中文摘要：西瓜是一年生作物，生长过程中会遭受各种病害的影响。基因工程是提高植物抗病性的有效工具。本文提出了一种有效的农杆菌介导母本西瓜系的转化设计方案。采用这种转化方法，可以获得17个假定的表达人工小RNAs（以CMV2a/2b基因为靶标）的转基因植株，由于转基因西瓜自花授粉的技术难点，其中只有7个转基因植株成功获得了T1代种子。PCR和Southern杂交分析表明，这7个转基因植株中有3个转基因植株的基因组中成功导入并整合了转移片段。DAS-ELISA分析表明，可在非转基因株系中检测到CMV，但在转基因株系中检测不到，转基因株系表现出对CMV侵染的抗性。
外文摘要：Watermelon is an annual vegetable crop that suffers from various diseases during growth. Genetic engineering is an effective tool for improving plant disease resistance. This paper presents an effective transformation protocol designed for a female parent watermelon line mediated by Agrobacterium. In this protocol, 2 times MS Fe-EDTA were supplemented into Murashige and Skoog (MS) medium to suppress the chlorosis during watermelon explants culturing, and the optimal preculture time (5 days) and coculture time (5 days) were screened out to improve the frequency of shoots regeneration. Using our protocol, 17 putative transgenic plants expressing artificial microRNAs that target Cucumber mosaic virus (CMV) 2a/2b genes were recovered, and seven of them were succeeded in obtaining T1 generation seeds because of the technique difficulties of self-pollination of the transgenic watermelon lines. PCR and Southern blot analyses confirmed that the transferred fragment was successfully imported and integrated into the genome of three of these seven transgenic lines. DAS-ELISA assay indicated that the CMV can be detected in the nontransgenic line but not in the transgenic lines, and the transgenic lines displayed resistance to CMV infection.
外文关键词：Watermelon；Transformation；Cucumber mosaic virus；Resistance；Artificial microRNA
作者：Liu, Lifeng；Gu, Qinsheng；Ijaz, Raina；等
作者单位：华中农业大学
期刊名称：SCIENTIA HORTICULTURAE
期刊影响因子：1.538
出版年份：2016
出版刊次：6
点击下载：通过农杆菌介导转化法可生产抗黄瓜花叶病毒的转基因西瓜