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[目的]研究从烟草中克隆的1个受水杨酸和茉莉酸甲酯诱导表达的新的糖基转移酶基因(sm-Ngt)启动子部分缺失片段在烟草中的表达。[方法]以转sm-Ngt5个不同长度的5’端缺失的启动子与Gus基因融合植物表达载体的T1代转基因植株为材料,用茉莉酸甲酯(MeJA)和水杨酸(SA)处理16h后分别进行GUS组织化学染色和荧光定量法测定GUS酶活性,分析水杨酸和茉莉酸甲酯对sm-Ngt5个不同长度的5’端缺失的启动子表达的影响。[结果]在5个不同缺失片段启动子的转基因T1代生长30d的植株中,转-220～0bp片段的GUS染色最少,转-524～0bp及-468～0bp片段的染色最深。在没有MeJA和SA诱导处理时,-524～0bp和-468～0bp2个启动子片段启动的GUS活性最高,远高于-1150～0、-800～0、-220～0bp片段的活性,并且不是由于基因拷贝数而引起的(Southern杂交结果);-800～0bp启动子片段启动的GUS活性受到MeJA和SA双重诱导,-1150～0bp启动子片段启动的GUS活性受到MeJA的诱导。[结论]在sm-Ngt启动子的-524～-220bp存在提高启动子活性调控元件,-1150～-524bp存在抑制启动子活性的序列,并且存在MeJA和SA双重诱导启动子活性调控元件。
[Objective] The research aimed to study the expression of a partial deletion fragment of sm-Ngt promoter in tobacco induced by one of salicylic acid and methyl jasmonate cloned from tobacco. [Method] T1 generation transgenic plants with five different lengths of sm-Ngt 5-terminal deletion promoter and Gus gene fusion plant expression vector were used as material and treated with methyl jasmonate (MeJA) and salicylic acid (SA) GUS histochemical staining and fluorescence quantitative detection of GUS activity were performed 16 h later. The effects of salicylic acid and methyl jasmonate on the expression of sm-Ngt 5-terminal deletion promoters were analyzed. [Result] The transgenic plants with 5 different deletion promoters grew 30d in transgenic T1 generation. The transfection of -220 ~ 0bp showed the least staining of GUS, and the transfection of -524 ~ 0bp and -468 ~ 0bp showed the deepest staining. In the absence of induction treatment with MeJA and SA, the promoter activity of two promoters, -524 ~ 0bp and -468 ~ 0bp, was the highest, much higher than those of -1150 ~ 0, -800 ~ 0 and -220 ~ 0bp, Was not due to gene copy number (Southern hybridization results); - GUS activity initiated by 800 ~ 0bp promoter fragment was induced by both MeJA and SA, and GUS activity initiated by -1150 ~ 0bp promoter fragment was induced by MeJA. [Conclusion] There was a promoter inhibitory activity promoter in -524 ~ -220bp of sm-Ngt promoter and -1150 ~ -524bp promoter. The regulatory elements of MeJA and SA dual promoter activities existed.