点蜂缘蝽触角转录组及化学感受相关基因的分析
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摘要
【目的】点蜂缘蝽Riptortus pedestris主要为害豆科作物,成虫和若虫刺吸汁液,造成严重损失,目前对该虫的研究较少,基因信息缺乏。本研究旨在获得点蜂缘蝽的触角转录组数据,开发基于嗅觉防治害虫的新方法。【方法】利用Illumina Hi SeqTM4000高通量测序技术测定了点蜂缘蝽成虫触角转录组,并进行了生物学信息分析。【结果】共获得45 802 812条clean reads,包括6.87 Gb(Gen Bank登录号:SRR4429103)。拼接92 259条unigenes,平均长度为618 bp,N50为1 013 bp。在7大数据库中注释到21 365条unigenes。通过进一步转录组数据分析,我们鉴定出219个点蜂缘蝽化学感受相关基因,包括188个嗅觉受体、6个味觉受体、2个离子型受体、4个感觉神经元膜蛋白、8个气味结合蛋白和11个化学感受蛋白。氨基酸序列分析发现,RpedOBP1和RpedOBP2在第6个保守的半胱氨酸后又多了3个保守的半胱氨酸位点,属于加Plus-C气味结合蛋白家族。【结论】本研究获得了点蜂缘蝽触角转录组数据,并鉴定出嗅觉相关基因,结果为今后利用嗅觉基因靶标防治点蜂缘蝽提供了重要的分子生物学信息数据。
【Aim】Riptortus pedestris is a major soybean pest which feeds juice in the stages of adult and nymph,causing serious damage to production.However,R.pedestris has been rarely researched,that leads to the lack of genomic resources.The objective of this study is to obtain the antennal transcriptome data of R.pedestris and to seek new methods for pest control through olfaction.【Methods】The adult antennal transcriptome of R.pedestris is sequenced using Illumina Hi SeqTM4000 platform and analyzed bioinformatically.【Results】In total,45 802 812 clean reads with 6.87 Gb were obtained(GenBank accession no.:SRR4429103).There were 92 259 unigenes with the mean length of 618 bp and an N50 of 1 013 bp.A total of 21 365 unigenes were annotated based on seven databases.By further analyzing the transcriptome data,we identified 219 chemoreception-related genes including 188 for olfactory receptors(ORs),6 for gustatory receptors(GRs),2 for ionotropic receptors(IRs),4 for sensory neuron membrane proteins(SNMPs),8 for odorant-binding proteins(OBPs) and 11 for chemosensory proteins(CSPs).The analysis of amino acid sequence indicated that RpedOBP1 and RpedOBP2 have three additional conserved cysteine residues immediately after the sixth cysteine and belong to the plus-C OBP family.【Conclusion 】 This study acquired the antennal transcriptom data of R.pedestris andidentified olfaction-related genes.The results provide the important information data of molecular biology for the control of R.pedestris using olfaction-related gene targets.
【Aim】Riptortus pedestris is a major soybean pest which feeds juice in the stages of adult and nymph,causing serious damage to production.However,R.pedestris has been rarely researched,that leads to the lack of genomic resources.The objective of this study is to obtain the antennal transcriptome data of R.pedestris and to seek new methods for pest control through olfaction.【Methods】The adult antennal transcriptome of R.pedestris is sequenced using Illumina Hi SeqTM4000 platform and analyzed bioinformatically.【Results】In total,45 802 812 clean reads with 6.87 Gb were obtained(GenBank accession no.:SRR4429103).There were 92 259 unigenes with the mean length of 618 bp and an N50 of 1 013 bp.A total of 21 365 unigenes were annotated based on seven databases.By further analyzing the transcriptome data,we identified 219 chemoreception-related genes including 188 for olfactory receptors(ORs),6 for gustatory receptors(GRs),2 for ionotropic receptors(IRs),4 for sensory neuron membrane proteins(SNMPs),8 for odorant-binding proteins(OBPs) and 11 for chemosensory proteins(CSPs).The analysis of amino acid sequence indicated that RpedOBP1 and RpedOBP2 have three additional conserved cysteine residues immediately after the sixth cysteine and belong to the plus-C OBP family.【Conclusion 】 This study acquired the antennal transcriptom data of R.pedestris andidentified olfaction-related genes.The results provide the important information data of molecular biology for the control of R.pedestris using olfaction-related gene targets.
