Pesticide Biochemistry and Physiology最新文献

{"title":"Exploring mechanisms of resistance to fludioxonil in Colletotrichum fructicola","authors":"Hafiz Muhammad Usman ,&nbsp;Mohammad Mazharul Karim ,&nbsp;Ayesha Kanwal ,&nbsp;Qin Tan ,&nbsp;Muhammad Dilshad Hussain ,&nbsp;Wei-Xiao Yin ,&nbsp;Yong Wang ,&nbsp;Chao-Xi Luo","doi":"10.1016/j.pestbp.2024.106284","DOIUrl":"10.1016/j.pestbp.2024.106284","url":null,"abstract":"<div><div><em>Colletotrichum fructicola</em> is one of the most important species causing peach anthracnose around the world, including China. Fludioxonil has been effectively applied to control anthracnose disease as well as several important fungal diseases such as gray mold, leaf blight, early blight, corn stem rot, peanut root rot, rice evil seedling disease, and other diseases transmitted through seeds or soil. In this study, 39<em>C. fructicola</em> isolates were collected from different locations in Guizhou Province and Guangdong Province. A sensitive isolate of <em>C. fructicola</em>, previously thought to be naturally resistant to fludioxonil, was unexpectedly detected. No cross-resistance was found for fludioxonil with procymidone, prochloraz, and pyraclostrobin. Significant differences were observed between sensitive and resistant isolates in terms of mycelial growth rate and osmotic sensitivity experiments (4 %, 6 %, and 8 % NaCl), but no significant difference was found for sporulation. A novel mutation I880V was detected in the Os1 protein from one sensitive isolate. Molecular docking was used to explain the reversal of inherent resistance to sensitivity of <em>C. fructicola</em> to fludioxonil. Results showed that the wild type Os1 protein was docked against fludioxonil with a binding energy of −6.8 kj/mol, while it increased to −6.6 kj/mol between the mutated protein and fludioxonil. At the same time, different interactions were observed between wild type and mutated proteins with fludioxonil. These results suggest that the I880V mutation in the Os1 protein changed the conformation of the binding pocket, potentially leading to the reversal from resistance to sensitivity to fludioxonil. These findings are remarkable in demonstrating the fludioxonil resistance mechanism, and further studies such as genetic transformation and a range of molecular investigations are necessary to validate resistance mechanisms, elucidate the molecular pathways involved, and develop effective disease management strategies.</div></div>","PeriodicalId":19828,"journal":{"name":"Pesticide Biochemistry and Physiology","volume":"208 ","pages":"Article 106284"},"PeriodicalIF":4.2,"publicationDate":"2025-01-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143095556","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Identification of an α-galactosidase with high affinity and synergistic activity against Bacillus thuringiensis App6Aa2 toxin in Bursaphelenchus xylophilus","authors":"Yajie Guo ,&nbsp;Yanyue Zhou ,&nbsp;Jun Li ,&nbsp;Mingqing Weng ,&nbsp;Yunzhu Sun ,&nbsp;Songqing Wu ,&nbsp;Chunlan Lian","doi":"10.1016/j.pestbp.2024.106282","DOIUrl":"10.1016/j.pestbp.2024.106282","url":null,"abstract":"<div><div>The App6Aa2 toxin, derived from <em>Bacillus thuringiensis</em>, is nematicidal and highly toxic to <em>Bursaphelenchus xylophilus</em>. Receptors play a critical role in the mechanism of <em>B. thuringiensis</em> crystal toxin toxicity, yet the specific binding receptors for App6Aa2 in <em>B. xylophilus</em> have not been identified. This study identified a GPI-anchored protein, <em>α</em>-galactosidase (<em>Bx</em>Gal), in <em>B. xylophilus</em> as a potential binding protein. Western blotting and ELISA assays confirmed a high binding affinity between <em>Bx</em>Gal and App6Aa2 (<em>Kd</em> = 24.5 ± 11.2 nM). Remarkably, combining App6Aa2 with the <em>Bx</em>Gal protein produced a synergistic effect, significantly increasing nematode mortality from 25 % to 81 % (<em>P</em> &lt; 0.05), thereby enhancing the toxicity of App6Aa2 against <em>B. xylophilus</em>. Besides, RNAi silencing of <em>Bx</em>Gal in nematodes had no specific effect on App6Aa2 toxicity at high concentrations, while mortality increased slightly at lower concentrations. These findings indicate that <em>Bx</em>Gal is a high-affinity binding protein for App6Aa2; it does not function as the primary receptor and warrants further investigation into its role in modulating nematode susceptibility to App6Aa2.