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温室喷施 dsRNA 防治辣椒上的西花蓟马,Frankliniella occidentalis。

Greenhouse test of spraying dsRNA to control the western flower thrips, Frankliniella occidentalis, infesting hot peppers.

机构信息

Department of Plant Medicals, College of Life Sciences, Andong National University, Andong, 36729, Korea.

Genolution, Inc, Seoul, 05836, Korea.

出版信息

BMC Biotechnol. 2023 Apr 4;23(1):10. doi: 10.1186/s12896-023-00780-y.

Abstract

BACKGROUND

The western flower thrips Frankliniella occidentalis is an insect pest that damages various crops, including hot peppers. It is a vector of a plant pathogen, tomato spotted wilt virus. To control this pest, chemical insecticides have been used in the past, but the control efficacy is unsatisfactory owing to rapid resistance development by F. occidentalis.

METHODOLOGY

This study reports a novel control technology against this insect pest using RNA interference (RNAi) of the vacuolar-type ATPase (vATPase) expression. Eight subunit genes (vATPase-A ∼ vATPase-H) of vATPase were obtained from the F. occidentalis genome and confirmed for their expressions at all developmental stages.

RESULTS

Double-stranded RNAs (dsRNAs) specific to the eight subunit genes were fed to larvae and adults, which significantly suppressed the corresponding gene expressions after 24-h feeding treatment. These RNAi treatments resulted in significant mortalities, in which the dsRNA treatments at ∼2,000 ppm specific to vATPase-A or vATPase-B allowed complete control efficacy near 100% mortality in 7 days after treatment. To prevent dsRNA degradation by the digestive proteases during oral feeding, dsRNAs were formulated in a liposome and led to an enhanced mortality of the larvae and adults of F. occidentalis. The dsRNAs were then sprayed at 2,000 ppm on F. occidentalis infesting hot peppers in a greenhouse, which resulted in 53.5-55.9% control efficacy in 7 days after treatment. Even though the vATPases are conserved in different organisms, the dsRNA treatment was relatively safe for non-target insects owing to the presence of mismatch sequences compared to the dsRNA region of F. occidentalis.

CONCLUSION

These results demonstrate the practical feasibility of spraying dsRNA to control F. occidentalis infesting crops.

摘要

背景

西部花蓟马(Frankliniella occidentalis)是一种破坏包括辣椒在内的各种作物的害虫,它还是植物病原体番茄斑萎病毒的载体。为了控制这种害虫,过去曾使用化学杀虫剂,但由于西方花蓟马快速产生抗药性,控制效果并不理想。

方法

本研究报告了一种利用液泡型 ATP 酶(vATPase)表达的 RNA 干扰(RNAi)来控制这种昆虫的新技术。从西方花蓟马基因组中获得了 vATPase 的 8 个亚基基因(vATPase-A~vATPase-H),并在所有发育阶段都证实了它们的表达。

结果

dsRNA 特异性针对这 8 个亚基基因,喂食给幼虫和成虫,喂食 24 小时后,dsRNA 显著抑制了相应基因的表达。这些 RNAi 处理导致了显著的死亡率,vATPase-A 或 vATPase-B 的 dsRNA 处理浓度约为 2000ppm 时,处理后 7 天内死亡率接近 100%。为了防止在口服喂食时 dsRNA 被消化蛋白酶降解,dsRNA 被包裹在脂质体中,导致西方花蓟马幼虫和成虫的死亡率显著提高。然后将 2000ppm 的 dsRNA 溶液喷洒在温室中受西方花蓟马侵害的辣椒上,处理后 7 天内的防治效果为 53.5-55.9%。尽管不同生物体中的 vATPases 是保守的,但与西方花蓟马的 dsRNA 区域相比,由于存在错配序列,dsRNA 处理对非靶标昆虫相对安全。

结论

这些结果表明,喷洒 dsRNA 来控制侵害作物的西方花蓟马是可行的。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f7b0/10074877/0321c7949b5b/12896_2023_780_Fig1_HTML.jpg

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