引文
An XK,Sun L,Liu HW,Liu DF,Ding YX,Li LM,Zhang YJ,Guo YY,2016.Identification and expression analysis of an olfactory receptor gene family in green plant bug Apolygus lucorum(MeyerDür).Sci.Rep.,6:37870.
Becher PG,Flick G,Rozpedowska E,Schmidt A,Hagman A,Lebreton S,Larsson MC,Hansson BS,Pi2kur J,Witzgall P,Bengtsson M,2012.Yeast,not fruit volatiles mediate Drosophila melanogaster attraction,oviposition and development.Funct.Ecol.,26(4):822-828.
Benton R,Vannice KS,Gomez-Diaz C,Vosshall LB,2009.Variant ionotropic glutamate receptors as chemosensory receptors in Drosophila.Cell,136:149-162.
Benton R,Vannice KS,Vosshall LB,2007.An essential role for a CD36-related receptor in pheromone detection in Drosophila.Nature,450:289-293.
Birkett MA,Agelopoulos N,Jensen KMV,Jespersen JB,Pickett JA,Prijs HJ,Thomas G,Trapman JJ,Wadhams LJ,Woodcock CM,2004.The role of volatile semiochemicals in mediating host location and selection by nuisance and disease-transmitting cattle flies.Med.Vet.Entomol.,18(4):313-322.
Clyne PJ,Warr CG,Carlson JR,2000.Candidate taste receptors in Drosophila.Science,287:1830-1834.
Clyne PJ,Warr CG,Freeman MR,Lessing D,Kim J,Carlson JR,1999.A novel family of divergent seven transmembrane proteins:candidate odorant receptors in Drosophila.Neuron,22:327-338.
Gong DP,Zhang HJ,Zhao P,Xia QY,Xiang ZH,2009.The odorant binding protein gene family from the genome of silkworm,Bombyx mori.BMC Genomics,10:332.
Gu SH,Wang SP,Zhang XY,Wu KM,Guo YY,Zhou JJ,Zhang YJ,2011.Identification and tissue distribution of odorant binding protein genes in the lucerne plant bug Adelphocoris lineolatus(Goeze).Insect Biochem.Molec.Biol.,41(4):254-263.
Ha KS,Heo NK,Kim JR,Kim SY,Song SH,1998.Effect of different seeding times and soybean varieties on damage and occurrences of hemiptera insects.RDA J.Crop Prot.,40:32-36.
Jacquin-Joly E,Vogt RG,Fran9ois MC,Nagnan-Le Meillour P,2001.Functional and expression pattern analysis of chemosensory proteins expressed in antennae and pheromonal gland of Mamestra brassicae.Chem.Senses,26:833-844.
Jin YL,Cong B,Wang LY,Zhang HY,Dong H,2015.An analysis of the transcriptome of Epacromius coerulipes(Orthoptera:Acrididae).Acta Entomol.Sin.,58(8):817-825.[金永玲,丛斌,王丽艳,张海燕,董辉,2015.大垫尖翅蝗转录组分析.昆虫学报,58(8):817-825]
Kikuhara Y,2005.The Japanese species of the genus Riptortus(Heteroptera,Alydidae)with description of a new species.Jpn.J.Syst.Entomol.,11:299-311.
Kurtovic A,Widmer A,Dickson B,2007.A single class of olfactory neurons mediates behavioural responses to a Drosophila sex pheromone.Nature,466(7153):542-546.
Larkin M,Blackshields G,Brown N,Chenna R,Mc Gettigan PA,Mc William H,Valentin F,Wallace IM,Wilm A,Lopez R,2007.Clustal W and Clustal X version 2.0.Bioinformatics,23:2947-2948.
Leal WS,2013.Odorant reception in insects:roles of receptors,binding proteins,and degrading enzymes.Annu.Rev.Entomol.,58:373-391.
