Lin Li, Xinru Pang, Chenyang Wang, Yihua Yang, Yidong Wu
{"title":"表达四泛素基因 HaTSPAN1 的 T92C 等位基因的基于 piggyBac 的转基因 Helicoverpa armigera 对苏云金芽孢杆菌毒素 Cry1Ac 具有显性抗性","authors":"Lin Li, Xinru Pang, Chenyang Wang, Yihua Yang, Yidong Wu","doi":"10.1016/j.pestbp.2024.106096","DOIUrl":null,"url":null,"abstract":"<div><p>Transgenic crops producing insecticidal proteins from <em>Bacillus thuringiensis</em> (Bt) have revolutionized pest control. However, the evolution of resistance by target pests poses a significant threat to the long-term success of Bt crops. Understanding the genetics and mechanisms underlying Bt resistance is crucial for developing resistance detection methods and management tactics. The T92C mutation in a tetraspanin gene (<em>HaTSPAN1</em>), resulting in the L31S substitution, is associated with dominant resistance to Cry1Ac in a major pest, <em>Helicoverpa armigera</em>. Previous studies using CRISPR/Cas9 technique have demonstrated that knockin of the <em>HaTSPAN1</em> T92C mutation confers a 125-fold resistance to Cry1Ac in the susceptible SCD strain of <em>H. armigera</em>. In this study, we employed the <em>piggyBac</em> transposon system to create two transgenic <em>H. armigera</em> strains based on SCD: one expressing the wild-type <em>HaTSPAN1</em> gene (SCD-TSPANwt) and another expressing the T92C mutant form of <em>HaTSPAN1</em> (SCD-TSPANmt). The SCD-TSPANmt strain exhibited an 82-fold resistance to Cry1Ac compared to the recipient SCD strain, while the SCD-TSPANwt strain remained susceptible. The Cry1Ac resistance followed an autosomal dominant inheritance mode and was genetically linked with the transgene locus in the SCD-TSPANmt strain of <em>H. armigera</em>. Our results further confirm the causal association between the T92C mutation of <em>HaTSPAN1</em> and dominant resistance to Cry1Ac in <em>H. armigera</em>. Additionally, they suggest that the <em>piggyBac</em>-mediated transformation system we used in <em>H. armigera</em> is promising for functional investigations of candidate Bt resistance genes from other lepidopteran pests.</p></div>","PeriodicalId":19828,"journal":{"name":"Pesticide Biochemistry and Physiology","volume":null,"pages":null},"PeriodicalIF":4.2000,"publicationDate":"2024-08-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"piggyBac-based transgenic Helicoverpa armigera expressing the T92C allele of the tetraspanin gene HaTSPAN1 confers dominant resistance to Bacillus thuringiensis toxin Cry1Ac\",\"authors\":\"Lin Li, Xinru Pang, Chenyang Wang, Yihua Yang, Yidong Wu\",\"doi\":\"10.1016/j.pestbp.2024.106096\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><p>Transgenic crops producing insecticidal proteins from <em>Bacillus thuringiensis</em> (Bt) have revolutionized pest control. However, the evolution of resistance by target pests poses a significant threat to the long-term success of Bt crops. Understanding the genetics and mechanisms underlying Bt resistance is crucial for developing resistance detection methods and management tactics. The T92C mutation in a tetraspanin gene (<em>HaTSPAN1</em>), resulting in the L31S substitution, is associated with dominant resistance to Cry1Ac in a major pest, <em>Helicoverpa armigera</em>. Previous studies using CRISPR/Cas9 technique have demonstrated that knockin of the <em>HaTSPAN1</em> T92C mutation confers a 125-fold resistance to Cry1Ac in the susceptible SCD strain of <em>H. armigera</em>. In this study, we employed the <em>piggyBac</em> transposon system to create two transgenic <em>H. armigera</em> strains based on SCD: one expressing the wild-type <em>HaTSPAN1</em> gene (SCD-TSPANwt) and another expressing the T92C mutant form of <em>HaTSPAN1</em> (SCD-TSPANmt). The SCD-TSPANmt strain exhibited an 82-fold resistance to Cry1Ac compared to the recipient SCD strain, while the SCD-TSPANwt strain remained susceptible. The Cry1Ac resistance followed an autosomal dominant inheritance mode and was genetically linked with the transgene locus in the SCD-TSPANmt strain of <em>H. armigera</em>. Our results further confirm the causal association between the T92C mutation of <em>HaTSPAN1</em> and dominant resistance to Cry1Ac in <em>H. armigera</em>. Additionally, they suggest that the <em>piggyBac</em>-mediated transformation system we used in <em>H. armigera</em> is promising for functional investigations of candidate Bt resistance genes from other lepidopteran pests.</p></div>\",\"PeriodicalId\":19828,\"journal\":{\"name\":\"Pesticide Biochemistry and Physiology\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":4.2000,\"publicationDate\":\"2024-08-20\",\"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/S0048357524003298\",\"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/S0048357524003298","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}
piggyBac-based transgenic Helicoverpa armigera expressing the T92C allele of the tetraspanin gene HaTSPAN1 confers dominant resistance to Bacillus thuringiensis toxin Cry1Ac
Transgenic crops producing insecticidal proteins from Bacillus thuringiensis (Bt) have revolutionized pest control. However, the evolution of resistance by target pests poses a significant threat to the long-term success of Bt crops. Understanding the genetics and mechanisms underlying Bt resistance is crucial for developing resistance detection methods and management tactics. The T92C mutation in a tetraspanin gene (HaTSPAN1), resulting in the L31S substitution, is associated with dominant resistance to Cry1Ac in a major pest, Helicoverpa armigera. Previous studies using CRISPR/Cas9 technique have demonstrated that knockin of the HaTSPAN1 T92C mutation confers a 125-fold resistance to Cry1Ac in the susceptible SCD strain of H. armigera. In this study, we employed the piggyBac transposon system to create two transgenic H. armigera strains based on SCD: one expressing the wild-type HaTSPAN1 gene (SCD-TSPANwt) and another expressing the T92C mutant form of HaTSPAN1 (SCD-TSPANmt). The SCD-TSPANmt strain exhibited an 82-fold resistance to Cry1Ac compared to the recipient SCD strain, while the SCD-TSPANwt strain remained susceptible. The Cry1Ac resistance followed an autosomal dominant inheritance mode and was genetically linked with the transgene locus in the SCD-TSPANmt strain of H. armigera. Our results further confirm the causal association between the T92C mutation of HaTSPAN1 and dominant resistance to Cry1Ac in H. armigera. Additionally, they suggest that the piggyBac-mediated transformation system we used in H. armigera is promising for functional investigations of candidate Bt resistance genes from other lepidopteran pests.
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.