Transcriptome analysis of Diaphorina citri Kuwayama in response to beta-cypermethrin stress
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摘要: 【目的】筛选高效氯氰菊酯胁迫下的柑橘木虱抗药性相关基因及其网络，探讨柑橘木虱应对拟除虫菊酯类杀虫剂的抗性机制，为柑橘木虱的抗药性治理提供科学依据。【方法】采用Illumina NovaSeq 6000测序系统对高效氯氰菊酯和空白对照处理的柑橘木虱成虫进行转录组测序，采用生物信息学软件Hisat2、DESeq2和clusterProfiler等对差异表达基因（Differentially expressed genes，DEGs）进行功能注释、分类以及参与的信号通路分析。【结果】从所检测的柑橘木虱转录组数据中筛选到显著DEGs 4596个，其中显著上调表达基因2900个、显著下调表达基因1696个。DEGs中有624个富集到KEGG的103条代谢通路，富集的通路主要涉及Hippo信号通路、核糖体、吞噬体、药物代谢、嘌呤代谢、硫胺素代谢和细胞凋亡等与昆虫抗药性紧密相关的7条代谢通路。与昆虫抗药性相关的DEGs 38个，涉及细胞色素P450、谷胱甘肽转移酶、钠离子通道、三磷酸腺苷合酶、细胞色素b、酸性磷酸酯酶、钙粘蛋白、糖基转移酶、脱羧酶和ABC转运蛋白等多个代谢抗性和靶标抗性基因。【结论】挖掘获得大量的柑橘木虱响应拟除虫菊酯类杀虫剂的抗药性相关基因，显著富集的通路主要涉及Hippo信号通路、核糖体、吞噬体和硫胺素代谢等与昆虫抗药性紧密相关的代谢通路，为研究柑橘木虱的抗药机制以及进行抗药性治理提供了丰富的数据基础。Abstract: 【Objective】 To screen Diaphorina citri Kuwayama resistance genes and their network regulation under the stress of beta-cypermethrin, and explore the resistance mechanism of D. citri to pyrethroid insecticides.【Method】 The Illumina NovaSeq 6000 sequencing system was used to sequence the transcriptome of the adult citrus psyllids treated with beta-cypermethrin and blank control. The differentially expressed genes(DEGs)were functionally annotated and classified using bioinformatics softwares such as HISAT2,DESeq2 and clusterProfiler,as well as the signal pathways analysis involved.【Result】 A total of 4596 significant DEGs genes were screened in the citrus psyllid transcriptome data,in which 2900 were significantly up-regulated and 1696 were significantly down-regulated. Among the DEGs,624 were enriched in 103 different metabolic pathways in KEGG. Seven enriched pathways of insect resistance mainly involved Hippo signaling pathway,ribosome,phagosome,drug metabolism,purine metabolism,thiamine metabolism,apoptosis,etc. Thirty-eight DEGs were related to insect drug resistance,involving multiple metabolic resistance and target resistance genes,including cytochrome oxidase,glutathione transferase,sodium ion channel,adenosine triphosphate synthase,cytochrome b,acid phosphoesterase,cadherin,glycosyltransferase,decarboxylase,ABC transporter,etc.【Conclusion】In this study,a lot of drug resistance-related genes of D. citri in response to pyrethroid insecticides are mined,and four significantly enriched metabolic pathways are closely related to insect resistance,such as Hippo signaling pathway,ribosome,phagosome,and thiamine metabolism,which provides a rich data basis for studying the drug resistance mechanism of citrus psyllid and for drug resistance management.
