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基因转录调节是植物对非生物胁迫适应机制的一个重要方面,转录调节因子在胁迫信号转导途径中调节下游基因的表达,在建立植物对胁迫适应性过程中起到重要作用。DREB是功能多样的转录调节因子蛋白家族,家族成员在植物响应非生物胁迫方面扮演着重要角色。本研究以苜蓿MsDREB1基因为目的基因,分别把MsDREB1克隆到35S启动子与rd29A启动子之后,并把两种载体用农杆菌介导转入大豆基因组中,通过Southern检测转基因植株。15 d龄的幼苗在200 mmol·L~(-1)NaCl胁迫条件下,用RT-PCR分析基因不同时间的表达差异;并测定叶绿素、丙二醛、H_2O_2、SOD、相对根长及相对地上部分长度。结果表明:转MsDREB1基因在两种启动子驱动下均有一定耐盐能力,但存在差异。在非胁迫下35S启动子调控的MsDREB1为超量表达,而rd29A启动子调控MsDREB1表达量较低;在盐胁迫下,rd29A:MsDREB1表达量高于35S:MsDREB1的表达量;MsDREB1超量表达抑制植株正常生长。MsDREB1诱导表达耐盐性效果更明显,其植株脯氨酸含量、SOD活性均显著高于MsDREB1超量表达,而H_2O_2和MDA含量则显著低于MsDREB1超量表达。结果说明MsDREB1作为转录调节因子参与了植物的渗透调节,对植物的耐盐性具有贡献。该试验研究两种启动子调控的转MsDREB1基因大豆耐盐效果,为MsDREB1基因在大豆耐盐基因工程中的应用提供参考。
Transcriptional regulation is an important aspect of adaptation mechanism of plants to abiotic stresses. Transcription regulators regulate the expression of downstream genes in stress signal transduction pathways and plays an important role in establishing plant adaptability to stress. DREB is a versatile family of transcriptional regulator proteins that play important roles in the plant response to abiotic stresses. In this study, MsDREB1 gene of alfalfa was cloned into the 35S promoter and rd29A promoter respectively. The two vectors were transformed into soybean genome by Agrobacterium tumefaciens, and the transgenic plants were detected by Southern blot. The expression of chloroplast, malondialdehyde, H_2O_2, SOD, relative root length and relative root length were determined by RT-PCR under 15 mmol / L NaCl stress for 15 d. Part of the length. The results showed that the MsDREB1 gene had some salt tolerance ability under the two promoters, but there was a difference. The expression of MsDREB1 regulated by 35S promoter was overexpression and the expression of MsDREB1 regulated by rd29A promoter was lower under salt stress. The expression of MsDREB1 was higher than that of 35S: MsDREB1 under salt stress. MsDREB1 overexpression was inhibited Plants grow normally. The results showed that MsDREB1 induced more salt tolerance, and its proline content and SOD activity were significantly higher than that of MsDREB1 overexpression, while the content of H_2O_2 and MDA was significantly lower than that of MsDREB1 overexpression. The results indicate that MsDREB1 participates in osmoregulation of plants as a transcriptional regulator and contributes to the salt tolerance of plants. This study investigated the effect of two promoters on the salt tolerance of soybean transformed to MsDREB1 gene and provided a reference for the application of MsDREB1 gene in soybean salt tolerant genetic engineering.