基于28S rDNA分析板栗和杉木上针叶小爪螨物种分化原因
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摘要
【目的】我国经济林树种板栗和杉木上的重要害螨一直被认为是同一种螨——针叶小爪螨,但针叶小爪螨的山东板栗种群和浙江杉木种群间存在生殖隔离,且生殖隔离并非由地理隔离和能调控寄主生殖的内共生菌引起,似乎二者已分化为2个独立的种。本研究目的是明确其成种原因,丰富物种形成理论。【方法】从之前做杂交试验时所采集的针叶小爪螨板栗和杉木种群中各随机取3头雌成螨,提取单头雌成螨的总DNA,使用根据其他小爪螨28S rDNA两端保守序列设计的引物扩增28S rDNA,扩增产物纯化后测序。比较2种群的28S rDNA序列,并基于28S rDNA序列构建小爪螨属系统发育树,结合已有研究分析两种群分化原因。【结果】每个种群的3个个体的28S rDNA序列完全一致,无种群内变异。2种群的28S rDNA序列一致性为98. 3%,遗传距离为1. 7%。在基于28S rDNA序列构建的小爪螨属系统发育树上,浙江杉木种群与采自日本的日本柳杉上的本岛小爪螨亲缘关系最近,山东板栗种群与采自日本的日本栗上的栗小爪螨的亲缘关系最近。【结论】板栗和杉木上的叶螨并非由同一种叶螨因适应不同寄主植物分化而成,很可能是2个独立的物种。
【Objective】 The important pest mites that infest Castanea mollissima,an economic forest species,and Cunninghamia lanceolata,have been considered to be the same mites,Oligonychus ununguis. However,previous studies have shown that reproductive isolation exists between the C. mollissima population from Shandong Province(TSBL) and the C. lanceolata population from Zhejiang Province(ZJSM) of O. ununguis and the reproductive isolation is not caused by geographic isolation and the endosymbionts that can manipulate the reproduction of their hosts. The two populations have been differentiated into two independent species,but whether the differentiation is related to adaptation to different host plants has not been determined. This study was carried out to clarify the differentiation reasons and enrich the speciation theory.【Method】Three female adult mites were randomly selected from the C. mollissima population and the C. lanceolata population respectively which collected previously to be used to hybridization experiment. The genomic DNA was extracted from a single female adult. The 28S rDNA sequences were amplified used the primers designed according to the conserved sequences of the 28S rDNA of other mites in Oligonychus. The amplified products were purified and sequenced. The 28S rDNA sequences of the two populations were compared and the phylogenetic tree of Oligonychus was constructed based on the 28S rDNA sequences. The differentiation reasons of these two populations were analyzed combined with previous studies.【Result】 The 28S rDNA sequences of three individuals in each population were identical,without intraspecific variation. The interpopulation sequence identity and the genetic distance between the C. mollissima population and the C.lanceolata population based on 28S rDNA sequences were 98. 3% and 1. 7% respectively. The phylogenetic tree of Oligonychus based on 28S rDNA sequences showed that O. hondoensis collected from Cryptomeria japonica in Japan was the closest relative of the C. lanceolata population,while O. castaneae collected from Castanea crenata in Japan was the closest relative of the C. mollissima population.【Conclusion】 The spider mites on C. mollissima and C. lanceolata are probably two independent species rather than differentiated from the same kind of spider mite because of adaptation to different host plants.
