A cucumber green mottle mosaic virus vector for virus-induced gene silencing in cucurbit plants
A cucumber green mottle mosaic virus vector for virus-induced gene silencing in cucurbit plants
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DOI：
10.1186/s13007-020-0560-3
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发表时间：
2020-02-03
期刊：
影响因子：
5.1
通讯作者：
Gu, Qinsheng
中科院分区：
文献类型：
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作者：
Liu, Mei;Liang, Zhiling;Gu, Qinsheng
背景 葫芦科植物生产的水果或蔬菜在全世界范围内具有重要的饮食重要性和经济意义。已发表的至少 11 个葫芦科物种的基因组正在推动基因挖掘和新的育种策略，但由于转化效率低，葫芦科遗传转化作为基因功能验证的工具是不切实际的。病毒诱导的基因沉默（VIGS）是一种潜在的替代工具。到目前为止，可用于葫芦科植物的理想 VIGS 载体还很少。结果在这里，我们描述了一种源自黄瓜绿斑驳花叶病毒（CGMMV）的新 VIGS 载体，CGMMV 是一种自然感染葫芦科植物的单部分病毒。我们表明，CGMMV 载体能够在模型植物本塞姆氏烟草和葫芦科植物（包括西瓜、甜瓜、黄瓜和葫芦）中诱导八氢番茄红素去饱和酶（PDS）基因的有效沉默。用具有正义导向或发夹cDNA形式的长度为69-300bp的PDS序列的CGMMV载体感染，通过PDS沉默导致本塞姆氏烟草和葫芦科植物的光漂白表型。其他结果表明，PDS 基因的沉默可以持续两个月以上，并且基于 CGMMV 的载体的沉默效果可以被传代。结论 这些结果表明，CGMMV 载体可以作为一种强大且易于使用的工具，用于表征基因功能、控制病毒病原体，甚至在葫芦科植物中进行抗性育种。此外，本研究对于开发不同的病毒载体具有相当重要的参考价值。
Background Cucurbits produce fruits or vegetables that have great dietary importance and economic significance worldwide. The published genomes of at least 11 cucurbit species are boosting gene mining and novel breeding strategies, however genetic transformation in cucurbits is impractical as a tool for gene function validation due to low transformation efficiency. Virus-induced gene silencing (VIGS) is a potential alternative tool. So far, very few ideal VIGS vectors are available for cucurbits. Results Here, we describe a new VIGS vector derived from cucumber green mottle mosaic virus (CGMMV), a monopartite virus that infects cucurbits naturally. We show that the CGMMV vector is competent to induce efficient silencing of the phytoene desaturase (PDS) gene in the model plant Nicotiana benthamiana and in cucurbits, including watermelon, melon, cucumber and bottle gourd. Infection with the CGMMV vector harboring PDS sequences of 69-300 bp in length in the form of sense-oriented or hairpin cDNAs resulted in photobleaching phenotypes in N. benthamiana and cucurbits by PDS silencing. Additional results reflect that silencing of the PDS gene could persist for over two months and the silencing effect of CGMMV-based vectors could be passaged. Conclusions These results demonstrate that CGMMV vector could serve as a powerful and easy-to-use tool for characterizing gene function, controlling viral pathogens or even performing resistance breeding in cucurbits. Moreover, this study will possess considerable important reference value for developing different viral vectors.