</div></div>","PeriodicalId":19828,"journal":{"name":"Pesticide Biochemistry and Physiology","volume":"208 ","pages":"Article 106282"},"PeriodicalIF":4.2,"publicationDate":"2024-12-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143155709","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Involvement of AhR/ARNT in tolerance to deltamethrin by regulating the expression of TcGSTs6 in Tribolium castaneum","authors":"Zhichao Luo ,&nbsp;Haoting Chen ,&nbsp;Daojie Guan ,&nbsp;Chen Hang ,&nbsp;Zhichao Wang ,&nbsp;Huichen Ge ,&nbsp;Kun Qian ,&nbsp;Jianjun Wang","doi":"10.1016/j.pestbp.2024.106281","DOIUrl":"10.1016/j.pestbp.2024.106281","url":null,"abstract":"<div><div>It is well known that the transcription factors aryl hydrocarbon receptor (AhR) and aryl hydrocarbon receptor nuclear translocator (ARNT) play important roles in the detoxification of xenobiotic compounds including insecticides. In this study, <em>TcAhR</em> and <em>TcARNT</em> were cloned from <em>Tribolium castaneum</em>. RT-qPCR analysis revealed that <em>TcAhR</em> and <em>TcARNT</em> were highly expressed in brain and epidermis of female adults and upregulated after exposure of the adults to deltamethrin. Knockdown of <em>TcAhR</em> and <em>TcARNT</em> increased the susceptibility of <em>T. castaneum</em> to deltamethrin and decreased the enzymatic activity of Glutathione S-transferase (GST). Consistently, knockdown of <em>TcAhR</em> and <em>TcARNT</em> led to the downregulation of <em>TcGSTs6</em> and suppressed the induction of <em>TcGSTs6</em> by deltamethrin treatment. Increased susceptibility of <em>T. castaneum</em> to deltamethrin was also observed after knockdown of <em>TcGSTs6</em>. Further dual-luciferase reporter assay revealed that co-overexpression of <em>TcAhR</em> and <em>TcARNT</em> in Sf9 cells significantly enhanced the promoter activity of <em>TcGSTs6</em>. These results provided insights into the function of insect AhR and ARNT as well as the regulatory mechanisms of insect GSTs.</div></div>","PeriodicalId":19828,"journal":{"name":"Pesticide Biochemistry and Physiology","volume":"208 ","pages":"Article 106281"},"PeriodicalIF":4.2,"publicationDate":"2024-12-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143095549","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Insecticidal activity of two Pelargonium essential oils and head transcriptome analysis of stored-product pest Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae) in response to citronellyl formate fumigation","authors":"Guang-Wen Fan ,&nbsp;Pei Wang ,&nbsp;Yang-Shan Liu ,&nbsp;Yu-Li Sang ,&nbsp;Nan Liu ,&nbsp;Yan-Jun Hao","doi":"10.1016/j.pestbp.2024.106278","DOIUrl":"10.1016/j.pestbp.2024.106278","url":null,"abstract":"<div><div><em>Tribolium castaneum</em> (Herbst) is one of the most common stored-product pests, causing enormous economic losses and developing widespread resistance to chemical insecticides. Natural products derived from essential oils (EOs) are well-known for insecticidal activity against agricultural pests, especially in the management of stored-product pests. In this study, the chemical constituents and repellent, contact and fumigation toxicity activities of two <em>Pelargonium</em> EOs against <em>T. castaneum</em> were evaluated. Moreover, the fumigation mechanism of citronellyl formate was assessed by head transcriptome sequencing and RNA interference (RNAi). A total of 28 and 39 compounds were identified by gas chromatography-mass spectrometry, accounting for 98.58 and 97.33 % of <em>P. roseum</em> and <em>P. asperum</em> EOs, and the major components were citronellol, (1<em>S</em>)-(1)-<em>β</em>-pinene and citronellyl formate. (1<em>S</em>)-(1)-<em>β</em>-Pinene exhibited strong contact toxicity activity (LD₅₀ = 19.72 <em>μ</em>g/adult), while citronellyl formate exhibited strong repellent and fumigation toxicity activities, with the LC₅₀ value of 11.93 mg/L air. Under the stress of citronellyl formate, 1222 differentially expressed genes were identified, including 554 up-regulated and 668 down-regulated genes, which was further verified by qRT-PCR. Among odorant-binding proteins (OBPs), only <em>TcGOBP70</em> was up-regulated, suggesting that <em>GOBP70</em> is more likely to be involved in the defense of <em>T. castaneum</em> by recognizing, binding and transporting citronellyl formate. Additionally, RNAi against <em>TcGOBP70</em> dramatically increased the contact and fumigation toxicity activities of citronellyl formate, with mortality rates of 73 and 75 %, respectively. Therefore, our findings not only provided theoretical bases for the comprehensive utilization of the <em>Pelargonium</em> species but also established potential targets for controlling <em>T. castaneum</em> adults.