【Objective】 The important pest mites that infest Castanea mollissima,an economic forest species,and Cunninghamia lanceolata,have been considered to be the same mites,Oligonychus ununguis. However,previous studies have shown that reproductive isolation exists between the C. mollissima population from Shandong Province(TSBL) and the C. lanceolata population from Zhejiang Province(ZJSM) of O. ununguis and the reproductive isolation is not caused by geographic isolation and the endosymbionts that can manipulate the reproduction of their hosts. The two populations have been differentiated into two independent species,but whether the differentiation is related to adaptation to different host plants has not been determined. This study was carried out to clarify the differentiation reasons and enrich the speciation theory.【Method】Three female adult mites were randomly selected from the C. mollissima population and the C. lanceolata population respectively which collected previously to be used to hybridization experiment. The genomic DNA was extracted from a single female adult. The 28S rDNA sequences were amplified used the primers designed according to the conserved sequences of the 28S rDNA of other mites in Oligonychus. The amplified products were purified and sequenced. The 28S rDNA sequences of the two populations were compared and the phylogenetic tree of Oligonychus was constructed based on the 28S rDNA sequences. The differentiation reasons of these two populations were analyzed combined with previous studies.【Result】 The 28S rDNA sequences of three individuals in each population were identical,without intraspecific variation. The interpopulation sequence identity and the genetic distance between the C. mollissima population and the C.lanceolata population based on 28S rDNA sequences were 98. 3% and 1. 7% respectively. The phylogenetic tree of Oligonychus based on 28S rDNA sequences showed that O. hondoensis collected from Cryptomeria japonica in Japan was the closest relative of the C. lanceolata population,while O. castaneae collected from Castanea crenata in Japan was the closest relative of the C. mollissima population.【Conclusion】 The spider mites on C. mollissima and C. lanceolata are probably two independent species rather than differentiated from the same kind of spider mite because of adaptation to different host plants.
引文
江世宏,陈晓琴,吴深健,等.2009.基于28S r DNA的叩甲科分子系统发育关系研究.昆虫学报,52(1):74-83.(Jiang S H,Chen X Q,Wu S J,et al.2009.Molecular phylogenetic analysis of Elateridae(Insecta:Coleoptera)based on 28S r DNAgene fragments.Acta Entomologica Sinica,52(1):74-83.[in Chinese])
李国庆,于明志,洪晓月.2010.基于核糖体28S rRNA对叶螨的鉴定及其系统发育分析.南京农业大学学报,33(5):49-54.(Li G Q,Yu M Z,Hong X Y.2010.Identification and phylogenetic analysis of spider mites based on 28S rRNA sequences.Journal of Nanjing Agricultural University,33(5):49-54.[in Chinese])
刘颖,谢蓉蓉,洪晓月.2010.共生菌Cardinium对朱砂叶螨的生殖调控作用.昆虫学报,53(11):1233-1240.(Liu Y,Xie R R,Hong X Y.2010.Manipulation of symbiont Cardinium on the reproduction of the carmine spider mite,Tetranychus cinnabarinus(Boisduval)(Acari:Tetranychidae).Acta Entomologica Sinica,53(11):1233-1240.[in Chinese])
陆明红,谢蓉蓉,赵臻君,等.2011.二斑叶螨两种群中Wolbachia诱导的胞质不亲和作用的影响因子比较研究.昆虫学报,54(9):1018-1026.(Lu M H,Xie R R,Zhao Z J,et al.2011.A comparative study of factors influencing the expression of Wolbachia-induced cytoplasmic incompatibility in two populations of the two-spotted spider mite,Tetranychus urticae Koch(Acari:Tetranychidae).Acta Entomologica Sinica,54(9):1018-1026.[in Chinese])
任保青,陈之端.2010.植物DNA条形码技术.植物学报,5(1):1-12.(Ren B Q,Chen Z D.2010.DNA barcoding plant life.Chinese Bulletin of Botany,45(1):1-12.[in Chinese])