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Keywords:
- Diaphorina citri Kuwayama /
- beta-cypermethrin /
- drug resistance /
- transcriptome
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陈诚, 龚椿营, 王日远, 宋春暖, 徐汉福. 2016. 昆虫Hippo信号通路研究概述[J]. 蚕业科学, 42(1):152-161.[Chen C,Gong C Y, Wang R Y, Song C N, Xu H F. 2016. Research summary on the Hippo signaling pathway in insects[J].Science of Sericulture, 42(1):152-161.]doi: 10.13441/j.cnki.cykx.2016.01.022. 陈皓, 彭威, 韩宝瑜. 2020. 功能基因组技术在昆虫抗药性研究中的应用展望[J]. 植物保护学报, 47(2):223-233.[Chen H, Peng W, Han B Y. 2020. Application prospects of functional genome analysis technologies in insecticide resistance researches[J]. Journal of Plant Protection, 47(2):223-233.]doi: 10.13802/j.cnki.zwbhxb.2020.2019100. 陈龙飞, 聂僖曼, 梁沛, 李飞, 韩召军. 2020. 基于转录组数据的害虫抗药性综合检测方法[J]. 植物保护学报, 47(1):18-25.[Chen L F,Nie X M,Liang P,Li F,Han Z J. 2020. The comprehensive method for detecting insecticide resistance by RNA-Seq data analysis[J]. Journal of Plant Protection, 47(1):18-25.]doi: 10.13802/j.cnki.zwbhxb.2020.2019100. 崔丽, 夏长秀, 王立, 王芹芹, 黄伟玲, 袁会珠, 芮昌辉. 2020.成虫玻璃管药膜法测定柑橘木虱田间种群对6种杀虫剂的敏感性[J]. 农药学学报, 22(6):1094-1098.[Cui L,Xia C X, Wang L, Wang Q Q, Huang W L, Yuan H Z,Rui C H. 2020. Sensitivity of field populations of Diaphorina citri(Kuwayama)adults to six insecticides using residual film in glass tubes[J]. Chinese Journal of Pesticide Science,22(6):1094-1098.]doi:10.16801/j.issn. 1008-7303.2020.0135. 邓明学, 潘振兴, 谭有龙, 唐际飞, 覃旭, 陈贵峰, 唐明丽. 2012. 广西果园柑橘木虱对毒死蜱等6种农药的抗药性监测[J]. 中国植保导刊, 47(1):18-25.[Deng M X, Pan Z X,Tan Y L,Tang J F,Qin X,Chen G F,Tang M L. 2012. Resistance monitoring of citrus psyllids to 6 pesticides including chlorpyrifos in Guangxi orchards[J]. China Plant Protection, 47(1):18-25.]doi: 10.3969/j.issn.1672-6820.2012.04.016. 胡双双, 戴素明, 张阳, 郭紫晶, 邓子牛, 李大志. 2022. 柑橘木虱防控药剂筛选及矿物油对其增效作用评价[J]. 植物保护, 48(2):247-254.[Hu S S, Dai S M, Zhang Y, Guo Z J, Deng Z N, Li D Z. 2022. Screening of insecticides for controlling Diaphorina citri and synergism of mineral oil to insecticide[J]. Plant Protection, 48(2):247-254.]doi: 10.16688/j.zwbh.2021018. 胡妍月, 孙杨, 邹志文, 黄水金. 2022. 江西省柑橘木虱对五种常规药剂的抗药性[J]. 应用昆虫学报, 59(2):419-425.[Hu Y Y, Sun Y, Zou Z W, Huang S J. 2022. Resistance of Diaphorina citri to five conventional insecticides in Jiangxi Province[J]. Chinese Journal of Applied Entomology, 59(2):419-425.]doi:10.7679/j.issn.2095-1353. 2022.045. 江宏燕, 吴丰年, 王妍晶, 陶磊, 王吉锋, 刘喆, 岑伊静. 2018.亚洲柑橘木虱的起源、分布和扩散能力研究进展[J]. 环境昆虫学报, 40(5):1014-1020.[Jiang H Y, Wu F N,Wang Y J, Tao L, Wang J F, Liu Z, Cen Y J. 2018. Research advances on the origin,distribution and dispersal of the Asian citrus psyllid(Diaphorina citri Kuwayama)[J]. Journal of Environmental Entomology, 40(5):1014-1020.]doi: 10.3969/j.issn.1674-0858.2018.05.8. 