叶脉分化发育关键基因COV1在柠条锦鸡儿中的克隆和表达分析  

Cloning and Expression Analysis of COV1,a Key Gene in the Differentiation and Development,From Caragana korshinskii Kom.

作　　者：高丽芳 孙莹莹[1] 周文菲 白娟[1] 龚春梅[1] 

机构地区：[1]西北农林科技大学生命科学学院,陕西杨凌712100

出　　处：《核农学报》2018年第1期40-47,共8页Journal of Nuclear Agricultural Sciences

基　　金：国家自然科学基金面上项目(31070538;31370599)

摘　　要：COV1是植物叶脉发育中的关键基因。为探究随着干旱加剧COV1在调控柠条锦鸡儿维管系统尤其是叶脉分化发育中是否起关键作用,从非模式植物柠条锦鸡儿叶样转录组测序数据库中筛选到一条COV1序列,通过与NCBI数据库中已知序列Blast比对,设计克隆和定量引物,并通过生物信息学方法分析克隆得到的基因序列。克隆基因长为946 bp的序列,其中开放阅读框为777 bp,编码258个氨基酸,推测该蛋白分子量为28.84 KD,等电点为5.46,是一种疏水蛋白。MEGA 5生物信息学软件构建聚类树发现,柠条锦鸡儿中COV1与豆科植物中的模式植物蒺藜苜蓿COV1亲缘关系最近。利用RTq PCR方法对柠条叶样干旱胁迫下该基因表达水平进行定量分析,结果表明,随着干旱胁迫的加剧,该基因在柠条锦鸡儿叶样中锦鸡儿的表达量呈下降趋势,说明柠条锦鸡儿在干旱胁迫加剧下,COV1基因的表达抑制可能与维管系统发育相关。本研究结果为进一步明确干旱胁迫下促进柠条锦鸡儿维管系统发育的分子机制提供了理论依据。COV1 is a key gene in leaf vein development. To explore the vital function of COV1 in regulating the development of Caragana korshinskii Kom. vascular system,especially in the development of vein differentiation under the increasing drought, a COV1 encoding gene fragment was isolated from transcriptome sequencing database of Caragana korshinskii Kom.,and the parthal length c DNA was cloned by RT-PCR. Sequencing results showed that the parthal c DNA length was 946 bp,with an open reading frame of 777 bp. It encoded a protein composed of 258 amino acids. The putative protein molecular weight was 28. 84 KD,and isoelectric point was 5. 46. The deduced protein is strongly hydrophobic. Bioinformatics analysis showed that the COV1 gene in Caragana korshinskii Kom. was most closed to COV1 in the leguminous model plant Medicago truncatula Kom.. RT-q PCR analysis showed that the transcript of COV1 was inhibited under drought stress. These results indicated that the suppression of COV1 gene expression under drought stvess is associated possibly with vascular system development. This further clarifying molecular mechanism of developed vascular system in Caragana korshinskii Kom. under the drought stress.

关 键 词：COV1 柠条锦鸡儿 干旱胁迫 维管系统 

分 类 号：Q943.2[生物学—植物学] S793.3[农业科学—林木遗传育种]