily of DREB protein family. 2. Northern blot assay showed that CkDREB1 could be induced