莲草直胸跳甲气味结合蛋白鉴定及其在触角中的表达分析
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摘要
为阐明莲草直胸跳甲Agasicles hygrophila对其寄主植物喜旱莲子草Alternanthera philoxeroides的专一性识别过程中气味结合蛋白(odorant-binding protein,OBP)的作用,基于莲草直胸跳甲三代全长转录组测序结果筛选获得其所有OBP基因并进行生物信息学分析,以最大似然法构建莲草直胸跳甲OBP的系统进化树,并通过定量PCR分析这些OBP基因在其触角中的表达量。结果表明,全长转录组测序结果中共鉴定得到53个莲草直胸跳甲AhygOBP基因,所编码蛋白有36个属于Minus-C家族,11个属于Classic家族,5个属于Plus-C家族,1个属于Atypical家族。进化分析结果显示,大部分莲草直胸跳甲AhygOBP与其它目昆虫OBP为直系同源。莲草直胸跳甲AhygOBP基因在触角中的表达分析结果显示,将表达量最低的AhygOBP31作为基准1,表达量最高的依次为AhygOBP53、AhygOBP32、AhygOBP13、AhygOBP28、AhygOBP44、AhygOBP45、AhygOBP29、AhygOBP37、AhygOBP24,其中AhygOBP53的表达量是AhygOBP31的106 000倍。推测这些触角中高表达的AhygOBP可能在宿主植物识别中发挥重要作用。
Alligator weed flea beetle, Agasicles hygrophila, is a special natural enemy of the global malignant invasive weed Alternanthera philoxeroides. Odorant-binding proteins(OBP) plays an important role in insect host-seeking. To identify the olfactory mechanisms underlying host preference and localization, the OBP genes of A. hygrophila were obtained from the results and analysis of the previous three-generation full-length transcriptome sequencing. Moreover, the expression levels of these OBPs in the antennae were determined by real time quantitative PCR. The results showed that 53 AhygOBP genes were identified in the full-length transcriptome sequencing. Among them, 36 of the encoded proteins belonged to Minus-C family, 11 to Classic family, five to Plus-C family, and one to Atypical family. Phylogenetic analysis revealed that most AhygOBPs were orthologous to other insect OBPs. The lowest expression level of AhygOBP31 was set to 1, AhygOBP53, AhygOBP32, AhygOBP13, AhygOBP28,AhygOBP44, AhygOBP45, AhygOBP29, AhygOBP37, AhygOBP24 had high expression, of which AhygOBP53 was the highest, which was 106 000 times of AhygOBP31. It indicated that high expression of OBPs in antennae might play an important role in host plant recognition.
Alligator weed flea beetle, Agasicles hygrophila, is a special natural enemy of the global malignant invasive weed Alternanthera philoxeroides. Odorant-binding proteins(OBP) plays an important role in insect host-seeking. To identify the olfactory mechanisms underlying host preference and localization, the OBP genes of A. hygrophila were obtained from the results and analysis of the previous three-generation full-length transcriptome sequencing. Moreover, the expression levels of these OBPs in the antennae were determined by real time quantitative PCR. The results showed that 53 AhygOBP genes were identified in the full-length transcriptome sequencing. Among them, 36 of the encoded proteins belonged to Minus-C family, 11 to Classic family, five to Plus-C family, and one to Atypical family. Phylogenetic analysis revealed that most AhygOBPs were orthologous to other insect OBPs. The lowest expression level of AhygOBP31 was set to 1, AhygOBP53, AhygOBP32, AhygOBP13, AhygOBP28,AhygOBP44, AhygOBP45, AhygOBP29, AhygOBP37, AhygOBP24 had high expression, of which AhygOBP53 was the highest, which was 106 000 times of AhygOBP31. It indicated that high expression of OBPs in antennae might play an important role in host plant recognition.