</div></div>","PeriodicalId":19828,"journal":{"name":"Pesticide Biochemistry and Physiology","volume":"208 ","pages":"Article 106278"},"PeriodicalIF":4.2,"publicationDate":"2024-12-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143095548","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"An in-depth analysis of the effects of excessive acetochlor exposure on chicken liver health","authors":"Yue Zhang,&nbsp;Lulu Hou,&nbsp;Tiantian Guo,&nbsp;Hongmin Lu,&nbsp;Xin Zhang,&nbsp;Mingwei Xing","doi":"10.1016/j.pestbp.2024.106280","DOIUrl":"10.1016/j.pestbp.2024.106280","url":null,"abstract":"<div><div>Acetochlor, a commonly used herbicide, poses significant risks to ecosystem and organism health through contamination of the food chain. Despite its widespread use, there is a lack of comprehensive studies on its toxicological effects on avian species. This study investigates the impact of environmental acetochlor exposure on chicken liver health using metabolomics analysis and histopathological techniques. Microscopic examination revealed autophagy-like structures and endoplasmic reticulum (ER) expansion, with significant effects observed at higher exposure levels. Biochemical analysis and metabolomics also demonstrated acetochlor-induced ferroptosis, highlighting disruptions in liver function. Further, in vitro studies revealed that acetochlor stimulates autophagy, which regulates ferroptosis via ferritin degradation, mediated through the ER-CaMKII pathway. These findings emphasize the importance of understanding the molecular mechanisms involved in acetochlor toxicity, particularly the role of the Ca²⁺/CaMKII pathway, ER stress, and autophagy in ferroptosis. The study contributes to a deeper understanding of how environmental contaminants affect avian species, providing critical insights for better herbicide risk assessment, pollution control, and sustainable agricultural practices.</div></div>","PeriodicalId":19828,"journal":{"name":"Pesticide Biochemistry and Physiology","volume":"208 ","pages":"Article 106280"},"PeriodicalIF":4.2,"publicationDate":"2024-12-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143155706","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Peptide neurotoxins affecting insect voltage-gated calcium channels and possessing insecticidal toxicity: Two ω-Atypitoxins from Calommata signata","authors":"Yang-yang Yan,&nbsp;Kan Wang,&nbsp;Jing-ting Wang,&nbsp;Qian-qian Han,&nbsp;Zhen Zhang,&nbsp;Na Yu,&nbsp;Ze-Wen Liu","doi":"10.1016/j.pestbp.2024.106279","DOIUrl":"10.1016/j.pestbp.2024.106279","url":null,"abstract":"<div><div>Spider peptide toxins, as potent insecticides, distinguish them from current insecticides, and thus expand the range of viable molecular targets of insecticides in insect pests. This study reports the isolation of two neurotoxic peptides, ω-Atyptoxin-Cs2a (Cs2a) and ω-Atyptoxin-Cs2b (Cs2b), from the venom gland of the burrowing spider <em>Calommata signata</em>. These peptides exhibit strong insecticidal activity against two insect pests with agricultural importance, <em>Nilaparvata lugens</em> and <em>Spodoptera frugiperda</em>. Both toxins demonstrated rapid and significant lethality, with LD₅₀ values of 1.083 ± 0.120 nmol/g and 0.949 ± 0.079 nmol/g for <em>N. lugens</em> and 1.035 ± 0.114 nmol/g and 0.998 ± 0.081 nmol/g for <em>S. frugiperda</em> within 24 h. Using the whole-cell patch-clamp electrophysiology, Cs2a and Cs2b were identified as inhibitors of high-voltage-activated calcium channels in cockroach DUM neurons, with IC₅₀ values of 0.504 ± 0.078 μM and 0.411 ± 0.053 μM, respectively. These findings show that Cs2a and Cs2b are effective calcium channel blockers with potential to develop bioinsecticides, offering selective toxicity toward insect pests.