金剑雪. 2017. 贵州白背飞虱抗药性动态监测及其抗性相关基因的转录组学分析[D]. 贵阳:贵州大学.[Jin J X. 2017. Study on dynamic monitoring of insecticide resistance and resistance-related genes based on transcriptome data of Sogatella furcifera(Horváth)in Guizhou Province[D]. Guiyang:Guizhou University.] 刘斌. 2015. 柑橘木虱对杀虫剂敏感性检测及抗药性相关基因分析[D]. 重庆:西南大学.[Liu B. 2015. Detection of insecticides susceptibility and analyses of insecticides resistance-related genes[D]. Chongqing:Southwest University.] 刘守柱, 张蕾蕾, 胥佳睿, 姜晓娇, 郑暖, 于迎鑫. 2021. 亚致死剂量高效氯氰菊酯对莱氏绿僵菌的增效作用[J]. 河南农业科学, 50(5):74-79.[Liu S Z, Zhang L L, Xu J R,Jiang X J, Zheng N, Yu Y X. 2021. Synergetic effects of sublethal dosage of beta-cypermethrin on Metarhizium rileyi against Spodoptera litura[J]. Journal of Henan Agricultural Sciences, 50 (5):74-79.]doi: 10.15933/j.cnki.1004-3268.2021.05.010. 马亿, 张文兵, 张宏玲, 马红悦, 韩海斌, 庞保平, 谭瑶. 2022.牧草盲蝽CYP6A13和钠离子通道LPVSSC基因的克隆及参与高效氯氟氰菊酯的抗性分析[J]. 环境昆虫学报, 44(5):1252-1263.[Ma Y, Zhang W B, Zhang H L, Ma H Y,Han H B,Pang B P,Tan Y. 2022. Cloning of CYP6A13 and sodium channel gene LPVSSC and their involvement in analysis on the resistance of lambda-cyhalothrin in Lygus pratensis(Hemiptera:Miridae)[J]. Journal of Environmental Entomology, 44 (5):1252-1263.]doi: 10.3969/j.issn.1674-0858.2022.05.19. 宋晓兵, 崔一平, 彭埃天, 凌金锋, 陈霞. 2021. 广东肇庆柑橘木虱田间种群对常用药剂的抗药性[J]. 环境昆虫学报, 43(5):1321-1324.[Song X B, Cui Y P, Peng A T, Ling J F, Chen X. 2021. Resistance to commonly used insecticides of Diaphorina citri field populations in Zhaoqing City,Guangdong Province[J]. Journal of Environmental Entomology, 43 (5):1321-1324.]doi: 10.3969/j.issn.1674-0858.2021.05.29. 宋晓兵, 彭埃天, 陈霞, 程保平, 周娟, 凌金锋. 2015. 高效氯氟·噻虫嗪等6种药剂对柑橘木虱的防治效果[J]. 农药, 54(12):915-917.[Song X B, Peng A T, Chen X, Cheng B P, Zhou J, Ling J F. 2015. Control effects of six pesticides such as beta-cypermethrin thiamethoxam on citrus psyllid[J]. Agrochemicals,54(12):915-917.]doi:10. 16820/j.cnki.1006-0413.2015.12.015. 宋晓兵, 彭埃天, 崔一平, 凌金锋, 程保平, 陈霞. 2019. 九里香挥发物对柑橘木虱的引诱效果及混配筛选试验[J]. 植物保护学报, 46(3):589-594.[Song X B, Peng A T, Cui Y P,Ling J F,Cheng B P,Chen X. 2019. Trapping effects of Murraya exotica volatiles on Asian citrus psyllid Diaphorina citri and mixed screening test[J]. Journal of Plant Protection,46(3):589-594.]doi: 10.13802/j.cnki.zwbhxb.2019.2018031. 宋晓兵, 彭埃天, 崔一平, 凌金锋, 程保平, 张炼辉. 2018. 球孢白僵菌诱导的柑橘木虱免疫应答转录组分析[J]. 应用昆虫学报, 55(4):636-645.[Song X B, Peng A T, Cui Y P, Ling J F, Cheng B P, Zhang L H. 2018. Transcriptome analysis of immune responses of Diaphorina citri Kuwayama(Homoptera:Psyllidae)infected with Beauveria bassiana[J]. Chinese Journal of Applied Entomology, 55(4):636-645.]doi: 10.7679/j.issn.2095-1353.2018.080. 孙丽娜, 张怀江, 刘孝贺, 仇贵生. 2020. 基于转录组的苹小卷叶蛾杀虫剂靶标及解毒代谢相关基因分析[J]. 