引文
Andersson MN,Grosse-Wilde E,Keeling CI,Bengtsson JM,Yuen MM,Li M,Hillbur Y,Bohlmann J,Hansson BS,Schlyter F.2013.Antennal transcriptome analysis of the chemosensory gene families in the tree killing bark beetles,Ips typographus and Dendroctonus ponderosae(Coleoptera:Curculionidae:Scolytinae).BMC Genomics,14:198
Antony B,Soffan A,Jakse J,Abdelazim MM,Aldosari SA,Aldawood AS,Pain A.2016.Identification of the genes involved in odorant reception and detection in the palm weevil Rhynchophorus ferrugineus,an important quarantine pest,by antennal transcriptome analysis.BMC Genomics,17:69
Benton R,Vannice KS,Gomez-Diaz C,Vosshall LB.2009.Variant ionotropic glutamate receptors as chemosensory receptors in Drosophila.Cell,136(1):149-162
Buckingham GR.1996.Biological control of alligatorweed,Alternanthera philoxeroides,the world’s first aquatic weed success story.Castanea,61(3):232-243
Deng S,Yin J,Zhong T,Cao Y,Li K.2012.Function and immunocytochemical localization of two novel odorant-binding proteins in olfactory sensilla of the scarab beetle Holotrichia oblita Faldermann(Coleoptera:Scarabaeidae).Chemical Senses,37(2):141-150
Dippel S,Oberhofer G,Kahnt J,Gerischer L,Opitz L,Schachtner J,Stanke M,Schutz S,Wimmer EA,Angeli S.2014.Tissue-specific transcriptomics,chromosomal localization,phylogeny of chemosensory and odorant binding proteins from the red flour beetle Tribolium castaneum reveal subgroup specificities for olfaction or more general functions.BMC Genomics,15:1141
Gu XC,Zhang YN,Kang K,Dong SL,Zhang LW.2015.Antennal transcriptome analysis of odorant reception genes in the red turpentine beetle(RTB),Dendroctonus valen.PLoS ONE,10(5):e0125159
Hallem EA,Dahanukar A,Carlson JR.2006.Insect odor and taste receptors.Annual Review of Entomology,51:113-135
Hu P,Wang J,Cui M,Tao J,Luo Y.2016.Antennal transcriptome analysis of the Asian longhorned beetle Anoplophora glabripenni.Scientific Reports,6:26652
Ji SS,Zhuang XL,Liu NY.2018.Identification and expression profiling analysis of odorant binding protein genes from Xylotrechus quadripes Chevrolat(Coleoptera:Cerambycidae).Journal of Sichuan Agricultural University,36(2):193-202(in Chinese)[吉帅帅,庄翔麟,刘乃勇.2018.灭字脊虎天牛触角转录组中气味结合蛋白基因的鉴定及表达谱分析.四川农业大学学报,36(2):193-202]
Jia D,Wang YX,Liu YH,Hu J,Guo YQ,Gao LL,Ma RY.2018.SMRT sequencing of full-length transcriptome of flea beetle Agasicles hygrophila(Selman and Vogt).Scientific Reports,8(1):2197
Ju Q,Li X,Jiang XJ,Qu MJ,Guo XQ,Han ZJ,Li F.2014.Transcriptome and tissue-specific expression analysis of OBP and CSPgenes in the dark black chafer.Archives of Insect Biochemistry and Physiology,87(4):177-200
Julien MH,Stanley JN.1999.The management of alligator weed,a challenge for the new millennium.//Practical weed management:protecting agriculture and the environment:Proceedings of the10th Biennial Noxious Weeds Conference.Ballina:New South Wales Department of Agriculture Agriculture,pp.2-13
Laughlin JD,Ha TS,Jones DNM,Smith DP.2008.Activation of pheromone-sensitive neurons is mediated by conformational activation of pheromone-binding protein.Cell,133(7):1255-1265