</div></div>","PeriodicalId":19828,"journal":{"name":"Pesticide Biochemistry and Physiology","volume":"208 ","pages":"Article 106279"},"PeriodicalIF":4.2,"publicationDate":"2024-12-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143099736","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of a novel anti-microsporidia strategy by inhibiting parasite and host glucose metabolism","authors":"Zhanqi Dong ,&nbsp;Qin Wu ,&nbsp;Pengcheng Zhang ,&nbsp;Wenxuan Fang ,&nbsp;Xiaocui Lei ,&nbsp;Boyuan Deng ,&nbsp;Nan Hu ,&nbsp;Peng Chen ,&nbsp;Xuhua Huang ,&nbsp;Cheng Lu ,&nbsp;Minhui Pan","doi":"10.1016/j.pestbp.2024.106276","DOIUrl":"10.1016/j.pestbp.2024.106276","url":null,"abstract":"<div><div>Microsporidia are obligate intracellular parasites that infect most types of animals. Exploring how microsporidia utilize energy substrates in infected host cells is important for human health and the development of the agricultural economy. In this study, transcriptomics was used to systematically analyze the enriched pathways involving ATP/ADP transporters and energy metabolism during the schizont proliferation period of <em>Nosema bombycis</em>. A <em>Nosema bombycis</em> ADP/ATP carrier 1 (NbAAC1) protein function characteristics of the adenine nucleotide translocase family were identified after infection with <em>N. bombycis</em>. NbAAC1 could inhibit ATP production and affect <em>Nosema bombycis</em> proliferation based on RNA interference <em>in vivo</em> and <em>in vitro</em>. Meanwhile, an effective gene-edited line targeted editing of the <em>Bombyx mori</em> hexokinase (<em>BmHXK</em>) gene of the host glycolytic metabolism pathway could inhibit <em>N. bombycis</em> infection was established. These findings provide new therapeutic approaches to controlling microsporidia infections by inhibiting intracellular parasitic fungi and host energy metabolism.</div></div>","PeriodicalId":19828,"journal":{"name":"Pesticide Biochemistry and Physiology","volume":"208 ","pages":"Article 106276"},"PeriodicalIF":4.2,"publicationDate":"2024-12-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143155707","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"N-acetylglucosamine sensor, Ngs1 contributes to Beauveria bassiana vegetative growth, oxidative phosphorylation, fungal development, and cell wall integrity during entomopathogen-insect interaction","authors":"Zhou Cui ,&nbsp;Wu-Wei-Jie Yang ,&nbsp;Zhi-Hao Yang ,&nbsp;Long-Bin Zhang ,&nbsp;Yi Guan","doi":"10.1016/j.pestbp.2024.106273","DOIUrl":"10.1016/j.pestbp.2024.106273","url":null,"abstract":"<div><div><em>N</em>-acetylglucosamine (GlcNAc), a key component of fungal cell walls and insect cuticles, is an important signal to activate fungal response during entomopathogen-insect interactions. Research on Ngs1, the only identified GlcNAc sensor and transducer, has been primarily restricted to <em>Candida</em> species. Although our previous work identified an Ngs1 homology in <em>Beauveria bassiana</em>, its physiological functions in entomopathogenic fungi remain largely unexplored. In this study, we unveiled the sub-localization of Ngs1 in the nucleolus. Further transcriptomic analysis revealed that Ngs1 plays a crucial role in vegetative growth, fungal development, and cell-wall construction by acting as a transcriptional mediator, particularly influencing carbon metabolism in response to insect cuticle stimulation. The absence of <em>Ngs1</em> compromised vegetative growth across various carbon sources by downregulating expressions of key catalytic enzymes. Conversely, <em>Ngs1</em> deficiency enhanced transcription levels of oxidative phosphorylation, leading to increased ATP and reactive oxygen species (ROS) production. Despite higher ATP levels, <em>Ngs1</em>-deletion mutants exhibited reduced asexual development and hyphal germination, primarily due to the function of <em>Ngs1</em> in the central developmental pathway and Brg1/Nrg1-dependent pathway. Additionally, the downregulation of N-glycan biosynthesis in Δ<em>Ngs1</em> diminished cell wall components, resulting in decreased cell wall resistance to lysis and impaired fungal development. These findings advance our understanding of the regulatory role of Ngs1 in <em>B. bassiana</em> during host interactions and provide a theoretical foundation for engineering fungi to maintain or even enhance pesticidal activity.