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The dynamic changes of genes revealed that persistently overexpressed genes drive the evolution of cyflumetofen resistance in Tetranychus cinnabarinus

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Insect Science 2023年第4期30卷 1129-1148页

作者： Kaiyang Feng Jialu Liu Mingyu Zhao Zhixin Jiang Peilin Liu Peng Wei Wei Dou Lin He Key Laboratory of Entomology and Pest Control Engineering College of Plant ProtectionSouthwest UniversityChongqingChina Academy of Agricultural Sciences Southwest UniversityChongqingChina State Cultivation Base of Crop Stress Biology for Southern Mountainous Land of Southwest University Southwest UniversityChongqingChina key Scientific Research Base of pest and Mold control of Heritage Collection(Chongqing China Three Gorges Museum) State Administration of Cultural HeritageChongqingChina

Changes in gene expression are associated with the evolution of pesticide resis-tance in arthropods.In this study,transcriptome sequencing was performed in 3 different resistance levels(low,L;medium,M;and high,H)of cyflumetofen-resistant strain(YN-CyR).A total of 1685 genes,including 97 detoxification enzyme genes,were upregulated in all 3 stages,of which 192 genes,including 11 detoxification enzyme genes,showed a continuous increase in expression level with resistance development(L to H).RNA in-terference experiments showed that overexpression of 7 genes(CYP392A1,TcGSTd05,CCE06,CYP389A1,TcGSTz01,CCE59,and CYP389C2)is involved in the development of cyflumetofen resistance in Tetranychus cinnabarinus.The recombinant CYP392A1 can effectively metabolize cyflumetofen,while CCE06 can bind and sequester cyflumetofen in vitro.We compared 2 methods for rapid screening of resistance molecular markers,in-cluding short-term induction and 1-time high-dose selection.Two detoxification enzyme genes were upregulated in the field susceptible strain(YN-S)by induction with 20%lethal concentration(LC2o)of cyflumetofen.However,16 detoxification enzyme genes were up-regulated by 1-time selection with LCso of cyflumetofen.Interestingly,the 16 genes were overexpressed in all 3 resistance stages.These results indicated that 1685 genes that were upregulated at the L stage constituted the basis of cyflumetofen resistance,of which 192 genes in which upregulation continued to increase were the main driving force for the de-velopment of resistance.Moreover,the 1-time high-dose selection is an efficient way to rapidly obtain the resistance-related genes that can aid in the development of resistance markers and resistance management in mites.

关键词： change rules cyflumetofen detoxification enzyme genes resistance molec-ular markers Tetranychus cinnabarinus

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Transcription factors CncC and Maf connect the molecular network between pesticide resistance and resurgence of pest mites

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Insect Science 2022年第3期29卷 801-816页

作者： Guang-Mao Shen Shi-Yuan Ou Chuan-Zhen Li Kai-Yang Feng Jin-Zhi Niu Michael J.Adang Lin He Key Laboratory of Entomology and Pest Control Engineering College of Plant ProtectionSouthwest UniversityChongqingChina Academy of Agricultural Sciences Southwest UniversityChongqingChina State Cultivation Base of Crop Stress Biology for Southern Mountainous Land of Southwest University Southwest UniversityChongqingChina Department of entomology University of GeorgiaAthensGeorgiaUSA

pesticide resistance and resurgence are serious problems often occurring simultaneously in the field.In our long-term study of a fenpropathrin-resistant strain of Tetranychus cinnabaribus,enhancement of detoxification and modified fecundity mechanisms were both observed.Here we investigate the network across these two mechanisms and find a key node between resistance and resurgence.We show that the ecdysone pathway is involved in regulating the fecundity of T.cinnabaribus.The concentration change of ecdysone is consistent with the fecundity curve;the concentration of ecdysone is higher in the fenpropathrin-resistant strain which has stronger fecundity.The enhancement of ecdysone is due to overexpression of two P450 genes(CYP314A1 and CYP315A1)in the ecdysone synthesis pathway.Silencing expression of these CYP genes resulted in lower concentration of ecdysone,reduced expression of vitellogenin,and reduced fecundity of T.cinnabaribus.The expression of CYP315A1 is regulated by transcription factors Cap-n-collar isoform C(CncC)and Musculoaponeurotic fibrosarcoma protein(Maf),which are involved in regulating other P450 genes functioning in detoxification of fenpropathrin in T.cinnabaribus.A similar regulation is established in citrus pest mite Panonychus citri showing that the CncC pathway regulates expression of PcCYP315Al,which affects mite fecundity.Transcription factors are activated to upregulate detoxification genes facilitating pesticide resistance,while the“one to multiple”regulation mode of transcription factors simultaneously increases expression of metabolic enzyme genes in hormone pathways and alters the physiology of pests.This is an important response of arthropods to pesticides which leads to resistance and population resurgence.

