Dandan Feng , Huizhen Guo , Jie Li , Jiayi Liu , Dan Zhao , Hui Zhao , Xinyi He , Qingyou Xia , Liang Jiang
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In BmE cells, knockout of <em>BmWh2</em> significantly increased viral DNA content, while overexpression of <em>BmWh2</em> had the opposite effects, indicating that <em>BmWh2</em> could inhibit BmNPV proliferation. A transgenic vector overexpressing <em>BmWh2</em> was constructed under the control of the A4 promoter, and two transgenic silkworm lines A4Wh2–1/2 were generated <em>via</em> embryo microinjection. qPCR analysis confirmed that the expression level of <em>BmWh2</em> in A4Wh2 silkworms was significantly increased. After oral infection with BmNPV, the mortality of A4Wh2–2 was 24 % lower than that of non-transgenic WT, and the viral DNA content in A4Wh2–2 was 8 % of that in WT. The mRNA levels of <em>ATG6</em> and <em>ATG8</em> were increased in both BmE cells overexpressing <em>BmWh2</em> and A4Wh2 silkworms, implying that <em>BmWh2</em> inhibited viral proliferation by increasing <em>ATG</em> expression. This study is the first report of an ABC transporter involved in antiviral immunity in <em>B. mori</em>, providing a theoretical reference for pesticide resistance.</div></div>","PeriodicalId":19828,"journal":{"name":"Pesticide Biochemistry and Physiology","volume":"215 ","pages":"Article 106704"},"PeriodicalIF":4.0000,"publicationDate":"2025-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"ABC transporter BmWh2 enhances antiviral defense against baculovirus in silkworm via ATG upregulation\",\"authors\":\"Dandan Feng , Huizhen Guo , Jie Li , Jiayi Liu , Dan Zhao , Hui Zhao , Xinyi He , Qingyou Xia , Liang Jiang\",\"doi\":\"10.1016/j.pestbp.2025.106704\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><div>Baculoviruses are widely used as biological pesticides for pest control. <em>Bombyx mori</em> nucleopolyhedrovirus (BmNPV) is a typical baculovirus that causes serious economic losses in the sericulture. ATP-binding cassette (ABC) transporters are widely distributed in various organisms, from viruses to humans, and play important roles in multiple biological processes. However, their roles in the interactions between viruses and insect hosts remain largely unknown. In this study, we used molecular cloning, reverse transcription-PCR, and quantitative PCR (qPCR) to investigate the function of the silkworm ABC transporter <em>BmWh2</em> during BmNPV infection. The results showed that <em>BmWh2</em> was upregulated after BmNPV infection. In BmE cells, knockout of <em>BmWh2</em> significantly increased viral DNA content, while overexpression of <em>BmWh2</em> had the opposite effects, indicating that <em>BmWh2</em> could inhibit BmNPV proliferation. A transgenic vector overexpressing <em>BmWh2</em> was constructed under the control of the A4 promoter, and two transgenic silkworm lines A4Wh2–1/2 were generated <em>via</em> embryo microinjection. qPCR analysis confirmed that the expression level of <em>BmWh2</em> in A4Wh2 silkworms was significantly increased. After oral infection with BmNPV, the mortality of A4Wh2–2 was 24 % lower than that of non-transgenic WT, and the viral DNA content in A4Wh2–2 was 8 % of that in WT. The mRNA levels of <em>ATG6</em> and <em>ATG8</em> were increased in both BmE cells overexpressing <em>BmWh2</em> and A4Wh2 silkworms, implying that <em>BmWh2</em> inhibited viral proliferation by increasing <em>ATG</em> expression. This study is the first report of an ABC transporter involved in antiviral immunity in <em>B. mori</em>, providing a theoretical reference for pesticide resistance.</div></div>\",\"PeriodicalId\":19828,\"journal\":{\"name\":\"Pesticide Biochemistry and Physiology\",\"volume\":\"215 \",\"pages\":\"Article 106704\"},\"PeriodicalIF\":4.0000,\"publicationDate\":\"2025-09-19\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Pesticide Biochemistry and Physiology\",\"FirstCategoryId\":\"97\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S0048357525004171\",\"RegionNum\":1,\"RegionCategory\":\"农林科学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"BIOCHEMISTRY & MOLECULAR BIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Pesticide Biochemistry and Physiology","FirstCategoryId":"97","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0048357525004171","RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
