木薯低温诱导基因MeLTI6A的启动子的克隆与序列分析  

Cloning and Sequence Analysis of Low Temperature Inducible Gene MeLTI6A Promoter from Manihot esculenta

作　　者：黎娟华[1] 孙海彦[1] 阮孟斌[1] 赵平娟[1] 曾长英[1] 彭明[1] 

机构地区：[1]中国热带农业科学院热带生物技术研究所农业部热带作物生物学与遗传资源利用重点实验室,海南海口571101

出　　处：《热带作物学报》2013年第11期2164-2171,共8页Chinese Journal of Tropical Crops

基　　金：国家973计划项目(No.2010CB126602);中央级公益性科研院所基本科研业务费专项项目(No.ITTBB110103)

摘　　要：MeLTI6A(Manihot esculenta low temperature inducible 6A)是木薯低温干旱诱导基因,本研究从MeLTI6A的序列出发,利用电子克隆设计引物进行PCR扩增的方法获得该基因的启动子,其序列共1 304 bp。生物信息学分析发现该启动子中具有真核生物典型的核心启动子区(TATA-box和CAAT-box),并利用α-互补,蓝白斑筛选原理验证了该启动子核心序列具有活性;该启动子具有与干旱胁迫相关的激素类(如脱落酸、乙烯)的响应元件和逆境胁迫(如低温、干旱胁迫)的响应元件;还具有与木薯组织特异表达相关的调控元件和其它光响应元件;并通过Real time PCR检测了低温胁迫(4℃)下的木薯组培苗中MeLTI6A的表达变化,说明了该启动子区的低温胁迫顺式作用响应元件可能调节MeLIT6A在低温胁迫下的表达。这些说明木薯的MeLIT6A基因可能是通过对干旱胁迫激素信号响应以及逆境胁迫响应起作用,使木薯获得一定的抗胁迫的能力,同时还可能参与了木薯相关组织发育过程的调控。本研究有利于对MeLTI6A基因抗逆境胁迫功能的理解,为探索木薯高效抗逆的分子机制作初步研究。MeLTI6A （Manihot esculenta low temperature inducible 6A） is a low temperature inducible and drought resistance gene of Cassava.In this study,the promoter of MeLTI6A,1 304 bp,was cloned by designing primers for PCR amplification from electronic cloning based on the sequence of MeLTI6A.Bioinformatics analysis found that the promoter had the typical eukaryotic promoter core region （TATA-box and CAAT box）,and the activation of the promoter core region was validated by using the alpha-complementary,blue and white spot screening principle.The promoter had drought stress related hormones responsive elements （abscisic acid,ethylene e.g.） and adversity stress related responsive elements （low temperature,drought stress e.g.）.The promoter also had cassava tissue specific expression regulation elements and other light responsive elements.MeLTI6A expression changes of cassava somaclone under low temperature stress （4 ℃） were detected by using real time PCR.It proved that the MeLIT6A expression patterns under low temperature stress were might regulated by the low temperature stress cisacting responsive elements of the promoter.All that suggested that the function of MeLIT6 might contribute to the cassava adversity stress resistance via the drought stress hormone signal response and adversity stress response.This study was good for us to understand the adversity stress resistant function of MeLTI6A,and it was a preliminary exploration for the molecular mechanism study of the high-efficient adversity stress resistance of cassava.

关 键 词：木薯 启动子 木薯低温诱导基因MeLTI6A 启动子序列分析 

分 类 号：S533[农业科学—作物学]