关键词： resistance resurgence Tetranychus cinnabarinus transcription factor

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Influence of various stressors on the expression of core genes of the small interfering RNA pathway in the oriental fruit fly, Bactrocera dorsalis

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Insect Science 2017年第3期24卷 418-430页

作者： Yi-Fei Xie Jin-Zhi Niu Xuan-Zhao Jiang Wen-Jia Yang Guang-Mao Shen Dong Wei Guy Smagghe Jin-Jun Wang Key Laboratory of Entomology and Pest Control Engineering College of Plant Protection Southwest University Chongqing China Department of Crop Protection Faculty of Bioscience Engineering Ghent University Ghent Belgium Hunan Academy of Forestry Changsha China

RNA interference （RNAi）-based technology has emerged as a potential tool for controlling insect pests, however, previous studies found that the efficiency of RNAi in Bactrocera dorsalis was variable. In nature, insects often meet various challenges, such as pathogen infections, extreme temperatures, lack of nutrition and heavy metals. To better understand the association of the stressors with efficiency of RNAi, in the current study we tested the expression of three core genes, dicer2 （Bddcr2）, r2d2 （Bdr2d2） and argonaute2 （Bdago2）, of the small interfering RNA （siRNA） pathway of B. dorsalis upon various stressors. Our results showed that all three genes were upregulated by the infection of invertebrate iridescent virus 6, which suggested a function of the siRNA pathway against viral infection. The loading of FeC13 could also increase the expression of Bddcr2. The treatments ofEscherichia coli, extremely high （40~C） and low （0~C） temperatures, as well as starvation, could negatively influence the expression of Bddcr2 and/or Bdago2. In total, our results showed that various stressors could influence the expression of core components ofB. dorsalis siRNA pathway. This highlights further speculation on the RNAi efficiency upon these stressors. Considering the complexity and variation of RNAi efficiency in different conditions, these results provide initial aspects in possible environmental stressors to influence the activity of the siRNA pathway, but the real impact of RNAi efficiency posed by these stressors requires further studies.

关键词： argonaute Bactrocera dorsalis dicer RNA interference R2D2 stressor

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Characterization of the Purified Glutathione S-transferases from Two Psocids Liposcelis bostrychophila and L.entomophila

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Agricultural Sciences in China 2010年第7期9卷 1008-1016页

作者： DOU Wei XIAO Li-sha NIU Jin-zhi JIANG Hong-bo WANG Jin-jun Key Laboratory of Entomology and Pest Control Engineering College of Plant Protection Southwest University Chongqing 400716 P.R. China

Glutathione S-transferases（GSTs） from Liposcelis bostrychophila Badonnel and L.entomophila（Enderlein）（Psocoptera：Liposcelididae） were purified by glutathione-agarose affinity chromatography,and characterized subsequently by their Michaelis-Menten kinetics toward the artificial substrates 1-chloro-2,4-dinitrobenzene（CDNB） and reduced glutathione（GSH）,respectively.The specific activity of the purified GST toward CDNB was 2.3-fold higher in L.bostrychophila than in L.entomophila.Though the specific activities of purified enzymes varied between the two species,the purification yields were similar.SDS-PAGE revealed one band at 23 kDa for both the species.GSTs of L.entomophila exhibited higher Michaelis-Menten constants（Km） but lower maximal velocity（Vmax） values than those of L.bostrychophila.The optimum pH for CDNB conjugation of L.bostrychophila and L.entomophila GSTs was 7.0 and 7.5,and optimum temperature was 35 and 40°C,respectively.Inhibition kinetics showed that cibacron blue,curcumin,bromosulfalein,ethacrynic acid,and carbosulfan had excellent inhibitory effects on GSTs in both species,but the inhibitory effects of beta-cypermethrin,fenpropathrin,tetraethylthiuram disulfide,and diethyl maleate were not significant.

