地黄Aux/IAA家族基因RgIAA1的克隆和表达分析  

Molecular cloning and expression analysis of an Aux /IAA gene( RgIAA1) from Rehmannia glutinosa

作　　者：王丰青[1,2] 田云鹤[1] 李明杰[3] 杨金凤[1] 张宝[2] 林文雄[2] 陈新建[3] 张重义[3,2] 

机构地区：[1]河南农业大学农学院,河南郑州450002 [2]福建农林大学中药材GAP研究所,福建福州350002 [3]河南农业大学中药材研究所,河南郑州450002

出　　处：《中国中药杂志》2013年第23期4033-4039,共7页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(81274022;31271674);中国博士后科学基金项目(2013M541977);河南农业大学大学生创新实验计划项目(CX12007)

摘　　要：为了克隆地黄Aux/IAA家族基因RgIAA1,分析地黄RgIAA1的时空表达特性及对逆境胁迫的响应。以拟南芥AtIAA14的cDNA序列为探针在地黄转录组数据库中进行比对,应用生物信息学方法分析RgIAA1的序列特征及进化关系,以实时荧光定量PCR技术检测RgIAA1在地黄不同组织以及逆境胁迫下的表达量。结果表明,获得1条903 bp的cDNA序列,包含一个681 bp完整的开放阅读框(ORF),编码226个氨基酸,具有Aux/IAA家族蛋白的典型结构域和特征序列。RgIAA1在地黄不同组织中的表达呈差异表达,其中在地黄幼嫩叶片里表达强度最大,其次为茎,且随着叶片的伸展,RgIAA1表达强度不断降低。在连作时RgIAA1的表达量升高,NaCl和渍水胁迫下RgIAA1的表达量降低。实验首次克隆了地黄Aux/IAA家族基因RgIAA1,为阐明其在地黄生长发育和逆境胁迫中的分子功能奠定基础。To clone and analyze a member of the Auxin/indole-3-acetic acid（Aux/IAA） gene family, RglAA1, from Rehman- nia glutinosa. The transcriptional EST database of R. glutinosa was used to clone the new Aux/IAA gene by cDNA probe of AtIAA14. Bioinformatics was applied to analyze the sequence characteristics of RglAA1 protein and construct phylogenetic trees. Quantitative RT- PCR has been applied to detect the transcription level of RgIAA1 in seven tissues as well as in leaves under three stresses. The results showed that, the cDNA sequence of RglAA1 contains 903 bp was obtained. The open reading frame （ORF） of RgIAA1 was 681 bp en- coding 226 amino acids, which has typical structural domains and characteristic sequence of Aux/IAA family proteins. RglAA1 showed the highest expression level in unfolded leaf, followed by the stem. And the expression of RglAA1 was quickly decreased with leaf grow- ing up. The transcription level increased under continuous cropping conditions while it reduced both in salinity and waterlogging stres- ses. RglAAI, an Aux/IAA gene from R. glutinosa has been obtained for the first time, which can lay the foundation for further studies about its molecular function in development and responses to stress.

关 键 词：地黄 AUX IAA家族基因 序列特征 表达分析 

分 类 号：S567.239[农业科学—中草药栽培]