Li GW,Zhang Y,Li YP,Wu JX,Xu XL.2016.Cloning,expression,and functional analysis of three odorant-binding proteins of the oriental fruit moth,Grapholita molesta(Busck)(Lepidoptera:Tortricidae).Archives of Insect Biochemistry and Physiology,91(2):67-87
Li L,Zhou YT,Tan Y,Zhou XR,Pang BP.2017.Identification of odorant-binding protein genes in Galeruca daurica(Coleoptera:Chrysomelidae)and analysis of their expression profiles.Bulletin of Entomological Research,107(4):550-561
Li XM,Zhu XY,Wang ZQ,Wang Y,He P,Chen G,Sun L,Deng DG,Zhang YN.2015.Candidate chemosensory genes identified in Colaphellus bowringi by antennal transcriptome analysis.BMCGenomics,16:1028
Li ZY,Xie Y.2002.Invasive alien species in China.Beijing:Forestry Publishing Company of China(in Chinese)[李振宇,解焱.2002.中国外来入侵种.北京:中国林业出版社]
Lin KJ,Wu KM,Zhang YJ,Guo YY.2008.The feeding and oviposition behaviors of Bemisia tabaci(Gennadius)biotype B on five host plants.Journal of Plant Protection,35(3):199-204(in Chinese)[林克剑,吴孔明,张永军,郭予元.2008.B型烟粉虱成虫对五种寄主植物的取食和产卵行为.植物保护学报,35(3):199-204]
Liu NY,Li ZB,Zhao N,Song QS,Zhu JY,Yang B.2018.Identification and characterization of chemosensory gene families in the bark beetle,Tomicus yunnanensis.Comparative Biochemistry and Physiology Part D:Genomics&Proteomics,25:73-85
Ma RY,Wang Ren.2005.Invasive mechanism and biological control of alligatorweed,Alternanthera philoxeroides(Amaranthaceae),in China.Chinese Journal of Applied and Environmental Biology,11(2):246-250(in Chinese)[马瑞燕,王韧.2005.喜旱莲子草在中国的入侵机理及其生物防治.应用与环境生物学报,11(2):246-250]
Mao Y,Xu X,Xu W,Ishida Y,Leal WS,Ames JB,Clardy J.2010.Crystal and solution structures of an odorant-binding protein from the southern house mosquito complexed with an oviposition pheromone.Proceedings of the National Academy of Sciences of the United States of America,107(44):19102-19107
McCormick AC,Unsicker SB,Gershenzon J.2012.The specificity of herbivore-induced plant volatiles in attracting herbivore enemies.Trends in Plant Science,17(5):303-310
Pan XY,Geng YP,Alejandro SOSA,Zhang WJ,Li B,Chen JK.2007.Invasive Alternanthera philoxeroides:biology,ecology and management.Acta Phytotaxonomica Sinica,45(6):884-900(in Chinese)[潘晓云,耿宇鹏,Alejandro SOSA,张文驹,李博,陈家宽.2007.入侵植物喜旱莲子草--生物学、生态学及管理.植物分类学报,45(6):884-900]
Pelosi P,Maida R.1995.Odorant-binding proteins in insects.Comparative Biochemistry and Physiology Part B:Biochemistry&Molecular Biology,111(3):503-514
Pelosi P,Iovinella I,Felicioli A,Dani FR.2014.Soluble proteins of chemical communication:an overview across arthropods.Frontiers in Physiology,5:320
Rebijith KB,Asokan R,Hande HR,Kumar NK,Krishna V,Vinutha J,Bakthavatsalam N.2016.RNA interference of odorant-binding protein 2(OBP2)of the cotton aphid,Aphis gossypii(Glover),resulted in altered electrophysiological responses.Applied Biochemistry and Biotechnology,178(2):251-266
Rybczynski R,Vogt RG,Lerner MR.1990.Antennal-specific pheromone-degrading aldehyde oxidases from the moths Antheraea polyphemus and Bombyx mori.The Journal of Biological Chemistry,265(32):19712-19715
Sun L,Gu SH,Xiao HJ,Zhou JJ,Guo YY,Liu ZW,Zhang YJ.2013.The preferential binding of a sensory organ specific odorant binding protein of the alfalfa plant bug Adelphocoris lineolatus AlinOBP10 to biologically active host plant volatiles.Journal of Chemical Ecology,39(9):1221-1231