关键词： GSTs purification psocids xenobiotic compounds

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Characterization of Domeless receptors and the role of Bd Domeless3 in anti-symbiont-like virus defense in Bactrocera dorsalis

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Journal of Integrative Agriculture 2024年第4期23卷 1274-1284页

作者： Wei Zhang Shaoyang Li Rong Li Jinzhi Niu Jinjun Wang Key Laboratory of Entomology and Pest Control Engineering College of Plant ProtectionSouthwest UniversityChongqing 400715China key laboratory of Agricultural Biosafety and Green Production of Upper Yangtze River Ministry of Education/International Joint Laboratory of China-Belgium on Sustainable Crop Pest ControlAcademy of Agricultural SciencesSouthwest UniversityChongqing 400715China

The Janus kinase/signal transducers and activators of transcription(JAK/STAT)signaling pathway play a pivotal role in innate immunity.Among invertebrates,Domeless receptors serve as the key upstream regulators of this pathway.In our study on Bactrocera dorsalis,we identified three cytokine receptors:BdDomeless1,BdDomeless2,and BdDomeless3.Each receptor encompasses five fibronectin-type-III-like(FN III)extracellular domains and a transmembrane domain.Furthermore,these receptors exhibit the increased responsiveness to diverse pathogenic challenges.Notably,only BdDomeless3 is upregulated during symbiont-like viral infections.Moreover,silencing BdDomeless3 enhanced the infectivity of Bactrocera dorsalis cripavirus(BdCV)and B.dorsalis picorna-like virus(BdPLV),underscoring BdDomeless3’s crucial role in antiviral defense of B.dorsalis.Following the suppression of Domeless3 expression,six antimicrobial peptide genes displayed decreased expression,potentially correlating with the rise in viral infectivity.To our knowledge,this is the first study identifying cytokine receptors associated with the JAK/STAT pathway in tephritid flies,shedding light on the immune mechanisms of B.dorsalis.

关键词： Bactrocera dorsalis JAK/STAT pathway Domeless receptors antiviral immunity symbiont-like virus

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CRISPR/Cas9-mediated knockout of period reveals its function in the circadian rhythms of the diamondback moth Plutella xylostella

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Insect Science 2023年第3期30卷 637-649页

作者： Danfeng Wang Jing Chen Yao Yuan Lingqi Yu Guang Yang Wenfeng Chen State key laboratory of Ecological pest control for Fujian and Taiwan Crops Institute of Applied EcologyFujan Agriculture and Forestry UniversityFuzhouChina Joint International Research laboratory of Ecological pest control Ministry of EducationFuzhouChina key laboratory of Integrated pest Management for Fujian-Taiwan Crops Ministry of AgricultureFuzhouChina key laboratory of Green control of Insect pests(Fujian Agriculture and Forestry University) Fujian Province UniversityFuzhouChina Ministerial and Provincial Joint Innovation Centre for Safety Production of Cross-Strait Crops Fujian Agriculture and Forestry UniversityFuzhouChina Institute of Life Sciences College of Biological Science and EngineeringFuzhou UniversityFuzhouChina

