转CP基因大豆植株抗大豆花叶病毒的鉴定与筛选
时间:2018-12-10 21:30 来源:毕业论文 作者:毕业论文 点击:次
| 摘要：本研究是在前人的研究基础上展开，受体材料是华春6号大豆品种，其转基因材料是通过农杆菌介导子叶节转化方法将大豆花叶病毒的CP基因的保守序列导入大豆基因组中（携带抗草丁膦除草剂基因），借助RNA干扰（RNAi）机制引发RNA沉默（RNA silencing），即序列特异性的转录后基因沉默（Post-transcriptional gene silencing，PTGS），从而诱发RNA介导的抗性（RNA-mediated resistance），即源于病原诱导的抗性（Pathogen-derived resistance，PDR），最终培育出对SMV具有广谱、持久抗性的转基因大豆新材料。 本研究的开展是从4个转基因T2代家系开始，利用除草剂涂抹、PCR检测、PAT/bar转基因检测试纸三种方法对基于外源基因CP的T2转基因材料进行鉴定，结果显示，4个T2代家系衍生的13个T3代家系的个体材料均表现为转基因阳性；通过接种SC3、SC7、SC15和SC18四个SMV株系并通过田间表现进行抗性筛选得到，转基因材料接种各种病毒的家系平均发病等级均低于非转基因对照的发病等级。从中选择8个家系（覆盖四种病毒类型），在接种SMV3周后和5周后取样进行DAS-ELISA实验，结果表明，在转基因植株体内检测不到SMV病毒存在，而非转基因对照在两个时期病毒含量很高。31410 本研究旨在对转基因抗病育种技术提供一定的借鉴，其后续工作着眼于侧重筛选可稳定遗传的高抗SMV的转基因大豆纯合家系；并对转基因大豆进行农艺性状调查和营养成分检测，为转基因大豆品种的推广应用奠定基础。 毕业论文关键词：大豆；大豆花叶病毒；RNA干扰；鉴定和筛选 Identification and Screening of Soybean Mosaic Virus with CP Gene Soybean Plant Abstract：This study is based on the study of predecessors, the receptor material is Huachun 6 soybean varieties,and its transgenic material was introduced into the soybean genome by the Agrobacterium tumefaciens-mediated cotyledon transformation method. Based on the RNAi mechanism, the RNA silencing, otherwise known as post-transcriptional gene silencing (PTGS) can be induced in the transgenic plants. Transgenic soybeans with broad-spectrum and persistent SMV-resistance can be achieved by conferring RNA-mediated resistance, namely Pathogen-derived resistance (PDR). In this study, T2 transgenic materials based on exogenous gene CP were identified by four methods of herbicide application, PCR detection and PAT / bar transgenic test paper. The results showed that the inpidual materials of 13 T3 generation families derived from four T2 generation families showed transgenic positive.Through the field performance of the material which were inoculated with SC3, SC7, SC15 and SC18 four SMV lines,We can know that The average incidence level of genetically modified materials was lower than that of non-transgenic controls.Eight families (covering four virus types) were inoculated with SMV, and DAS-ELISA was performed after 3 weeks and 5 weeks.The results showed that SMV virus was not detected in transgenic plants, but non-transgenic materials were highly viral in both periods. The aim of the study was to provide some reference for transgenic resistance breeding technology,The follow - up work focused on screening for homozygous families with high resistance to SMV, which can be stably inherited;And the agronomic traits investigation and nutrient analysis of transgenic soybean, lay the foundation for the popularization and application of transgenic soybean varietie. Key words: Soybean; soybean mosaic virus; RNA interference; identification and screening 目 录 摘要1 关键词1 Abstract2 Key words2 引言（或绪论）3 1材料与方法3 1.1 材料 3 1.2 方法 4 1.2.1 汁液摩擦接种法繁殖、活化SMV4 1.2.2 转基因植株的鉴定4 1.2.3 转基因植株抗性筛选5 (责任编辑：qin) |