Mc Kenna MP,Hekmat-Scafe DS,Gaines P,Carlson JR,1994.Putative Drosophila pheromone-binding proteins expressed in a subregion of the olfactory system.J.Biol.Chem.,269:16340-16347.
Meng X,Hu JJ,Liu H,Ouyang GC,Guo MF,2016.Analysis of the transcriptome and olfaction-related genes of Conopomorpha sinensis Bradley(Lepidoptera:Gracilariidae).Acta Entomol.Sin.,59(8):823-830.[孟翔,胡俊杰,刘慧,欧阳革成,郭明昉,2016.荔枝蒂蛀虫转录组及嗅觉相关基因分析.昆虫学报,59(8):823-830]
Ozaki M,Wada-Katsumata A,Fujikawa K,Iwasaki M,Yokohari F,Satoji Y,Nisimura T,Yamaoka R,2005.Ant nestmate and nonnestmate discrimination by a chemosensory sensillum.Science,309:311-314.
Pelosi P,Iovinella I,Felicioli A,Dani FR,2014.Soluble proteins of chemical communication:an overview across arthropods.Front.Physiol.,5:320.
Pikielny C,Hasan G,Rouyer F,Rosbash M,1994.Members of a family of Drosophila putative odorant-binding proteins are expressed in different subsets of olfactory hairs.Neuron,12:35-49.
Plettner E,2002.Insect pheromone olfaction:new targets for the design of species-selective pest control agents.Curr.Med.Chem.,9(10):1075-1085.
Schaefer CW,Panizzi AR,2000.Heteroptera of Economic Importance.CRC Press,Florida.856 pp.
Son CK,Hwang SG,Hyun Y,Choi BS,2000.Field occurrence of stink bug and its damage in soybean.Korean J.Crop Sci.,45:405-410.
Sun HZ,Song YQ,Du J,Wang XD,Cheng ZJ,2016.Identification and tissue distribution of chemosensory protein and odorant binding protein genes in Athetis dissimilis(Lepidoptera:Noctuidae).Appl.Entomol.Zool.,51(3):409-420.
Tomokuni M,Yasunaga T,Takai M,Yamashita I,Kawamura M,Kawasawa T,1993.A Field Guide to Japanese Bugs:Terrestrial Heteropterans.Zenkoku Noson Kyoiku Kyokai,Tokyo.380 pp.
Vogt RG,Riddiford LM,1981.Pheromone binding and inactivation by moth antennae.Nature,293:161-163.
Vogt RG,Riddiford LM,Prestwich GD,1985.Kinetic properties of a sex pheromone-degrading enzyme:the sensillar esterase of Antheraea polyphemus.Proc.Natl.Acad.Sci.USA,82:8827-8831.
Vosshall LB,Stocker RF,2007.Molecular architecture of smell and taste in Drosophila.Annu.Rev.Neurosci.,30:505-533.
Xiao Y,Sun L,Ma XY,Dong K,Liu HW,Wang Q,Guo YY,Liu ZW,Zhang YJ,2016.Identification and characterization of the distinct expression profiles of candidate chemosensory membrane proteins in the antennal transcriptome of Adelphocoris lineolatus(Goeze).Insect Mol.Biol.,26(1):74-91.
Yuan HB,Ding YX,Gu SH,Sun L,Zhu XQ,Liu HW,Dhiloo KH,Zhang YJ,Guo YY,2015,Molecular characterization and expression profiling of odorant-binding proteins in Apolygus lucorum.PLo S ONE,10(10):e0140562.
Zhang QL,Yuan ML,2013.Progress in insect transcriptomics based on the next-generation sequencing technique.Acta Entomol.Sin.,56(12):1489-1508.[张棋麟,袁明龙,2013.基于新一代测序技术的昆虫转录组学研究进展.昆虫学报,56(12):1489-1508]
Zhang YN,Zhu XY,Zhang Q,Yin CY,Dong ZP,Zuo LH,Deng DG,Sun L,Li XM,2016.De novo assembly and characterization of antennal transcriptome reveal chemosensory system in Nysius ericae.J.Asia-Pac.Entomol.,19:1077-1087.