Circadian clocks control the rhythmicity of many behaviors and physiological features of insects.To study the circadian clock of the moth Plutella xylostella,we employed CRISPR/Cas9-mediated genome editing to investigate the effect of loss of the clock gene period on the circadian rhythms.P.xylostella harbors a single copy of period.Phylogenetic analysis showed that P.xylostella PERIOD is more homologous to mouse PERIOD than the PERIOD proteins from bees,flies,mosquitos,and many other Lepidoptera,such as Danaus plexippus and Bombyx mori.The circadian rhythms in adult locomotor activity were altered in the period knockout strain of P.xylostella under light–dark(LD)and continuous dark(DD)conditions.Under the LD cycle,the wild-type moths displayed nocturnal activity with activity peaking very early after lights off and quickly declining after lights on.In contrast,the period knockout strain had no peak in activity when the lights were turned off and exhibited steady activity throughout the hours of darkness.Interestingly,under DD conditions,our results showed that the locomotor rhythm can be maintained without period gene,but at a lower rhythmicity ratio than wild-type.In addition,knockout of period in P.xylostella changed circadian rhythms patterns related to pupal eclosion,mating,egg-laying,and egg hatching.Mechanistically,loss of PERIOD disrupted the molecular rhythm of period and changed the clock transcription rhythm in the heads of the moths under LD and DD conditions.Together,our study indicates that the PERIOD is required for normal expression of many behavioral rhythms in P.xylostella.

关键词： CRISPR/Cas9 circadian rhythm period Plutella xylostella

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Identification and characterization of odorant binding proteins in the diamondback moth,Plutella xylostella

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Insect Science 2021年第4期28卷 987-1004页

作者： Li-Jun Cai Li-Shuang Zheng Yu-Ping Huang Wei Xu Min-Sheng You State key laboratory of Ecological pest control for Fujian and Taiwan Crops Institute of Applied EcologyFujian Agriculture and Forestry UniversityFuzhouChina Joint International Research laboratory of Ecological pest control Ministry of EducationFuzhouChina key laboratory of Integrated pest Management for Fujian-Taiwan Crops Ministry of AgricultureFuzhouChina Agricultural Sciences College of ScienceHealthEngineering and EducationMurdoch UniversityMurdochAustralia

Odorant binding proteins(OBPs)are a group of soluble proteins functioning as odorant carriers in insect antennae,mouth parts and other chemosensory organs.However,multiple insect OBPs have been detected in other tissues and various functions have been proposed.Therefore,a detailed expression profile including stages,tissues and sexes where OBPs are expressed will assist in building the links to their potential functions,enhancing the functional studies of insect OBPs.Here,we identified 39 putative OBP genes from its genome and transcriptome sequences of diamondback moth(DBM),Plutella xylostella.The expression patterns of identified PxylOBPs were further investigated from eggs,larvae,pupae,virgin adults,mated adults,larval midgut,larval heads,adult antennae,adult heads and adult tarsi.Moreover,P.xylostella larvae and adults with and without host plants for 5 h were utilized to study the interactions between OBP expression and host plants.The results showed that most PxylOBPs were highly expressed in male and female adult antennae.The expression levels of certain PxyOBPs could be regulated by mating activities and feeding host plants.This study advances our knowledge of P.xylostella OBPs,which may help develop new strategies for more environmentally sustainable management of P.xylostella.

关键词： gene expression insect olfaction OBP qRT-PCR

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Identification,Characterization, and Expression of P450 Gene Encoding CYP6BQ13v2 from the Red Flour Beetle,Tribolium castaneum (Herbst) (Coleoptera:Tenebrionidae)

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Agricultural Sciences in China 2009年第10期8卷 1210-1218页

作者： XU Yong-qiang WANG Jin-jun JIANG Hong-bo DOU Wei TANG Pei-an AN Feng-ming Key Laboratory of Entomology and Pest Control Engineering College of Plant Protection Southwest University Chongqing 400716P.R. China

