Department of Agricultural Biotechnology, College of Agriculture & Life Science, Seoul National University, Seoul, Republic of Korea.
Research Institute for Agriculture and Life Sciences, Seoul National University, Seoul, Republic of Korea.
Pest Manag Sci. 2020 May;76(5):1699-1704. doi: 10.1002/ps.5692. Epub 2019 Dec 17.
Sacbrood virus (SBV) is a fatal viruses that infects the Asian honey bee, Apis cerana in Korea. Recently, RNA interference (RNAi) has been suggested as a promising strategy for the suppression of honey bee viruses. For the efficient control of SBV infection using RNAi, simple and cost-effective methods to produce double-stranded RNA (dsRNA) are needed.
To develop a dsRNA production platform using Bacillus thuringiensis (Bt), pBTdsSBV-VP1 vector was constructed in which the SBV vp1 gene was located between two oppositely oriented cyt promoters. Both strands of the vp1 gene were bidirectionally transcribed under the control of the sporulation-dependent cyt promoter in Bt cells transformed with pBTdsSBV-VP1, and the resulting dsRNA was easily extracted from the Bt transformant, Bt 4Q7/pBTdsSBV-VP1, by inducing its autolysis. The replication of SBV was dramatically suppressed in A. cerana workers who ingested the dsRNA produced from the Bt 4Q7/pBTdsSBV-VP1.
In this study, we successfully silenced SBV in its host, A. cerana, by the application of exogenous dsRNA produced from an entomopathogenic bacteria, Bt. These results suggested that Bt could be a useful dsRNA production platform to control viral pathogens in their host insects. © 2019 Society of Chemical Industry.
Sacbrood 病毒（SBV）是一种致命的病毒，感染韩国的亚洲蜜蜂（Apis cerana）。最近，RNA 干扰（RNAi）被认为是抑制蜜蜂病毒的一种有前途的策略。为了利用 RNAi 有效控制 SBV 感染，需要简单且具有成本效益的双链 RNA（dsRNA）生产方法。
为了利用苏云金芽孢杆菌（Bt）开发 dsRNA 生产平台，构建了 pBTdsSBV-VP1 载体，其中 SBV vp1 基因位于两个反向 cyt 启动子之间。在转化有 pBTdsSBV-VP1 的 Bt 细胞中，vp1 基因的两条链在依赖于孢子形成的 cyt 启动子的控制下双向转录，并且源自 Bt 转化体（Bt 4Q7/pBTdsSBV-VP1）的 dsRNA 可通过诱导其自溶轻松提取。摄入来自 Bt 4Q7/pBTdsSBV-VP1 的 dsRNA 的 A. cerana 工蜂中 SBV 的复制受到显著抑制。
在这项研究中，我们通过应用来自昆虫病原细菌 Bt 的外源性 dsRNA 成功地沉默了其宿主 A. cerana 中的 SBV。这些结果表明，Bt 可以成为一种有用的 dsRNA 生产平台，用于控制宿主昆虫中的病毒病原体。© 2019 化学工业学会。
