蒺藜苜蓿转录因子MtMYB的克隆、表达分析及拟南芥转化  

Cloning and Expression Analysis of Transcription Factor MtMYB and Transformation of Arabidopsis thaliana

作　　者：李殷睿智 冷暖 穆致远 滕珂[1] 王焕英 晁跃辉[1] Li Yinruizhi;Leng Nuan;Mu Zhiyuan;Teng Ke;Wang Huanying;Chao Yuehui(Turfgrass Research Institute of Beijing Forestry University,Beijing,100083)

机构地区：[1]北京林业大学,草坪研究所,北京100083

出　　处：《分子植物育种》2020年第9期2769-2775,共7页Molecular Plant Breeding

基　　金：国家自然科学基金青年科学基金项目(31601989);深圳市科技计划项目(JCYJ20160331151245672,JSGG-20160229155434792);中国博士后基金(2017M620677);北京市博士后基金(2017-ZZ-087)共同资助。

摘　　要：MYB转录因子家族基因具有保守的DNA结合域,是植物最大的转录因子家族之一,广泛参与植物的代谢调控,在植物生长发育和胁迫响应等方面发挥重要作用。为改良品种性状和提高生产实践能力,本研究采用RT-PCR法克隆蒺藜苜蓿MYB转录因子基因MtMYB,其编码区长255 bp,编码84个氨基酸。生物信息学分析表明,该基因编码蛋白与其他物种MYB蛋白具有很高的同源性,其中与红三叶同源性最高。荧光定量结果表明,MtMYB基因在蒺藜苜蓿叶片中的表达量最高。MtMYB基因在外源IAA、6-BA的诱导下,总体都呈现先上升后下降的趋势。同时本研究成功构建35S::MtMYB的植物表达载体,并通过转基因技术将其转化至拟南芥中,PCR检测表明,MtMYB已成功整合至转基因拟南芥基因组中。RT-PCR检测表明,MtMYB能够在转基因拟南芥中正常转录。本研究为进一步探明MtMYB基因的功能提供了数据参考和实验材料。MYB transcription factors with a conservative DNA binding domain in common is one of the largest transcription factor families in plants.It is widely involved in plant metabolic regulation,and plays an important role in plant development,stress response and so on.In order to improve variety traits and productivity practices,the transcription factor MtMYB from Medicago truncatula was cloned by RT-PCR.The coding domain was 255 bp,encoding 84 amino acids.Bioinformatics analysis showed that protein coding by this gene had high homology with other species MYB protein,especially Trifolium prateose L..Quantitative real-time PCR showed that expression of MtMYB gene was highest in leaves.Under the induction of exogenous IAA and 6-BA,expression of MtMYB increased firstly and then decreased as a whole.At the same time,the plant over-expression vector of 35S::MtMYB was successfully constructed,transferring into Arabidopsis by transgenic technology.PCR identification of total DNA from transgenic Arabidopsis showed that MtMYB had been integrated into transgenic Arabidopsis genome.RT-PCR identification of total RNA showed that MtMYB could transcribe normally in transgenic Arabidopsis thaliana.This study provides data reference and experimental materials for further study of the function of MtMYB gene.

关 键 词：蒺藜苜蓿(Medicago truncatula) MtMYB 表达分析 拟南芥(Arabidopsis thaliana) 

分 类 号：Q943.2[生物学—植物学]