Zhou SH,Zhang J,Zhang SG,Zhang L,2008.Expression of chemosensory proteins in hairs on wings of Locusta migratoria(Orthoptera:Acrididae).J.Appl.Entomol.,132:439-450.
Becher PG,Flick G,Rozpedowska E,Schmidt A,Hagman A,Lebreton S,Larsson MC,Hansson BS,Pi2kur J,Witzgall P,Bengtsson M,2012.Yeast,not fruit volatiles mediate Drosophila melanogaster attraction,oviposition and development.Funct.Ecol.,26(4):822-828.
Benton R,Vannice KS,Gomez-Diaz C,Vosshall LB,2009.Variant ionotropic glutamate receptors as chemosensory receptors in Drosophila.Cell,136:149-162.
Benton R,Vannice KS,Vosshall LB,2007.An essential role for a CD36-related receptor in pheromone detection in Drosophila.Nature,450:289-293.
Birkett MA,Agelopoulos N,Jensen KMV,Jespersen JB,Pickett JA,Prijs HJ,Thomas G,Trapman JJ,Wadhams LJ,Woodcock CM,2004.The role of volatile semiochemicals in mediating host location and selection by nuisance and disease-transmitting cattle flies.Med.Vet.Entomol.,18(4):313-322.
Clyne PJ,Warr CG,Carlson JR,2000.Candidate taste receptors in Drosophila.Science,287:1830-1834.
Clyne PJ,Warr CG,Freeman MR,Lessing D,Kim J,Carlson JR,1999.A novel family of divergent seven transmembrane proteins:candidate odorant receptors in Drosophila.Neuron,22:327-338.
Gong DP,Zhang HJ,Zhao P,Xia QY,Xiang ZH,2009.The odorant binding protein gene family from the genome of silkworm,Bombyx mori.BMC Genomics,10:332.
Gu SH,Wang SP,Zhang XY,Wu KM,Guo YY,Zhou JJ,Zhang YJ,2011.Identification and tissue distribution of odorant binding protein genes in the lucerne plant bug Adelphocoris lineolatus(Goeze).Insect Biochem.Molec.Biol.,41(4):254-263.
Ha KS,Heo NK,Kim JR,Kim SY,Song SH,1998.Effect of different seeding times and soybean varieties on damage and occurrences of hemiptera insects.RDA J.Crop Prot.,40:32-36.
Jacquin-Joly E,Vogt RG,Fran9ois MC,Nagnan-Le Meillour P,2001.Functional and expression pattern analysis of chemosensory proteins expressed in antennae and pheromonal gland of Mamestra brassicae.Chem.Senses,26:833-844.
Jin YL,Cong B,Wang LY,Zhang HY,Dong H,2015.An analysis of the transcriptome of Epacromius coerulipes(Orthoptera:Acrididae).Acta Entomol.Sin.,58(8):817-825.[金永玲,丛斌,王丽艳,张海燕,董辉,2015.大垫尖翅蝗转录组分析.昆虫学报,58(8):817-825]
Kikuhara Y,2005.The Japanese species of the genus Riptortus(Heteroptera,Alydidae)with description of a new species.Jpn.J.Syst.Entomol.,11:299-311.
Kurtovic A,Widmer A,Dickson B,2007.A single class of olfactory neurons mediates behavioural responses to a Drosophila sex pheromone.Nature,466(7153):542-546.
Larkin M,Blackshields G,Brown N,Chenna R,Mc Gettigan PA,Mc William H,Valentin F,Wallace IM,Wilm A,Lopez R,2007.Clustal W and Clustal X version 2.0.Bioinformatics,23:2947-2948.
Leal WS,2013.Odorant reception in insects:roles of receptors,binding proteins,and degrading enzymes.Annu.Rev.Entomol.,58:373-391.
Mc Kenna MP,Hekmat-Scafe DS,Gaines P,Carlson JR,1994.Putative Drosophila pheromone-binding proteins expressed in a subregion of the olfactory system.J.Biol.Chem.,269:16340-16347.
