Efficient silencing gene construct for resistance to multiple common bean (Phaseolus vulgaris L.) viruses
Efficient silencing gene construct for resistance to multiple common bean (Phaseolus vulgaris L.) viruses
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DOI：
10.1007/s13205-020-02276-4
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发表时间：
2020-05-30
期刊：
影响因子：
2.8
通讯作者：
Shams-bakhsh, Masoud
中科院分区：
文献类型：
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作者：
Azizi, Abdolbaset;Verchot, Jeanmarie;Shams-bakhsh, Masoud
一种有希望的策略是利用RNA沉默结构来转化植物以抵抗多种病原体，从而使植物对植物产生抗病作用。菜豆受到菜豆普通花叶病毒(BCMV)、菜豆普通花叶坏死病毒(BCMNV)和黄瓜花叶病毒(CMV)的严重危害。在本研究中，我们制备了产生正义、反义和发夹RNA(HpRNA)结构的载体，以靶向单个和多个病毒。利用农杆菌(GV3101)在烟草和菜豆植株中的瞬时表达分析了这些构建物的沉默效率。结果表明，该方法显著减少了白粉病的症状和病毒的积累。一般来说，所制备的构建物的效率分别为发夹、反义和正义，并且单基因和多基因构建物在减少病毒积累方面存在显著差异，多基因构建体显示出较高的效果。本研究表明，利用含有秀丽线虫基因Ced-9或植物基因AtBag-4的双元载体农杆菌与含有BCMV-CP、BCMNV-HC-Pro和CMV-2b多个基因片段的农杆菌pFGC-BNC.h混合悬浮液，提高了pFGC-BNC.h的转化效率。我们在这些转基因豆类计划中显示了病毒积累的减少。
One promising strategy to engineer plants that are resistant to plant pathogens involves transforming plants with RNA silencing constructs for resistance to multiple pathogens. Garden bean is significantly damaged by bean common mosaic virus (BCMV), bean common mosaic necrosis virus (BCMNV) and cucumber mosaic virus (CMV). In this study, we prepared constructs producing sense, antisense and hairpin RNA (hpRNA) structures to target single as well as multiple viruses. Silencing efficiency of these constructions was analyzed using Agrobacterium (GV3101) transient expression in Nicothinia bethamiana and Phaseolus vulgaris plants. The results showed significantly reduced disease symptoms and virus accumulation in N. bethamiana plants. Generally, the efficiency of the prepared constructs was hairpin, antisense and sense, respectively, and also, there was a significant difference between mono-gene and multiple-gene constructs for reducng virus accumulation and the multiple-gene constructs showed higher effectiveness. Experiments in this study showed that using Agrobacterium harboring binary constructs containing a Caenorhabditis elegans gene, Ced-9, or a plant gene, AtBag-4, anti-apoptosis gene as a mix suspension with an Agrobacterium containing pFGC-BNC.h, a plasmid containing multiple gene fragments consisting of BCMV-CP, BCMNV-HC-Pro and CMV-2b, improved the efficiency of pFGC-BNC.h transformation. We showed reduced virus accumulation in these transgenic bean plans.