中文摘要：西瓜是一种在生长过程中遭受各种病害的一年生蔬菜作物。基因工程是提高植物抗病性的有效工具。本文为母本西瓜品种研发了一种有效的农杆菌介导转化方法。利用该方法，重新获得了17种可能表达人工microRNAs（以CMV 2a/2b基因为靶标）的转基因西瓜，而由于转基因西瓜自花授粉的技术难度，其中只有7个成功收获了T1代种子。PCR和Southern blot分析发现，转基因片段已被成功导入和整合到其中3种转基因西瓜的基因组中。DAS-ELISA分析表明，在非转基因品种中可检测到CMV，但在转基因品种中检不到CMV，可见，转基因品种可抵抗CMV侵染。
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
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