作 者 ：黄 莹,徐志胜,王 枫,李梦瑶,马 静,熊爱生*

期 刊 ：西北植物学报 2015年 1期 页码：30~36

关键词：胡萝卜；DcICE1转录因子；基因克隆；非生物胁迫；表达分析；

Keywords:Daucus carota, DcICE1 transcription factor, gene cloning, abiotic stress, expression analysis,

摘 要 ：植物ICE1基因是调控CBF基因表达的上游调控因子,在植物抵抗逆境胁迫中具有重要的作用。该实验以2个胡萝卜品种 ‘黑田五寸’和‘君川红’为实验材料,分别克隆出DcICE1转录因子基因,并通过荧光定量PCR方法测定了4种不同逆境胁迫下(4 ℃低温、38 ℃高温、0.2 mol·L^-1 NaCl及200 g·L^-1 PEG)DcICE1基因的表达情况,探讨DcICE1转录因子在植物抵抗非生物胁迫下的功能。序列分析显示,该基因全长1 458 bp,编码485个氨基酸。2个胡萝卜品种的DcICE1基因在核苷酸水平上有2个位点的差异,分别为第139位的G/A和第475位的A/G,导致编码的氨基酸在第47位的E/K和第159位的N/D差异,该差异可能与DcICE1转录因子在2个不同品种应对逆境胁迫下的响应不同有关。胡萝卜DcICE1转录因子有一个保守的ICE1功能结构域。实时定量PCR检测DcICE1基因在不同逆境胁迫下的响应表明,低温(4 ℃)处理能明显地诱导DcICE1基因的表达,但盐(0.2 mol·L^-1 NaCl)和干旱(200 g·L^-1 PEG)处理下的诱导效果均不明显。

Abstract:In higher plant,ICE1 involved in the response to abiotic and biotic stresses by regulating the expression of CBF genes.The cDNA sequence of DcICE1 transcription factor was cloned from carrot cultivars ‘Heitianwucun’ and ‘Junchuanhong’ using RT-PCR method,respectively.In order to analyze the response of DcICE1 gene to abiotic stress,the expression profiles of DcICE1 gene in four different stresses(4 ℃,38 ℃,0.2 mol·L^-1 NaCl,200 g·L^-1 PEG) were detected by quantitative real-time PCR.Sequence analysis indicated that the DcICE1 gene contained 1 458 bp,encoding 485 amino acids.There were two nucleotide sites and two amino acid residue differences between two cultivars(G/A in site 139 and A/G in site 475;E/K in site 47,N/D in 159).The different amino acid residues may be involved into the different response in two carrot cultivars encountered the abiotic stresses.The DcICE1 transcription factor has a conserved domain.Quantitative real-time PCR detected the expression profiles of DcICE1 gene under abiotic stresses.The results showed the expression of DcICE1 gene significantly increased when exposed to 4 ℃,while the expression profiles of DcICE1 gene was not obvious after treated by salt and PEG in the two carrot cultivars.