孙菲,杨春红,李波,等.2017.针叶小爪螨不同种群内共生菌Wolbachia和Cardinium的检测及系统发育分析.林业科学,53(12):12-19.(Sun F,Yang C H,Li B,et al.2017.Detection of Wolbachia and Cardinium endosymbionts in different Oligonychus ununguis populations and phylogenetic analysis of Cardinium.Scientia Silvae Sinicae,53(12):12-19.[in Chinese])
孙绪艮,苗良.1990.针叶小爪螨研究初报.山东农业大学学报,21(3):41-46.(Sun X G,Miao L.1990.Studies on Oligonychus ununguis(Jacobi).Journal of Shandong Agricultural University,21(3):41-46.[in Chinese])
孙绪艮,徐常青,周成刚,等.2000.针叶小爪螨不同种群在针叶树和阔叶树上的生长发育和繁殖及其生殖隔离.昆虫学报,43(1):52-57.(Sun X G,Xu C Q,Zhou C G,et al.2000.Performances and reproductive isolation of different populations of Oligonychus ununguis(Jacobi)on conifer and broadleaf trees.Acta Entomologica Sinica,43(1):52-57.[in Chinese])
王慧芙.1981.中国经济昆虫志.第二十三册.螨目叶螨总科.北京:科学出版社.(Wang H F.1981.Economic insect fauna of China,Fasc.23,Acariformes:Tetranychoidea.Beijing:Science Press.[in Chinese])
尹淑艳,徐常青,崔孝平,等.2010a.针叶小爪螨板栗和杉木种群的形态差异.林业科学,46(6):166-170.(Yin S Y,Xu C Q,Cui X P,et al.2010a.Morphological differences of Oligonychus ununguis populations from Castanea mollissima and Cunninghamia lanceolata.Scientia Silvae Sinicae,46(6):166-170.[in Chinese])
尹淑艳,于新社,郭慧玲,等.2010b.针叶小爪螨板栗和杉木种群间的遗传分化和杂交试验.昆虫学报,53(5):555-563.(Yin S Y,Yu X S,Guo H L,et al.2010b.Genetic differentiation and cross-breeding experiments between Oligonychus ununguis(Acari:Tetranychidae)populations from host plants Castanea mollissima and Cunninghamia lanceolata.Acta Entomologica Sinica,53(5):555-563.[in Chinese])
张时敏.1982.针叶小爪螨初步观察.林业科技通讯,(3):30-32.(Zhang S M.1982.Preliminary observation on Oligonychus ununguis.Forest Science and Technology,(3):30-32.[in Chinese])
张艳凯,孙兵,洪晓月.2014.Wolbachia在山楂双叶螨中的感染及对寄主生殖的影响.昆虫学报,57(8):914-920.(Zhang Y K,Sun B,Hong X Y.2014.Infection and reproductive effects of Wolbachia in the hawthorn spider mite,Amphitetranychus viennensis(Acarina:Tetranychidae).Acta Entomologica Sinica,57(8):914-920.[in Chinese])
Bolland H R,Gutierrez J,Flechtmann C H W.1998.World catalogue of the spider mites family Acari:Tetranychidae with references to taxonomy,synonymy,host pests and distribution.Leiden:Academic Publ Brill.
Ehara S,Gotoh T.2011.Two new species of spider mites occuring in Japan(Acari,Tetranychidae).J Acarol Soc Jpn,5(1):17-25.
Gotoh T,Noda H,Ito S.2007.Cardinium symbionts cause cytoplasmic incompatibility in spider mites.Heredity,98(1):13-20.
Groot T V M,Breeuwer J A J.2006.Cardinium symbionts induce haploid thelytoky in most clones of three closely related Brevipalpus species.Exp Appl Acarol,39(3/4):257-271.
Hebert P D N,Cywinska A,Ball S L,et al.2003.Biological identifications through DNA barcodes.Proc R Soc B,270(1512):313-321.
Kanouh M,Tixier M S,Guichou S,et al.2010.Two synonymy cases within the genus Neoseiulella.Biological Journal of the Linnean Society,101(2):323-344.
Lehmitz R,Decker P.2017.The nuclear 28S gene fragment D3 as species marker in oribatid mites(Acari,Oribatida)from German peatlands.Experimental&Applied Acarology,71(3):259-276.
Matsuda T,Hinomoto N,Singh R N,et al.2012.Molecular-based identification and phylogeny of Oligonychus species(Acari:Tetranychidae).Journal of Economic Entomology,105(3):1043-1050.
Matsuda T,Morishita M,Hinomoto N,et al.2014.Phylogenetic Analysis of the spider mite sub-Family Tetranychinae(Acari:Tetranychidae)based on the mitochondrial COI gene and the 18Sand the 5'end of the 28S rRNA genes indicates that several genera are polyphyletic.PLoS ONE,9(10):e108672.
Migeon A,Dorkeld F.[2017-08-25].Spider Mites Web:a comprehensive database for the Tetranychidae.http://www.montpellier.inra.Fr/CBGP/spmweb.