Meng X,Hu JJ,Liu H,Ouyang GC,Guo MF,2016.Analysis of the transcriptome and olfaction-related genes of Conopomorpha sinensis Bradley(Lepidoptera:Gracilariidae).Acta Entomol.Sin.,59(8):823-830.[孟翔,胡俊杰,刘慧,欧阳革成,郭明昉,2016.荔枝蒂蛀虫转录组及嗅觉相关基因分析.昆虫学报,59(8):823-830]
Ozaki M,Wada-Katsumata A,Fujikawa K,Iwasaki M,Yokohari F,Satoji Y,Nisimura T,Yamaoka R,2005.Ant nestmate and nonnestmate discrimination by a chemosensory sensillum.Science,309:311-314.
Pelosi P,Iovinella I,Felicioli A,Dani FR,2014.Soluble proteins of chemical communication:an overview across arthropods.Front.Physiol.,5:320.
Pikielny C,Hasan G,Rouyer F,Rosbash M,1994.Members of a family of Drosophila putative odorant-binding proteins are expressed in different subsets of olfactory hairs.Neuron,12:35-49.
Plettner E,2002.Insect pheromone olfaction:new targets for the design of species-selective pest control agents.Curr.Med.Chem.,9(10):1075-1085.
Schaefer CW,Panizzi AR,2000.Heteroptera of Economic Importance.CRC Press,Florida.856 pp.
Son CK,Hwang SG,Hyun Y,Choi BS,2000.Field occurrence of stink bug and its damage in soybean.Korean J.Crop Sci.,45:405-410.
Sun HZ,Song YQ,Du J,Wang XD,Cheng ZJ,2016.Identification and tissue distribution of chemosensory protein and odorant binding protein genes in Athetis dissimilis(Lepidoptera:Noctuidae).Appl.Entomol.Zool.,51(3):409-420.
Tomokuni M,Yasunaga T,Takai M,Yamashita I,Kawamura M,Kawasawa T,1993.A Field Guide to Japanese Bugs:Terrestrial Heteropterans.Zenkoku Noson Kyoiku Kyokai,Tokyo.380 pp.
Vogt RG,Riddiford LM,1981.Pheromone binding and inactivation by moth antennae.Nature,293:161-163.
Vogt RG,Riddiford LM,Prestwich GD,1985.Kinetic properties of a sex pheromone-degrading enzyme:the sensillar esterase of Antheraea polyphemus.Proc.Natl.Acad.Sci.USA,82:8827-8831.
Vosshall LB,Stocker RF,2007.Molecular architecture of smell and taste in Drosophila.Annu.Rev.Neurosci.,30:505-533.
Xiao Y,Sun L,Ma XY,Dong K,Liu HW,Wang Q,Guo YY,Liu ZW,Zhang YJ,2016.Identification and characterization of the distinct expression profiles of candidate chemosensory membrane proteins in the antennal transcriptome of Adelphocoris lineolatus(Goeze).Insect Mol.Biol.,26(1):74-91.
Yuan HB,Ding YX,Gu SH,Sun L,Zhu XQ,Liu HW,Dhiloo KH,Zhang YJ,Guo YY,2015,Molecular characterization and expression profiling of odorant-binding proteins in Apolygus lucorum.PLo S ONE,10(10):e0140562.
Zhang QL,Yuan ML,2013.Progress in insect transcriptomics based on the next-generation sequencing technique.Acta Entomol.Sin.,56(12):1489-1508.[张棋麟,袁明龙,2013.基于新一代测序技术的昆虫转录组学研究进展.昆虫学报,56(12):1489-1508]
Zhang YN,Zhu XY,Zhang Q,Yin CY,Dong ZP,Zuo LH,Deng DG,Sun L,Li XM,2016.De novo assembly and characterization of antennal transcriptome reveal chemosensory system in Nysius ericae.J.Asia-Pac.Entomol.,19:1077-1087.
Zhou SH,Zhang J,Zhang SG,Zhang L,2008.Expression of chemosensory proteins in hairs on wings of Locusta migratoria(Orthoptera:Acrididae).J.Appl.Entomol.,132:439-450.