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Orally delivered dsRNA induces knockdown of target genes and mortality in the Asian long-horned beetle, Anoplophora glabripennis.
Archives of Insect Biochemistry and Physiology ( IF 1.5 ) Pub Date : 2020-04-15 , DOI: 10.1002/arch.21679
Ramesh Kumar Dhandapani 1 , Jian Jun Duan 2 , Subba Reddy Palli 1
Affiliation  

The Asian long‐horned beetle (ALB) Anoplophora glabripennis is a serious invasive forest pest in several countries, including the United States. Methods available to manage or eradicate this pest are extremely limited, but RNA interference (RNAi) technology is a potentially effective method to control ALB. In this study, we used sucrose feeding bioassay for oral delivery of double‐strand RNA (dsRNA) to ALB larvae. 32P‐labeled dsRNA orally delivered to ALB larvae using the sucrose droplet feeding method was processed to small interfering RNA. Feeding neonate larvae with dsRNA targeting genes coding for the inhibitor of apoptosis (IAP), vacuolar sorting protein SNF7 (SNF7), and snakeskin (SSK) induced knockdown of target genes and mortality. Feeding 2 µg of dsRNA per day for 3 days did not induce a significant decrease in the expression of target genes or mortality. However, feeding 5 or 10 µg of dsRNA per day for 3 days induced a significant decrease in the expression of target genes and 50–90% mortality. Interestingly, feeding 2.5 µg each of dsIAP plus dsSNF7, dsIAP plus dsSSK, or dsSNF7 plus dsSSK per day for 3 days induced a significant decrease in the expression of both target genes and approximately 80% mortality. Our findings demonstrate that orally delivered dsRNA induces target gene knockdown and mortality in ALB neonate larvae and RNAi technology may have the potential for effective ALB control.

中文翻译:

口服递送的dsRNA诱导了亚洲长角甲虫Anoplophora glabripennis的靶基因敲低和死亡率。

亚洲长角甲虫(Anoplophora glabripennis)是包括美国在内的多个国家的严重入侵森林害虫。处理或根除这种有害生物的方法极为有限,但是RNA干扰(RNAi)技术是控制ALB的潜在有效方法。在这项研究中,我们使用蔗糖喂养生物测定法将双链RNA(dsRNA)口服递送至ALB幼虫。使用蔗糖液滴饲喂法将32 P标记的dsRNA口服递送至ALB幼虫,然后处理成小的干扰RNA。用编码凋亡抑制剂(IAP),液泡分选蛋白SNF7(SNF7)和蛇皮(SSK)的dsRNA靶向基因喂养新生幼虫。)诱导的靶基因敲低和死亡率。每天饲喂2 µg dsRNA,持续3天,并不会引起靶基因表达或死亡率的显着降低。但是,每天饲喂5或10 µg dsRNA连续3天,导致靶基因表达显着下降,死亡率降低50-90%。有趣的是,每天分别饲喂2.5 µg dsIAP和dsSNF7,dsIAP和dsSSK或dsSNF7和dsSSK,持续3天,这两个靶基因的表达均显着下降,死亡率约80%。我们的发现表明,口服递送的dsRNA可以诱导靶基因敲低,并且在ALB新生幼虫中致死,而RNAi技术可能具有有效控制ALB的潜力。
更新日期:2020-04-15
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