An allele of CYP6BQI3, named CYP6BQ 13v2 （GenBank accession no. FJ209361）, was isolated from the red flour beetle, Tribolium castaneum （Herbst）（Coleoptera： Tenebrionidae） by RT-PCR. The cDNA sequence of CYP6BQ13v2, 1 563 bp in length, contains an open reading frame of 1 554 nucleotides encoding a putative protein of 518 amino acid residues with a predicted molecular weight of 59.92 kDa and a theoretical pl of 7.60. The putative protein contains the classic hemebinding sequence motif F××G×××C×G （residues 456-465） conserved among all P450 enzymes as well as other characteristic motifs of all cytochrome P450s. It shares 98% identity with the previously published sequence of CYP6BQ13 （GenBank accession no. XP967146） from the T. castaneum genome project. Phylogenetic analysis of amino acid sequences from members of various P450 families indicated that there was closer phylogenetic relationship of CYP6BQ 13v2 with CYP302A1 and CYP307A1 mediating synthesis of the insect molting hormone, distant relationship with CYP6B1 metabolizing plant allelochemicals, CYP6D 1 linking to pyrethroid resistance and other members of CYP6 family. Expression test of the gene in the adults and immature stages of T. castaneum by quantitative real-time PCR revealed that CYP6BQ13v2 is expressed in all life stages investigated. The mRNA expression level in 1st instar larvae was 14.9- and 3.86-fold higher than those in pupae and adults, respectively. The CYP6BQ13v2 expression levels appeared in the order of 1st instar larvae, followed by 4th instar larvae, 7th instar larvae, adult, and pupae from high to low. The more bioinformation of CYP6BQ 13v2 was also analyzed.

关键词： Tribolium castaneum cytochrome P450 real-time PCR analysis bioinformatics

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Small GTPase Rab40C is upregulated by 20-hydroxyecdysone and insulin pathways to regulate ovarian development and fecundity

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Insect Science 2022年第6期29卷 1583-1600页

作者： Weiwei Zheng Fangyu Wu Yinhao Ye Tianran Li Zhenyu Zhang Hongyu Zhang key laboratory of Horticultural Plant Biology(MOE) Hubei Hongshan LaboratoryHubei Key Laboratory of Insect Resource Application and Sustainable Pest ControlInstitute of Urban and Horticultural EntomologyCollege of Plant Science and TechnologyHuazhong Agricultural UniversityWuhanChina

The insulin and 20-hydroxyecdysone(20E)pathways coordinately regulate in-sect vitellogenesis and ovarian development.However,the detailed molecular mechanisms such as the genes mediating the cooperation of the interaction of these 2 pathways in reg-ulating insect reproductive development are not well understood.In the present study,a small GTPase,Rab4oC,was identified from the notorious agricultural pest Bactrocera dorsalis.In addition to the well-known RAB domain,it also has a unique SOCS-box do-main,which is different from other Rab-GTPases.Moreover,we found that Rab40C was enriched in the ovaries of sexually mature females.RNA interference(RNAi)-mediated knockdown of BdRab40C resulted in a decrease in vitellogenin synthesis,underdevel-oped ovaries,and low fertility.Furthermore,depletion of insulin receptor InR or the het-erodimer receptor of 20E(EcRor USP)by RNAi significantlydecreased the transcription of BdRab40C and resulted in lower fecundity.Further studies revealed that the transcrip-tion of BdRab40C could be upregulated by the injection of insulin or 20E.These results indicate that Rab40C participates in the insulin and 20E pathways to coordinately regulate reproduction in B.dorsalis.Our results not only provide new insights into the insulin-and 20E-stimulated regulatory pathways controlling female reproduction in insects but also contribute to the development of potential eco-friendly strategies for pest control.

关键词： ecdysteriod fecundity insulin oriental fruit fly Rab40 vitellogenin

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World pest Day: Not for pests, But for People

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China CDC weekly 2020年第24期2卷 429-430页

作者： Jianguo Xu Chinese Academy of engineering Chinese pest control Association State key laboratory of Infectious Disease Prevention and control National Institute for Communicable Disease Control and PreventionChina CDC

World pest Day was inaugurated by the Chinese pest control Association and several organizations such as the Federation of Asian and Oceania pest Managers Associations,the National pest Management Association of US,Confederation of European pest Management Associations,World Health Organization(WHO)Collaborating Centre for Vector Surveillance and Management,and State key laboratory of Infectious Disease Prevention and control(China CDC).It was endorsed by 42 academicians from the Chinese Academy of Science and Chinese Academy of engineering,as well as 21 Members of the National Political Consultative Committee of China.June 6 is reserved annually to raise awareness among the public,government,and media about the important role that the pest control industry plays in protecting health,food,property and the environment from pest threats.

关键词： annually China. pest

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