State Key Laboratory of Biocatalysis and Enzyme Engineering, Hubei Hongshan Laboratory, School of Life Sciences, Hubei University, Wuhan, China.
Shenzhen Branch, Guangdong Laboratory of Lingnan Modern Agriculture, Key Laboratory of Synthetic Biology, Ministry of Agriculture and Rural Affairs, Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Shenzhen, China.
Plant Biotechnol J. 2024 Jul;22(7):2010-2019. doi: 10.1111/pbi.14321. Epub 2024 Mar 1.
RNA interference (RNAi) has emerged as an efficient technology for pest control by silencing the essential genes of targeted insects. Owing to its nucleotide sequence-guided working mechanism, RNAi has a high degree of species-specificity without impacts on non-target organisms. However, as plants are inevitably under threat by two or more insect pests in nature, the species-specific mode of RNAi-based technology restricts its wide application for pest control. In this study, we artificially designed an intermediate dsRNA (iACT) targeting two β-Actin (ACT) genes of sap-sucking pests Bemisia tabaci and Myzus persicae by mutual correction of their mismatches. When expressing hairpin iACT (hpiACT) from tobacco nuclear genome, transgenic plants are well protected from both B. tabaci and M. persicae, either individually or simultaneously, as evidenced by reduced fecundity and suppressed ACT gene expression, whereas expression of hpRNA targeting BtACT or MpACT in transgenic tobacco plants could only confer specific resistance to either B. tabaci or M. persicae, respectively. In sum, our data provide a novel proof-of-concept that two different insect species could be simultaneously controlled by artificial synthesis of dsRNA with sequence optimization, which expands the range of transgenic RNAi methods for crop protection.
RNA 干扰 (RNAi) 通过沉默靶标昆虫的必需基因,已成为一种有效的害虫防治技术。由于其核苷酸序列指导的工作机制,RNAi 具有高度的物种特异性,而不会对非靶标生物产生影响。然而,由于植物在自然界中不可避免地受到两种或更多种害虫的威胁,基于 RNAi 的技术的物种特异性模式限制了其在害虫防治中的广泛应用。在本研究中,我们通过相互校正其错配,人工设计了一种针对刺吸式害虫烟粉虱和桃蚜的两个β-肌动蛋白 (ACT) 基因的中间 dsRNA (iACT)。当从烟草核基因组中表达发夹 iACT (hpiACT) 时,转基因植物可免受烟粉虱和桃蚜的侵害,无论是单独还是同时,这表现为生育力降低和 ACT 基因表达受到抑制,而靶向 BtACT 或 MpACT 的 hpRNA 在转基因烟草植物中的表达只能分别赋予对烟粉虱或桃蚜的特异性抗性。总之,我们的数据提供了一个新的概念验证,即通过序列优化的人工合成 dsRNA 可以同时控制两种不同的昆虫物种,这扩展了转基因 RNAi 方法在作物保护中的应用范围。