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| 乙烯应答因子W17基因RNAi表达载体的构建及小麦遗传转化 |
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闵东红1, 陈阳1,2, 霍冬英1, 胡笛1, 赵月1,2, 徐兆师2, 张小红1, 李连城2, 陈明2, 马有志2
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| (1.西北农林科技大学 农学院/旱区作物逆境生物学国家重点实验室, 陕西杨凌 712100;2.中国农业科学院 作物科学研究所/国家农作物基金资源和基因改良重大科学工程/农业部作物遗传改良与育种重点开放实验室, 北京 100081) |
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| 摘要: |
| 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。 |
| 关键词: 小麦 转基因 遗传转化 ERF(乙烯应答因子) RNAi |
| DOI：10.11841/j.issn.1007-4333.2013.01.003 |
| 投稿时间：2012-09-07 |
| 基金项目:国家自然科学基金项目(31171546); 转基因生物新品种培育重大专项(2011ZX08002-002) |
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| Construction of the RNAi vector of ethylene responsive factor W17 gene and genetic transformation in wheat |
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MIN Dong-hong1, CHEN Yang1,2, HUO Dong-ying1, HU Di1, ZHAO Yue1,2, XU Zhao-shi2, ZHANG Xiao-hong1, LI Lian-cheng2, CHEN Ming2, MA You-zhi2
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| (1.College of Agronomy, Northwest A&F University/State Key Laboratory of Arid Regions Crop Adversity Biology, Yangling 712100, China;2.Institute of Crop Science/The National Key Facility for Crop Gene Resources and Genetic Improvement/Key Laboratory ofCrop Genetics and Breeding of Ministry of Agriculture, Chinese Academy of Agricultural Sciences, Beijing 100081, China) |
| Abstract: |
| Drought,high salinity,insect pest,and disease have adverse effect on the yield and quality of crops.Ethylene responsive factors (ERFs) are involved in plant response to abiotic and biotic stress.In this study,according to the sequence of an ERF family gene W17 from wheat,the W17 RNAi expression vector Pahc25-W17RNAi containing inverted-repeat sequences of W17 was constructed and transformed into wheat variety Xinchun 9 using particle bombardment.In total,2379 embryos were bombarded.A total of 115 regenerated plants with resistance to Bialaphos were obtained.Among them,12 transgenic lines with Bar gene were identified.Semi-quantitative RT-PCR showed that the expression of W17 gene and its downstream genes decreased significantly.Meanwhile,phenotype appraisal found that the growth of transgenic wheat in seedling stage was also inhibited.This study is useful for deep analysis of the function of ERF genes and preliminary investigation into the mechanism of ERF proteins;furthermore,it establishes the foundation for improving the wheat resistance by molecular breeding. |
| Key words: wheat transgenic genetic transformation ERF(ethylene responsive factors) RNA interference |
