【作者】 刘永光；

【导师】 车代弟；

【作者基本信息】 东北农业大学 ， 园林植物与观赏园艺， 2006， 硕士

【摘要】 园林植物多进行保护地栽培,高温高湿的局部小气候使园林植物更容易感染多种病害,而一旦染病则会严重影响园林植物的观赏价值,造成经济上的重大损失。一方面,园林植物抗病育种周期长,能够用做抗病育种的园林植物种质资源贫乏,短时间内园林植物抗病育种难以有所大的突破;另一方面,日益蓬勃发展的园林业迫切需要能够抵抗多种病害的广谱抗病商品植株。系统获得性抗性(Systematic acquired resistance, SAR)是植物抵御病原菌侵染的最有效手段,我们可以利用SAR信号传导途径中的关键功能基因来启动园林植物自然防御系统,达到广谱抗病的目的。NPR1基因在植物的SAR发生中起着重要的作用,通过NPR1基因的过量表达可以诱导植物SAR的起始,使植物产生对多种病原菌的广谱抗病性。TGA2转录因子能直接和多个病程相关基因启动子结合感应SA的调节元件结合,能够增强病程相关蛋白的表达水平,特异的诱导植物产生SAR,从而启动植物的广谱抗病。本研究主要研究成果如下:1.通过反转录PCR,从拟南芥中克隆出广谱抗病基因NPR1。测序结果显示所克隆NPR1基因全长1782bp,与NCBI所登陆的基因100%同源。2.通过反转录PCR,从拟南芥中克隆出能够诱导广谱抗病的TGA2转录因子。测序结果显示所克隆TGA2基因全长993bp,与NCBI所登陆的基因99.8%同源,发生了两个碱基的置换,但所翻译氨基酸没有变化。3.将NPR1基因正向插入中间载体的CaMV35S启动子和T-nos终止子之间,构建出p35ST-NPR1植物表达载体。4.将TGA2转录因子正向插入中间载体的CaMV35S启动子和T-nos终止子之间,构建出p35ST-TGA2植物表达载体。5.将NPR1基因正向插入到pMXB10蛋白表达载体中,构建出pMXB10-NPR1蛋白表达载体。6.将TGA2转录因子正向插入到pMXB10蛋白表达载体中,构建出pMXB10-TGA2蛋白表达载体。更多还原

【Abstract】 The horticulture plants usually are cultivated in the greenroom where are easy to cause many bacterial and fungal diseases. Once were infected by the all kinds of diseases, the horticulture plants’value of view and admire was seriously affected and the economy value was seriously reduced.Systematic acquired resistance (SAR) is the most effective way in plant resistance to pathogens. Using the key gene in systematic acquired resistance (SAR) signal transduction, we can introduced the plants own disease-resistance which can gain a broad-spectrum disease resistance. NPR1 genes play an important role in the plant SAR. Over-expressing NPR1 gene show high resistance to many pathogens and the transgenic plants with over-express NPR1 gene also enhanced its resistance to a broad-spectrum disease. They found: In the downstream of signals transmission channels, there are a lot of same transcription factors which join in the defensive reaction induced by differently resistant gene. The TGA2 transcription factor can directly combine with most of pathogens-related gene and enhance the express level of pathogens-related protein. TGA2 transcription factor synergized with NPR1 gene can induce the plants SAR which can specially induce the pathogenesis-related protein and achieved the efforts of a broad-spectrum disease resistance.The main results were summarized as follows:1. NPR1 gene was cloned from Arabidopsis thaliana by RT-PCR. The result of sequence analysis showed that the length of gene was 1782bp .Compared with the published sequence in the NCBI they are 100% homologous.2. TGA2 transcription factor was cloned from Arabidopsis thaliana by RT-PCR. The result of sequence analysis showed that the length of transcription factor was 993bp .Compared with the published sequence in the NCBI they are 99.8% homologous. There were two nucleic acids permutations, but permutations were out of the function area of transcription factor.3. In the experiment, we insert the NPR1gene into the position which is between CaMV35S promoter and T-nos terminator of media expression vectors. We achieved the p35ST-TGA2 plant expression vectors4. In the experiment, we insert the TGA2 transcription factor into the position which is between CaMV35S promoter and T-nos terminator of media expression vectors. We achieved the更多还原