引用次数: 0
摘要
杆状病毒是一种广泛应用于害虫防治的生物农药。家蚕核多角体病毒(Bombyx mori nuclear polyhedrovirus, BmNPV)是一种典型的杆状病毒,给蚕桑养殖业造成严重的经济损失。atp结合盒(ABC)转运体广泛分布于从病毒到人类的各种生物体中,并在多种生物过程中发挥重要作用。然而,它们在病毒和昆虫宿主之间相互作用中的作用在很大程度上仍然未知。本研究采用分子克隆、反转录PCR和定量PCR (qPCR)技术,研究了家蚕ABC转运蛋白BmWh2在BmNPV感染过程中的功能。结果表明,BmWh2在BmNPV感染后表达上调。在BmE细胞中,敲除BmWh2可显著增加病毒DNA含量,而过表达BmWh2则相反,表明BmWh2可抑制BmNPV的增殖。在A4启动子的控制下构建了过表达BmWh2的转基因载体,并通过胚胎显微注射获得了2个转基因家蚕品系A4Wh2-1/2。qPCR分析证实,BmWh2在A4Wh2家蚕中的表达水平显著升高。经口腔感染BmNPV后,A4Wh2 - 2的死亡率比非转基因WT低24%,A4Wh2 - 2的病毒DNA含量为非转基因WT的8%。过表达BmWh2和A4Wh2的家蚕BmE细胞中ATG6和ATG8的mRNA水平均升高,表明BmWh2通过提高ATG的表达抑制病毒增殖。本研究首次报道了ABC转运体参与家蚕抗病毒免疫的研究,为家蚕的农药抗性研究提供了理论依据。
ABC transporter BmWh2 enhances antiviral defense against baculovirus in silkworm via ATG upregulation
Baculoviruses are widely used as biological pesticides for pest control. Bombyx mori nucleopolyhedrovirus (BmNPV) is a typical baculovirus that causes serious economic losses in the sericulture. ATP-binding cassette (ABC) transporters are widely distributed in various organisms, from viruses to humans, and play important roles in multiple biological processes. However, their roles in the interactions between viruses and insect hosts remain largely unknown. In this study, we used molecular cloning, reverse transcription-PCR, and quantitative PCR (qPCR) to investigate the function of the silkworm ABC transporter BmWh2 during BmNPV infection. The results showed that BmWh2 was upregulated after BmNPV infection. In BmE cells, knockout of BmWh2 significantly increased viral DNA content, while overexpression of BmWh2 had the opposite effects, indicating that BmWh2 could inhibit BmNPV proliferation. A transgenic vector overexpressing BmWh2 was constructed under the control of the A4 promoter, and two transgenic silkworm lines A4Wh2–1/2 were generated via embryo microinjection. qPCR analysis confirmed that the expression level of BmWh2 in A4Wh2 silkworms was significantly increased. After oral infection with BmNPV, the mortality of A4Wh2–2 was 24 % lower than that of non-transgenic WT, and the viral DNA content in A4Wh2–2 was 8 % of that in WT. The mRNA levels of ATG6 and ATG8 were increased in both BmE cells overexpressing BmWh2 and A4Wh2 silkworms, implying that BmWh2 inhibited viral proliferation by increasing ATG expression. This study is the first report of an ABC transporter involved in antiviral immunity in B. mori, providing a theoretical reference for pesticide resistance.
期刊介绍:
Pesticide Biochemistry and Physiology publishes original scientific articles pertaining to the mode of action of plant protection agents such as insecticides, fungicides, herbicides, and similar compounds, including nonlethal pest control agents, biosynthesis of pheromones, hormones, and plant resistance agents. Manuscripts may include a biochemical, physiological, or molecular study for an understanding of comparative toxicology or selective toxicity of both target and nontarget organisms. Particular interest will be given to studies on the molecular biology of pest control, toxicology, and pesticide resistance.
Research Areas Emphasized Include the Biochemistry and Physiology of:
• Comparative toxicity
• Mode of action
• Pathophysiology
• Plant growth regulators
• Resistance
• Other effects of pesticides on both parasites and hosts.