</div></div>","PeriodicalId":19828,"journal":{"name":"Pesticide Biochemistry and Physiology","volume":"208 ","pages":"Article 106273"},"PeriodicalIF":4.2,"publicationDate":"2024-12-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143099745","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A UDP-glucuronosyltransferase gene UGT379A1 involved in detoxification of lufenuron in Diaphorina citri","authors":"Ze-Hao Huang ,&nbsp;Qi Pan ,&nbsp;Zhen Wu ,&nbsp;Yu-Chao Shen ,&nbsp;Si-Chen Li ,&nbsp;Qi-Qi Yang ,&nbsp;Shao-Hui Zhang ,&nbsp;Shuang Lei ,&nbsp;Yang-Yang Cui ,&nbsp;Li-Li Ding ,&nbsp;Shi-Jiang Yu ,&nbsp;Liu Liu ,&nbsp;Lin Cong ,&nbsp;Bing-Hai Lou ,&nbsp;Xue-Feng Wang ,&nbsp;Chun Ran ,&nbsp;Ling Liao","doi":"10.1016/j.pestbp.2024.106260","DOIUrl":"10.1016/j.pestbp.2024.106260","url":null,"abstract":"<div><div>The Asian citrus psyllid, <em>Diaphorina citri</em> Kuwayama (Hemiptera: Psyllidae), is a devastating bacterial disease of commercial citrus. Presently, uridine diphosphate (UDP)-glycosyltransferases (UGTs), have been linked to the detoxification of pesticides, were known as phase II enzymes in the detoxification process. However, the role of UGTs in detoxification of lufenuron in <em>Diaphorina citri</em> is unknown. In this study, we identified a UGT gene, <em>UGT379A1</em>, which was significantly up-regulated under the exposure of lufenuron. The knockdown of <em>UGT379A1</em> increased the susceptibility of <em>D. citri</em> to lufenuron. <em>In vitro</em> metabolism and Dixon plot analysis indicated that <em>UGT379A1</em> could deplete lufenuron by sequestration. These results showed that <em>UGT379A1</em> was involved in the lufenuron detoxification, which provides a theoretical basis for the prevention and control of <em>D. citri</em>.</div></div>","PeriodicalId":19828,"journal":{"name":"Pesticide Biochemistry and Physiology","volume":"208 ","pages":"Article 106260"},"PeriodicalIF":4.2,"publicationDate":"2024-12-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143099735","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A streamlined cloning and toxicity evaluation strategy of toxin-antitoxin systems in insect cells","authors":"Yunfei Li,&nbsp;Shizhe Hu,&nbsp;Jing Wang,&nbsp;Liping Lv,&nbsp;Xueting Zhao,&nbsp;Hanbing Li,&nbsp;Weihua Ma,&nbsp;Hongxia Hua,&nbsp;Zhihui Zhu","doi":"10.1016/j.pestbp.2024.106261","DOIUrl":"10.1016/j.pestbp.2024.106261","url":null,"abstract":"<div><div>Toxin-antitoxin (TA) systems, which are prevalent in bacteria and archaea, have significant potential applications in eukaryotic cells due to their characteristic ability to inhibit cell growth. In particular, type II TA systems, where both the toxin and antitoxin are proteins, can serve as lethal and rescue genes in genetic pest management. However, traditional methods of cloning these systems often encounter difficulties due to cryptic promoter driven expression of toxins during the cloning process. To address this issue, we have developed a novel cloning method that incorporates an additional prokaryotic promoter to express the antitoxin, thus preventing the deleterious effects of toxin expression in bacterial cells. This method allows for efficient and straightforward construction of toxin vectors and the rapid screening of effective TA systems in insect cells. Our study focused on evaluating the toxicity of various type II TA systems in Sf9 insect cells. We cloned and tested toxins and antitoxins from <em>Escherichia coli</em>, <em>Streptococcus pneumoniae</em>, and <em>Rickettsia rickettsii</em>. The toxicity of each system was assessed by the number of red fluorescent cells post-transfection. Our results demonstrated that toxins such as MazF (<em>E. coli</em>-2782), RelE (<em>Spn</em>-1223), and RelE (<em>Spn</em>-1104) exhibited high levels of toxicity in Sf9 cells, while other toxins showed varying degrees of effectiveness. The corresponding antitoxins were able to neutralize the toxins in a dose-dependent manner, although some antitoxins, like MnT (<em>R. felis</em>-0357) and MazE (<em>E. coli</em>-4224), were ineffective in insect cells. This study highlights the potential for applying type II TA systems in genetic pest management and other eukaryotic applications. Our novel cloning approach provides a robust platform for the rapid and efficient evaluation of TA systems, paving the way for future research and application in pest control and beyond.</div></div>","PeriodicalId":19828,"journal":{"name":"Pesticide Biochemistry and Physiology","volume":"208 ","pages":"Article 106261"},"PeriodicalIF":4.2,"publicationDate":"2024-12-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143156003","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}