昆虫学报, 63(4):470-481.[Sun L N, Zhang H J, Liu X H, Qiu G S. 2020. Transcriptomic analysis of insecticide target and detoxification related genes in Adoxophyes orana(Lepidoptera:Tortricidae)[J]. Acta Entomologica Sinica, 63 (4):470-481.]doi: 10.16380/j.kcxb.2020.04.010. 田发军, 刘家莉, 曾鑫年. 2018. 柑橘木虱抗药性研究进展[J]. 应用昆虫学报, 55(4):565-573.[Tian F J, Liu J L,Zeng X N. 2018. Progress in research on insecticide resistance in the Asian citrus psyllid, Diaphorina citri[J].Chinese Journal of Applied Entomology,55(4):565-573.]doi:10.7679/j.issn.2095-1353.2018.071.徐鹿, 罗光华, 金瑜剑, 徐德进, 徐广春, 顾中言. 2021. 转录组测序分析毒死蜱与醚菊酯混配对二化螟毒杀的增效机制[J]. 江苏农业学报, 37 (2):317-325.[Xu L, Luo G H,Jin Y J, Xu D J, Xu G C, Gu Z Y. 2021. Study on the synergistic mechanism of chlorpyrifos and ethofenprox mixture in poisoning Chilo suppressalis based on transcriptome sequencing[J]. Jiangsu Journal of Agricultural Sciences, 37(2):317-325.]doi:10.3969/j.issn.1000-4440. 2021.02.006. 詹兴堆. 2019. 不同柑橘木虱种群对6种常用杀虫剂的抗药性测定[J]. 南方农业学报, 50(12):2713-2719.[Zhan X D. 2019. Resistance of different Diaphorina citri populations to six common insecticides[J]. Journal of Southern Agriculture, 50 (12):2713-2719.]doi: 10.3969/j.issn.2095-1191.2019.12.12. 朱江, 邱星辉. 2021. 昆虫抗药性相关细胞色素P450基因的表达调控机制[J]. 昆虫学报, 64(1):109-120.[Zhu J, Qiu X H. 2021. Molecular mechanisms of expression regulation of insect cytochrome P450 genes involved in insecticide resistance[J]. Acta Entomologica Sinica, 64 (1):109-120.]doi: 10.16380/j.kcxb.2021.01.012. Bové J M. 2006. Huanglongbing:A destructive, newly-emerging, century-old disease of citrus[J]. Journal of Plant Pathology, 88 (1):7-37. doi: 10.4454/jpp.v88i1.828.
Cen Y J, Zhang L N, Xia Y L, Guo J, Deng X L, Zhou W J,Sequeira R, Gao J Y, Wang Z R, Yue J Q, Gao Y Q. 2012. Detection of ‘Candidatus Liberibacter asiaticus’ in Cacopsylla(Psylla)citrisuga(Hemiptera:Psyllidae)[J].
Florida Entomologist,95(2):303-309. doi:10.2307/23268549.Feyereisen R. 2011. Insect CYP genes and P450 enzymes[J].Insect Molecular Biology and Biochemistry, 8:263-316.doi:10.1016/B978-0-12-809633-8.04040-1.
Feyereisen R, Dermauw W, Leeuwen T V. 2015. Genotype to phenotype,the molecular and physiological dimensions of resistance in arthropods[J]. Pesticide Biochemistry and Physiology, 121:61-77. doi: 10.1016/j.pestbp.2015.01.004.
Fujii-Kuriyama Y, Mimura J. 2005. Molecular mechanisms of AhR functions in the regulation of cytochrome P450 genes[J]. Biochemical and Biophysical Research Communications, 338 (1):311-317. doi: 10.1016/j.bbrc.2005.08.162.
Halbert S E, Manjunath K L. 2004. Asian citrus psyllids(Sternorrhyncha:Psyllidae)and greening disease of citrus:A literature review and assessment of risk in Florida[J].Florida Entomologist, 87 (3):330-353.