Tian L,Zhang Y,Zhao YF.2015.The next generation sequencing technology and its applications.Biotechnology Bulletin,31(11):1-8(in Chinese)[田李,张颖,赵云峰.2015.新一代测序技术的发展和应用.生物技术通报,31(11):1-8]
Wan X,Qian K,Du Y.2015.Synthetic pheromones and plant volatiles alter the expression of chemosensory genes in Spodoptera exigua.Scientific Reports,5:17320
Wang J,Li DZ,Min SF,Mi F,Zhou SS,Wang MQ.2014.Analysis of chemosensory gene families in the beetle Monochamus alternatus and its parasitoid Dastarcus helophoroides.Comparative Biochemistry and Physiology Part D:Genomics&Proteomics,11:1-8
Wang R,Zhang XM,Li FQ,Wu F,Li HL,Luo C.2016.Cloning and prokaryotic expression of odorant binding protein OBP8 in MED cryptic species Bemisia tabaci and the binding characteristics with plant volatiles.Journal of Plant Protection,43(1):32-39(in Chinese)[王然,张晓曼,李峰奇,吴帆,李红亮,罗晨.2016.烟粉虱MED隐种气味结合蛋白OBP8的克隆、原核表达及与植物挥发物的结合特性.植物保护学报,43(1):32-39]
Wang YL,Zhang J,Chen Q,Zhao HB,Wang JT,Wen M,Xi JH,Ren BZ.2018.Identification and evolution of olfactory genes in the small poplar longhorn beetle Saperda populnea.Comparative Biochemistry and Physiology Part D:Genomics&Proteomics,26:58-68
Yin J,Wang CQ,Fang CQ,Zhang S,Cao YZ,Li KB,Leal WS.2019.Functional characterization of odorant-binding proteins from the scarab beetle Holotrichia oblita based on semiochemical-induced expression alteration and gene silencing.Insect Biochemistry and Molecular Biology,104:11-19
Zhang B,Zhang W,Nie RE,Li WZ,Segraves KA,Yang XK,Xue HJ.2016.Comparative transcriptome analysis of chemosensory genes in two sister leaf beetles provides insights into chemosensory speciation.Insect Biochemistry and Molecular Biology,79:108-118
Zhou JJ.2010.Odorant-binding proteins in insects.Vitamins and Hormones,83:241-272
Antony B,Soffan A,Jakse J,Abdelazim MM,Aldosari SA,Aldawood AS,Pain A.2016.Identification of the genes involved in odorant reception and detection in the palm weevil Rhynchophorus ferrugineus,an important quarantine pest,by antennal transcriptome analysis.BMC Genomics,17:69
Benton R,Vannice KS,Gomez-Diaz C,Vosshall LB.2009.Variant ionotropic glutamate receptors as chemosensory receptors in Drosophila.Cell,136(1):149-162
Buckingham GR.1996.Biological control of alligatorweed,Alternanthera philoxeroides,the world’s first aquatic weed success story.Castanea,61(3):232-243
Deng S,Yin J,Zhong T,Cao Y,Li K.2012.Function and immunocytochemical localization of two novel odorant-binding proteins in olfactory sensilla of the scarab beetle Holotrichia oblita Faldermann(Coleoptera:Scarabaeidae).Chemical Senses,37(2):141-150
Dippel S,Oberhofer G,Kahnt J,Gerischer L,Opitz L,Schachtner J,Stanke M,Schutz S,Wimmer EA,Angeli S.2014.Tissue-specific transcriptomics,chromosomal localization,phylogeny of chemosensory and odorant binding proteins from the red flour beetle Tribolium castaneum reveal subgroup specificities for olfaction or more general functions.BMC Genomics,15:1141
Gu XC,Zhang YN,Kang K,Dong SL,Zhang LW.2015.Antennal transcriptome analysis of odorant reception genes in the red turpentine beetle(RTB),Dendroctonus valen.PLoS ONE,10(5):e0125159
Hallem EA,Dahanukar A,Carlson JR.2006.Insect odor and taste receptors.Annual Review of Entomology,51:113-135
