草业学报2019，Vol.28Issue(1)：128-137,10.DOI:10.11686/cyxb2018370
紫花苜蓿高温诱导启动子pMsMBF1c的克隆与功能分析
Cloning and function analysis of the high temperature inducible promoter pMsMBF1c in alfalfa (Medicago sativa)
摘要
Abstract
Plant thermotolerance is controlled by a complex and finely regulated network of transcription processes.High temperature inducible promoters can economically and efficiently activate or shut down the expression of key genes involved in regulating thermotolerance.Knowledge of these promoters is therefore important in plant functional genomics research and the molecular breeding industry.In the present study, a 1748bp sequence upstream of MsMBF1c, which is a c andidate gene involved in thermotolerance in alfalfa, was isolated using a digestion ligation PCR method, based on its coding sequence.Bioinformatics analysis showed that two conserved motifs representing the HSE and GATA binding sites involved in thermotolerance regulation existed in the MsMBF1c promoter region.There were 5 abscisic acid response cis-elements (ABRE, MYB2, MIC2, CBF and DPBF) and 2 MYB protein binding motifs in the MsMBF1c promoter region, which suggested MsMBF1c might regulate other abiotic stress response in addition to thermotolerance.A vector of pBI121-MsMBF1c::GUS was constructed and transformed into wild type Arabidopsis.The expression of GUS and AtMBF1c genes, was examined by qRT-PCR, and found to be up-regulated 5.4 and 4.8 fold, respectively, after heat stress.At the same time, histochemical staining analysis also showed the MsMBF1c promoter to be markedly induced by high temperature.To summarize, the MsMBF1c promoter was isolated and transformed into Arabidopsis, and was proved to be heat inducible by bioinformatics and gene expression data, and by histochemical staining analysis.This information provides technical and theoretical support for further investigation of heat regulation mechanisms and breeding of heat resistant alfalfa varieties.关键词
高温/MBF1c/紫花苜蓿/转基因/启动子Key words
high temperature/MBF1c/alfalfa/transgene/promoter引用本文复制引用
李小冬,莫本田,牟琼,娄芬,陈文贵,陈光吉,张瑜,韩永芬..紫花苜蓿高温诱导启动子pMsMBF1c的克隆与功能分析[J].草业学报,2019,28(1):128-137,10.基金项目
贵州省农业科学院专项基金(黔农科院院专项[2013]03),贵州省科学技术基金(黔科合J字[2015]2080号),省科研机构服务企业行动计划项目(黔科合平台人才[2016]5713)和贵州省农业科学院青年基金(黔农科院青年基金[2017]18)资助. （黔农科院院专项[2013]03）
