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首页> 美国卫生研究院文献>Springer Open Choice >Construction of Corynebacterium glutamicum cells as containers encapsulating dsRNA overexpressed for agricultural pest control
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Construction of Corynebacterium glutamicum cells as containers encapsulating dsRNA overexpressed for agricultural pest control

机译：谷氨酸棒杆菌细胞构建超表达用于农业病虫害防治的dsRNA容器的构建

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Double-stranded RNA (dsRNA) inducing RNA interference (RNAi) is expected to be applicable to management of agricultural pests. In this study, we selected a ladybird beetle, Henosepilachna vigintioctopunctata, as a model target pest that devours vegetable leaves, and examined the effects of feeding the pest sterilized microbes highly accumulating a target dsRNA for RNAi induction. We constructed an efficient production system for diap1*-dsRNA, which suppresses expression of the essential gene diap1 (encoding death-associated inhibitor of apoptosis protein 1) in H. vigintioctopunctata, using an industrial strain of Corynebacterium glutamicum as the host microbe. The diap1*-dsRNA was overproduced in C. glutamicum by convergent transcription using a strong promoter derived from corynephage BFK20, and the amount of dsRNA accumulated in fermented cells reached about 75 mg per liter of culture. In addition, we developed a convenient method for stabilizing the dsRNA within the microbes by simply sterilizing the diap1*-dsRNA-expressing C. glutamicum cells with ethanol. When the sterilized microbes containing diap1*-dsRNA were fed to larvae of H. vigintioctopunctata, diap1 expression in the pest was suppressed, and the leaf-feeding activity of the larvae declined. These results suggest that this system is capable of producing stabilized dsRNA for RNAi at low cost, and it will open a way to practical application of dsRNA as an environmentally-friendly agricultural insecticide.Electronic supplementary materialThe online version of this article (10.1007/s00253-019-10113-9) contains supplementary material, which is available to authorized users.
机译：诱导RNA干扰（RNAi）的双链RNA（dsRNA）有望应用于农业害虫的治理。在这项研究中，我们选择了瓢虫甲虫Henosepilachna vigintioctopunctata作为吞食蔬菜叶片的标准目标害虫，并研究了喂养高度积累了目标dsRNA的有害生物无菌微生物对RNAi诱导的影响。我们构建了diap1 * -dsRNA的高效生产系统，该系统使用谷氨酸棒杆菌的工业菌株作为宿主微生物来抑制必需基因diap1（编码死亡相关凋亡蛋白1的死亡抑制剂）的表达。 diap1 * -dsRNA在谷氨酸棒杆菌中通过使用源自棒状噬菌体BFK20的强启动子会聚转录而过量产生，并且在发酵细胞中积累的dsRNA量达到每升培养液约75 mg。此外，我们开发了一种简便的方法，可通过简单地用乙醇对表达diap1 * -dsRNA的谷氨酸棒状杆菌细胞进行灭菌来稳定微生物中的dsRNA。当将含有diap1 * -dsRNA的无菌微生物饲喂到葡萄球菌的幼虫时，害虫中diap1的表达受到抑制，幼虫的叶片摄食活性下降。这些结果表明，该系统能够以低成本生产用于RNAi的稳定化dsRNA，它将为将dsRNA用作环境友好型农业杀虫剂开辟一条途径。电子补充材料本文的在线版本（10.1007 / s00253- 019-10113-9）包含补充材料，授权用户可以使用。

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