Hu P,Wang J,Cui M,Tao J,Luo Y.2016.Antennal transcriptome analysis of the Asian longhorned beetle Anoplophora glabripenni.Scientific Reports,6:26652
Ji SS,Zhuang XL,Liu NY.2018.Identification and expression profiling analysis of odorant binding protein genes from Xylotrechus quadripes Chevrolat(Coleoptera:Cerambycidae).Journal of Sichuan Agricultural University,36(2):193-202(in Chinese)[吉帅帅,庄翔麟,刘乃勇.2018.灭字脊虎天牛触角转录组中气味结合蛋白基因的鉴定及表达谱分析.四川农业大学学报,36(2):193-202]
Jia D,Wang YX,Liu YH,Hu J,Guo YQ,Gao LL,Ma RY.2018.SMRT sequencing of full-length transcriptome of flea beetle Agasicles hygrophila(Selman and Vogt).Scientific Reports,8(1):2197
Ju Q,Li X,Jiang XJ,Qu MJ,Guo XQ,Han ZJ,Li F.2014.Transcriptome and tissue-specific expression analysis of OBP and CSPgenes in the dark black chafer.Archives of Insect Biochemistry and Physiology,87(4):177-200
Julien MH,Stanley JN.1999.The management of alligator weed,a challenge for the new millennium.//Practical weed management:protecting agriculture and the environment:Proceedings of the10th Biennial Noxious Weeds Conference.Ballina:New South Wales Department of Agriculture Agriculture,pp.2-13
Laughlin JD,Ha TS,Jones DNM,Smith DP.2008.Activation of pheromone-sensitive neurons is mediated by conformational activation of pheromone-binding protein.Cell,133(7):1255-1265
Li GW,Zhang Y,Li YP,Wu JX,Xu XL.2016.Cloning,expression,and functional analysis of three odorant-binding proteins of the oriental fruit moth,Grapholita molesta(Busck)(Lepidoptera:Tortricidae).Archives of Insect Biochemistry and Physiology,91(2):67-87
Li L,Zhou YT,Tan Y,Zhou XR,Pang BP.2017.Identification of odorant-binding protein genes in Galeruca daurica(Coleoptera:Chrysomelidae)and analysis of their expression profiles.Bulletin of Entomological Research,107(4):550-561
Li XM,Zhu XY,Wang ZQ,Wang Y,He P,Chen G,Sun L,Deng DG,Zhang YN.2015.Candidate chemosensory genes identified in Colaphellus bowringi by antennal transcriptome analysis.BMCGenomics,16:1028
Li ZY,Xie Y.2002.Invasive alien species in China.Beijing:Forestry Publishing Company of China(in Chinese)[李振宇,解焱.2002.中国外来入侵种.北京:中国林业出版社]
Lin KJ,Wu KM,Zhang YJ,Guo YY.2008.The feeding and oviposition behaviors of Bemisia tabaci(Gennadius)biotype B on five host plants.Journal of Plant Protection,35(3):199-204(in Chinese)[林克剑,吴孔明,张永军,郭予元.2008.B型烟粉虱成虫对五种寄主植物的取食和产卵行为.植物保护学报,35(3):199-204]
Liu NY,Li ZB,Zhao N,Song QS,Zhu JY,Yang B.2018.Identification and characterization of chemosensory gene families in the bark beetle,Tomicus yunnanensis.Comparative Biochemistry and Physiology Part D:Genomics&Proteomics,25:73-85
Ma RY,Wang Ren.2005.Invasive mechanism and biological control of alligatorweed,Alternanthera philoxeroides(Amaranthaceae),in China.Chinese Journal of Applied and Environmental Biology,11(2):246-250(in Chinese)[马瑞燕,王韧.2005.喜旱莲子草在中国的入侵机理及其生物防治.应用与环境生物学报,11(2):246-250]
Mao Y,Xu X,Xu W,Ishida Y,Leal WS,Ames JB,Clardy J.2010.Crystal and solution structures of an odorant-binding protein from the southern house mosquito complexed with an oviposition pheromone.Proceedings of the National Academy of Sciences of the United States of America,107(44):19102-19107
