中文摘要：

该研究在生物信息学分析的基础上,克隆玉米胚胎发生后期丰富蛋白基因（MGL3）的启动子序列（pMGL3）,进行非生物逆境应答元件分析以及实时定量PCR验证其非生物逆境胁迫响应特性,构建了pMGL3启动子驱动报告基因（GUS）表达载体,基因枪法转化玉米愈伤组织,通过GUS染色验证pMGL3启动子在非生物逆境胁迫下的驱动活性。再根据启动子序列分析结果,去除不同的顺式作用元件,构建不同长度pMGL3启动子驱动报告基因GUS表达载体,农杆菌介导法转化烟草叶盘,以确定pMGL3启动子的最短活性序列。结果显示：pMGL3启动子长1 554bp,存在多种与非生物逆境胁迫应答相关的调控元件,在干旱、高盐、低温胁迫及脱落酸、乙烯诱导下驱动MGL3基因增量表达,用以驱动GUS基因转化玉米愈伤组织,在高渗、高盐、低温胁迫及脱落酸诱导下具有驱动活性,且截短至325bp仍可保持驱动活性。研究表明,pMGL3启动子的确有非生物逆境诱导启动活性,进一步验证其作用机理后可运用于玉米抗逆转基因研究。

英文摘要：

It has important significance for transgenic resistance research to clone and validate endogenous inducible promoters in response to abiotic stresses, and use to construct inducible expression vectors. In this study,the sequence of the late embryogenesis abundant late gene （MGL3） promoter （pMGL3） was cloned from maize according to bioinformatics analysis. After analysis for abiotic stress-responsive elements and abiotic response by quantitative real-time PCR,we used this sequence to construct expression vector of the reporter gene GUS, and used to transform maize call/ by biolistic method. The promotion activity of the pMGL3 promoter under abiotic stress was validated by GUS staining. In addition, the different cis-acting elements was removed according to promoter sequence analysis, used to construct expression vectors of the reporter gene GUS, and transform tobacco discs by Agrobacterium mediation, in order to determine the shortest active sequence of the pMGL3 promoter. The results showed that the pMGL3 promoter is 1 554 bp long,containing multiple regulatory elements in response to abiotic stress. The expression of the MGL3 gene is increased under the stress of drought, high salt and low temperature, and induction of abscisic acid and ethylene. The calli transformed by the GUS gene under control of the pMGL3 promoter showed promo- tion activity under high osmotic, high salt,low temperature stresses,and abscisic acid induction. However,it kept promotion activity when truncated to as short as 325 bp. These results indicated that the pMGL3 promoter has promotion activity in response to abiotic stresses,and can be applied to maize transgenic studies for abiotic tolerance after further validation for its mechanism.