中间锦鸡儿CiDREB1C基因增强转基因拟南芥抵抗非生物胁迫的能力  

CiDREB1C Gene from Caragana intermedia Enhances Abiotic Stress Tolerance of Transgenic Arabidopsis thaliana

作　　者：刘坤[1] 王光霞[1] 王瑞刚[1,2] 李国婧 LIU Kun;WANG Guang-Xia;WANG Rui-Gang;LI Guo-Jing(College of Life Sciences,Inner Mongolia Agricultural University,Hohhot 010018,China;Inner Mongolia Key Laboratory of Plant Stress Physiology and Molecular Biology,Hohhot 010018,China)

机构地区：[1]内蒙古农业大学生命科学学院,呼和浩特010018 [2]内蒙古自治区植物逆境生理与分子生物学重点实验室,呼和浩特010018

出　　处：《农业生物技术学报》2018年第10期1688-1697,共10页Journal of Agricultural Biotechnology

基　　金：内蒙古自治区科技创新引导资金(No.KCBJ2018012);呼和浩特市科技计划重大专项(2017-社-重-4);国家自然科学基金(No.31060105)

摘　　要：中间锦鸡儿(Caragana intermedia)是广泛分布于我国西北干旱、半干旱荒漠区的防风固沙先锋树种,具有极强的抗逆性和饲用价值。本课题组从前期建立的干旱处理的中间锦鸡儿转录组数据库中分离到1个脱水响应元件结合蛋白(dehydration responsive element binding protein, DREB)转录因子编码基因片段,为了检测CiDREB1C基因的抗旱和抗冷能力,本研究首先利用qRT-PCR技术检测中间锦鸡儿中CiDREB1C基因在冷和干旱胁迫下的表达水平,发现均有上升(P<0.01)。为了验证该基因的功能,本研究利用PCR技术获得了CiDREB1C基因全长(GenBank No. MG748598),并将其构建入植物过表达载体pCanG-HA,通过农杆菌介导法转化野生型拟南芥(Arabidopsis thaliana),获得了超表达CiDREB1C的转基因拟南芥纯合体。不同胁迫处理结果显示:甘露醇处理下,转基因拟南芥丙二醛的含量显著低于野生型(P<0.01),叶绿素含量明显高于野生型(P<0.01),表现出明显的抗旱表型;冷胁迫处理后,过表达株系叶绿素含量明显高于野生型(P<0.01);黑暗处理下,过表达株系叶片衰老延迟,细胞死亡较野生型少,叶绿素含量显著高于野生型(P<0.01)。上述结果说明,CiDREB1C基因提高了转基因拟南芥对多种胁迫的耐受性。本研究结果为进一步研究CiDREB1C基因在抗逆中的功能及中间锦鸡儿的抗逆分子作用机理提供依据。Caragana intermedia is a pioneer species for wind-controlling and sand-fixing and distributes widely in arid and semi-arid desert area of north-west China. It has extremely strong resistance to stress and is a good feeding stock. A dehydration responsive element binding protein（DREB） transcription factor encoding gene has been isolated from the dought-treated transcriptome data set. In order to detect the drought and cold resistance of CiDREB1C, expression level of CiDREB1C gene under drought and cold in C. intermedia was detected by qRT-PCR. In order to identify the gene＇s function, the full length CiDREB1C（GenBank No.MG748598） were obtained through PCR, and was cloned into the expression vector pCanG-HA. Transgenic Arabidopsis thaliana had been obtained by Agrobacterium mediated genetic transformation, and the homozygous transgenic A. thaliana had been screened. The results showed that under mannitol treatment, the overexpression lines accumulated less malondialdehyde（P〈0.01）, and had more chlorophyll compared with the wild type, showed obvious drought resistance phenotype. After cold treatment, the overexpression lines showed a significant increase in chlorophyll content compared with the wild type as well（P〈0.01）. Under darkness treatment, the leaves of overexpression lines were greener than those of wild type, the cell death was less than that of wild type and the chlorophyll content was significantly higher than that of wild type（P〈0.01）. These results suggested that overexpression of CiDREB1C could significantly increase tolerance of the transgenic A. thaliana to drought, cold and dark stress.The results of this research provide a theoretical basis for further analysis the function of CiDREB1C gene in stress resistance and the molecular mechanism of stress resistance of C. intermedia.

关 键 词：脱水响应元件结合蛋白(DREB) 转基因拟南芥 非生物胁迫 中间锦鸡儿 

分 类 号：S793.3[农业科学—林木遗传育种] Q786[农业科学—林学]