McCormick AC,Unsicker SB,Gershenzon J.2012.The specificity of herbivore-induced plant volatiles in attracting herbivore enemies.Trends in Plant Science,17(5):303-310
Pan XY,Geng YP,Alejandro SOSA,Zhang WJ,Li B,Chen JK.2007.Invasive Alternanthera philoxeroides:biology,ecology and management.Acta Phytotaxonomica Sinica,45(6):884-900(in Chinese)[潘晓云,耿宇鹏,Alejandro SOSA,张文驹,李博,陈家宽.2007.入侵植物喜旱莲子草--生物学、生态学及管理.植物分类学报,45(6):884-900]
Pelosi P,Maida R.1995.Odorant-binding proteins in insects.Comparative Biochemistry and Physiology Part B:Biochemistry&Molecular Biology,111(3):503-514
Pelosi P,Iovinella I,Felicioli A,Dani FR.2014.Soluble proteins of chemical communication:an overview across arthropods.Frontiers in Physiology,5:320
Rebijith KB,Asokan R,Hande HR,Kumar NK,Krishna V,Vinutha J,Bakthavatsalam N.2016.RNA interference of odorant-binding protein 2(OBP2)of the cotton aphid,Aphis gossypii(Glover),resulted in altered electrophysiological responses.Applied Biochemistry and Biotechnology,178(2):251-266
Rybczynski R,Vogt RG,Lerner MR.1990.Antennal-specific pheromone-degrading aldehyde oxidases from the moths Antheraea polyphemus and Bombyx mori.The Journal of Biological Chemistry,265(32):19712-19715
Sun L,Gu SH,Xiao HJ,Zhou JJ,Guo YY,Liu ZW,Zhang YJ.2013.The preferential binding of a sensory organ specific odorant binding protein of the alfalfa plant bug Adelphocoris lineolatus AlinOBP10 to biologically active host plant volatiles.Journal of Chemical Ecology,39(9):1221-1231
Tian L,Zhang Y,Zhao YF.2015.The next generation sequencing technology and its applications.Biotechnology Bulletin,31(11):1-8(in Chinese)[田李,张颖,赵云峰.2015.新一代测序技术的发展和应用.生物技术通报,31(11):1-8]
Wan X,Qian K,Du Y.2015.Synthetic pheromones and plant volatiles alter the expression of chemosensory genes in Spodoptera exigua.Scientific Reports,5:17320
Wang J,Li DZ,Min SF,Mi F,Zhou SS,Wang MQ.2014.Analysis of chemosensory gene families in the beetle Monochamus alternatus and its parasitoid Dastarcus helophoroides.Comparative Biochemistry and Physiology Part D:Genomics&Proteomics,11:1-8
Wang R,Zhang XM,Li FQ,Wu F,Li HL,Luo C.2016.Cloning and prokaryotic expression of odorant binding protein OBP8 in MED cryptic species Bemisia tabaci and the binding characteristics with plant volatiles.Journal of Plant Protection,43(1):32-39(in Chinese)[王然,张晓曼,李峰奇,吴帆,李红亮,罗晨.2016.烟粉虱MED隐种气味结合蛋白OBP8的克隆、原核表达及与植物挥发物的结合特性.植物保护学报,43(1):32-39]
Wang YL,Zhang J,Chen Q,Zhao HB,Wang JT,Wen M,Xi JH,Ren BZ.2018.Identification and evolution of olfactory genes in the small poplar longhorn beetle Saperda populnea.Comparative Biochemistry and Physiology Part D:Genomics&Proteomics,26:58-68
Yin J,Wang CQ,Fang CQ,Zhang S,Cao YZ,Li KB,Leal WS.2019.Functional characterization of odorant-binding proteins from the scarab beetle Holotrichia oblita based on semiochemical-induced expression alteration and gene silencing.Insect Biochemistry and Molecular Biology,104:11-19
Zhang B,Zhang W,Nie RE,Li WZ,Segraves KA,Yang XK,Xue HJ.2016.Comparative transcriptome analysis of chemosensory genes in two sister leaf beetles provides insights into chemosensory speciation.Insect Biochemistry and Molecular Biology,79:108-118
Zhou JJ.2010.Odorant-binding proteins in insects.Vitamins and Hormones,83:241-272