CfTOM1抗病毒转基因辣椒的遗传分析
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摘要
拟南芥AtTOM1基因是拟南芥中系统获得的支持烟草花叶病毒复制的寄主因子基因。为了鉴定辣椒的一个与拟南芥AtTOM1基因同源的CfTOM1基因的功能，本实验室构建了一个以CfTOM1为靶基因的RNA干涉辣椒表达载体pBICRNAi3，并用根癌农杆菌介导法转化辣椒“天等一号”，共获得33株抗性再生苗。对T_0、T_1和T_2代植株进行PCR检测、PCR产物测序和PCR-Southern杂交分析，证实了这些再生苗为转基因植株。对T_2代植株分别进行烟草花叶病毒、黄瓜花叶病毒和番茄花叶病毒接种实验，结果表明转基因植株对三种病毒的抗性显著增强，证明辣椒CfTOM1基因是支持病毒复制与维持相关的寄主因子，其作用机制很可能与拟南芥的AtTOM1基因相似。
The Arabidopsis thaliana AtTOM1 is a host factor gene involved in the replication of tobacco mosaic virus(TMV).To identify the function of CfTOM1, an Arabidopsis thaliana AtTOM1 homolog gene in Capsicam frutoscens, an RNAi constrant vector pBICRNAi3 for silencing CfTOM1 was constructed and transformed to pepper cultivar Tiandeng No. 1 by Agrobacterium tumefaciens-infection. Thirty three selection marker-positive regenerated lines were detained.To lines and offsprings of T_1 and T_2 lines were further confirmed by PCR Sequencing and Southern analysis of the transgene to select the transgenic lines.T_2 lines were assayed for resistance to TMV , cucumber mosaic virus (CMV) and tomato mosaic virus (ToMV). It turned out that the transgenic pepper expressed significantly enhanced disease resistance to these three viruses. The results suggest that CfTOM1 is a host factor gene possibly involved in the replication of virus in pepper, likely throught the similar mechanism as AtTOM1 in Arabidopsis.
The Arabidopsis thaliana AtTOM1 is a host factor gene involved in the replication of tobacco mosaic virus(TMV).To identify the function of CfTOM1, an Arabidopsis thaliana AtTOM1 homolog gene in Capsicam frutoscens, an RNAi constrant vector pBICRNAi3 for silencing CfTOM1 was constructed and transformed to pepper cultivar Tiandeng No. 1 by Agrobacterium tumefaciens-infection. Thirty three selection marker-positive regenerated lines were detained.To lines and offsprings of T_1 and T_2 lines were further confirmed by PCR Sequencing and Southern analysis of the transgene to select the transgenic lines.T_2 lines were assayed for resistance to TMV , cucumber mosaic virus (CMV) and tomato mosaic virus (ToMV). It turned out that the transgenic pepper expressed significantly enhanced disease resistance to these three viruses. The results suggest that CfTOM1 is a host factor gene possibly involved in the replication of virus in pepper, likely throught the similar mechanism as AtTOM1 in Arabidopsis.
引文
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