Kanga L H B, Julius E, Muhammad H, Jawwad Q, Philip S. 2016. Monitoring for insecticide resistance in Asian citrus psyllid(Hemiptera:Psyllidae)populations in Florida[J].Journal of Economic Entomology, 109(2):832-836. doi: 10.1093/jee/tov348.
Khan R R, Al-Ghafri T H A, Al-Khatri S A H, Al-Mazidi I S S, Al-Rawahi F G. 2020. Resistance to deltamethrin and fenitrothion in dubas bug,Ommatissus lybicus de Bergevin(Homoptera:Tropiduchidae)and possible bioche-mical mechanisms[J]. Scientific Reports, 10 (1):13220. doi: 10.1038/s41598-020-70150-7.
Killiny N, Hajeri S, Tiwari S, Gowda S, Stelinski L L. 2014.Double-stranded RNA uptake through topical application,mediates silencing of five CYP4 genes and suppresses insecticide resistance in Diaphorina citri[J]. PLoS One, 9(10):e110536. doi: 10.1371/journal.pone.0110536.
Love M I, Huber W, Anders S. 2014. Moderated estimation of fold change and dispersion for RNA-Seq data with DESeq2[J]. Genome Biology, 15(12):550. doi: 10.1186/s13059-014-0550-8.
Oakeshott J G, Home I, Sutherland T D, Russell R J. 2003.The genomics of insecticide resistance[J]. Genome Biology, 4 (1):202. doi: 10.1186/gb-2003-4-1-202.,
Panini M, Manicardi G C, Moores G D, Mazzoni E. 2016. An overview of the main pathways of metabolic resistance in insects[J]. Invertebrate Survival Journal, 13 (9):326-335.
Pierre D W, Melissa H P, Stephen R P. 2015. SNP genotyping and population genomics from expressed sequences-current advances and future possibilities[J]. Molecular Ecology, 24 (10):2310-2323. doi: 10.1111/mec.13165.
Tian F J, Mo X F, Rizvi S A H, Li C F, Zeng X N. 2018. Detection and biochemical characterization of insecticide resistance in field populations of Asian citrus psyllid in Guangdong of China[J]. Scientific Reports, 8(1):12587.doi: 10.1038/s41598-018-30674-5.
Tiwari S, Mann R S, Rogers M E, Stelinski L L. 2011b. Insecticide resistance in field populations of Asian citrus psyllid in Florida[J]. Pest Management Science, 67(10):1258-1268. doi: 10.1002/ps.2181.
Tiwari S, Pelz-Stelinski K, Mann R S, Stelinski L L. 2011a.Glutathione transferase and cytochrome P450 (general oxidase)activity levels in Candidatus Liberibacter asiaticus-infected and uninfected Asian citrus psyllid(Hemiptera:Psyllidae)[J]. Annals of the Entomological Society of America, 104 (2):297-305. doi: 10.1603/AN10128.
Wen G Z. 2017. A simple process of RNA-Sequence analyses by Hisat2,Htseq and DESeq2[C]//Proceedings of the 2017 international conference on biomedical engineering and bioinformatics. New York:Association for Computing Machinery:11-15. doi: 10.1145/3143344.3143354.
Yu F X, Guan K L. 2013. The Hippo pathway:Regulators and regulations[J]. Genes & Development, 27(4):355-371. doi: 10.1101/gad.210773.112.
Yu G G, Wang L G, Han Y Y, He Q Y. 2012. clusterProfiler:An R package for comparing biological themes among gene clusters[J]. OMICS:A Journal of Integrative Biology, 16 (5):284-287. doi: 10.1089/omi.2011.0118.
Zhen C A, Gao X W. 2016. A point mutation(L1015F) of the voltage-sensitive sodium channel gene associated with lambda-cyhalothrin resistance in Apolygus lucorum(Meyer-Dür) population from the transgenic Bt cotton field of China[J]. Pesticide Biochemistry and Physiology, 127:82-89. doi: 10.1016/j.pestbp.2015.09.011.
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