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干旱、高盐和病虫害严重影响了作物的产量和品质。ERF(ethylene responsive factor)转录因子是介导生物和非生物胁迫响应的重要调控因子。本实验室从小麦中克隆到抗逆相关ERF转录因子W17基因,本研究以植物表达载体pAHC25为基础,构建了含有反向重复序列的RNAi表达载体pAHC-W17RNAi。采用基因枪法45枪轰击小麦品种新春9号幼胚材料2 379个,共获得115株T0代再生植株,其中12株为阳性转基因植株。半定量RT-PCR分析表明,转基因植株中W17基因及其调控的下游基因的表达水平均显著下降;表型鉴定发现转基因小麦苗期发育受到抑制。本研究为深入分析小麦ERF基因的功能与作用机制,进而通过分子育种进行小麦抗性改良奠定了基础。
Droughts, high salt and pests and diseases have a serious impact on crop yield and quality. ERF (ethylene responsive factor) transcription factors are important regulators of biological and abiotic stress responses. Our lab cloned the anti-retrograde ERF transcription factor W17 gene from wheat. In this study, based on the plant expression vector pAHC25, we constructed the RNAi expression vector pAHC-W17RNAi with inverted repeat sequences. A total of 2 379 immature embryos of wheat cultivar Xinchun 9 were bombarded with 45 guns by using the gene gun method. A total of 115 T0 regenerated plants were obtained, of which 12 were positive transgenic plants. Semi-quantitative RT-PCR analysis showed that W17 gene and its downstream gene expression levels were significantly decreased in transgenic plants; phenotypic identification found that the development of transgenic wheat seedling stage was inhibited. This study laid the foundation for the further analysis of the function and mechanism of ERF gene in wheat and the improvement of wheat resistance by molecular breeding.