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通过在番茄叶片上局部施用双链RNA对v-ATPase B和保幼激素结合蛋白基因进行RNA干扰抑制:开发用于防治南美番茄潜麦蛾(鳞翅目:麦蛾科)的生物农药

RNA Interference Suppression of v-ATPase B and Juvenile Hormone Binding Protein Genes Through Topically Applied dsRNA on Tomato Leaves: Developing Biopesticides to Control the South American Pinworm, (Lepidoptera: Gelechiidae).

作者信息

Ramkumar Govindaraju, Asokan Ramasamy, Prasannakumar N R, Kariyanna B, Karthi Sengodan, Alwahibi Mona S, Elshikh Mohamed Soliman, Abdel-Megeed Ahmed, Ghaith Aml, Senthil-Nathan Sengottayan, Kalaivani Kandaswamy, Hunter Wayne Brian, Krutmuang Patcharin

机构信息

Division of Biotechnology, ICAR-Indian Institute of Horticultural Research (IIHR), Bengaluru, India.

Division of Entomology and Nematology, ICAR-Indian Institute of Horticultural Research (IIHR), Bengaluru, India.

出版信息

Front Physiol. 2021 Nov 18;12:742871. doi: 10.3389/fphys.2021.742871. eCollection 2021.

DOI:10.3389/fphys.2021.742871
PMID:34867448
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8637209/
Abstract

The South American pinworm (Meyrick) (Family: Gelechiidae) is one of the most devastating lepidopteran pests in the developing countries of South America, Africa, and Asia. This pest is classified as the most serious threat for tomato production worldwide. In the present study, we analyzed RNAi-mediated control through exogenously applied dsRNA delivery on tomato. The dsRNA treatments were made to target the juvenile hormone binding protein and the v-ATPase B. Both mRNA targets were cloned, validated by sequencing, and used to produce each dsRNA. After treatments the relative transcript expression was analyzed using qRTPCR to assess to efficacy of RNAi. A leaf-dip assay was used to provide late 2nd instar larvae three feeding access periods: 24, 48, and 72 h, to evaluate the effect of gene silencing of each target. Larvae were fed tomato leaves coated with five different RNAi concentrations (10, 20, 30, 40, and 50 micrograms/centimeter-squared), that suppressed two genes (juvenile hormone protein, JHBP, and vacuolar-type adenosine triphosphatase enzyme, v-ATPase). Treatments with dsRNA showed a significant increase in mortality at 24, 48, and 72 h after ingestion ( < 0.01, α = 0.05), along with reduced leaf damage, and increased feeding deterrence. The results suggest that these two RNAi products may provide a suitable treatment for control of this and other lepidopteran pests.

摘要

南美番茄潜叶蛾(梅里克)(麦蛾科)是南美洲、非洲和亚洲发展中国家最具毁灭性的鳞翅目害虫之一。这种害虫被列为全球番茄生产面临的最严重威胁。在本研究中,我们通过向番茄外源施用双链RNA(dsRNA)来分析RNA干扰介导的防治效果。dsRNA处理针对保幼激素结合蛋白和液泡型ATP酶B。两个mRNA靶标均被克隆、测序验证,并用于生产各自的dsRNA。处理后,使用定量逆转录聚合酶链反应(qRTPCR)分析相对转录本表达,以评估RNA干扰的效果。采用浸叶试验为二龄后期幼虫提供24小时、48小时和72小时三个取食时段,以评估每个靶标的基因沉默效果。用涂有五种不同RNA干扰浓度(10、20、30、40和50微克/平方厘米)的番茄叶喂养幼虫,这些浓度可抑制两个基因(保幼激素蛋白JHBP和液泡型腺苷三磷酸酶v-ATPase)。dsRNA处理在幼虫摄食后24小时、48小时和72小时导致死亡率显著增加(<0.01,α=0.05),同时叶片损伤减少,取食抑制作用增强。结果表明,这两种RNA干扰产物可能为防治这种及其他鳞翅目害虫提供合适的处理方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3fee/8637209/a8569274b7a9/fphys-12-742871-g0005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3fee/8637209/6a34ed9b0dd1/fphys-12-742871-g0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3fee/8637209/3bfb1c018e53/fphys-12-742871-g0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3fee/8637209/13119609aff6/fphys-12-742871-g0003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3fee/8637209/d27a4bfdab7c/fphys-12-742871-g0004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3fee/8637209/a8569274b7a9/fphys-12-742871-g0005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3fee/8637209/6a34ed9b0dd1/fphys-12-742871-g0001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3fee/8637209/3bfb1c018e53/fphys-12-742871-g0002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3fee/8637209/13119609aff6/fphys-12-742871-g0003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3fee/8637209/d27a4bfdab7c/fphys-12-742871-g0004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3fee/8637209/a8569274b7a9/fphys-12-742871-g0005.jpg

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多化性斜纹夜蛾幼虫和成虫中与氯菊酯抗性相关的细胞色素 P450 和酯酶基因的过度表达。
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