Moritz C,Cicero C.2004.DNA barcoding:promise and pitfalls.PLoSBiol,2(10):e354.
O’Neill S L,Giordano R,Colbert A M,et al.1992.16S rRNAphylogenetic analysis of the bacterial endosymbionts associated with cytoplasmic incompatibility in insects.Proc Natl Acad Sci USA,89(7):2699-702.
Ros V I D,Breeuwer J A J.2007.Spider mite(Acari:Tetranychidae)mitochondrial COI phylogeny reviewed:host plant relationships,phylogeography,reproductive parasites and barcoding.Exp Appl Acarol,42(4):239-262.
Skoracka A,Dabert M.2010.The cereal rust mite Abacarus hystrix(Acari:Eriophyoidea)is a complex of species evidence from mitochondrial and nuclear DNA sequences.Bulletin of Entomological Research,100(3):263-272.
Vargas M,Garcia-Varela M,Laclette J P,et al.2005.Application of ITS-2 sequences as markers for identification and phylogenetic inference within the genus Geomylichus(Acari:Listrophoridae).Exp Appl Acarol,35(3):223-238.
Via S,1990.Ecological genetics and host adaptation in herbivorous insects:the experimental study of evolution in natural and agricultural systems.Annual Review of Entomology,35(35):421-446.
Werren J H,Baldo L,Clark M E.2008.Wolbachia:master manipulators of invertebrate biology.Nat Rev Microbiol,6(10):741-751.
Xie R R,Liu Y,Hong X Y,et al.2006.Effect of infection rate of Wolbachia on the reproduction in Tetranychus kanzawai Kishida(Acari:Tetranychidae)in China.Int J Acarol,32(4):407-415.
Xie R R,Zhou L L,Zhao Z J,et al.2010.Male age influences the strength of Cardinium-induced cytoplasmic incompatibility expression in the carmine spider mite Tetranychus cinnabarinus.Appl Entomol Zool,45(3):417-423.
Zhu L Y,Zhang K J,Zhang Y K,et al.2012.Wolbachia strengthens cardinium-induced cytoplasmic incompatibility in the spider mite Tetranychus piercei Mc Gregor.Curr Microbiol,65(5):516-523.
李国庆,于明志,洪晓月.2010.基于核糖体28S rRNA对叶螨的鉴定及其系统发育分析.南京农业大学学报,33(5):49-54.(Li G Q,Yu M Z,Hong X Y.2010.Identification and phylogenetic analysis of spider mites based on 28S rRNA sequences.Journal of Nanjing Agricultural University,33(5):49-54.[in Chinese])
刘颖,谢蓉蓉,洪晓月.2010.共生菌Cardinium对朱砂叶螨的生殖调控作用.昆虫学报,53(11):1233-1240.(Liu Y,Xie R R,Hong X Y.2010.Manipulation of symbiont Cardinium on the reproduction of the carmine spider mite,Tetranychus cinnabarinus(Boisduval)(Acari:Tetranychidae).Acta Entomologica Sinica,53(11):1233-1240.[in Chinese])
陆明红,谢蓉蓉,赵臻君,等.2011.二斑叶螨两种群中Wolbachia诱导的胞质不亲和作用的影响因子比较研究.昆虫学报,54(9):1018-1026.(Lu M H,Xie R R,Zhao Z J,et al.2011.A comparative study of factors influencing the expression of Wolbachia-induced cytoplasmic incompatibility in two populations of the two-spotted spider mite,Tetranychus urticae Koch(Acari:Tetranychidae).Acta Entomologica Sinica,54(9):1018-1026.[in Chinese])
任保青,陈之端.2010.植物DNA条形码技术.植物学报,5(1):1-12.(Ren B Q,Chen Z D.2010.DNA barcoding plant life.Chinese Bulletin of Botany,45(1):1-12.[in Chinese])
孙菲,杨春红,李波,等.2017.针叶小爪螨不同种群内共生菌Wolbachia和Cardinium的检测及系统发育分析.林业科学,53(12):12-19.(Sun F,Yang C H,Li B,et al.2017.Detection of Wolbachia and Cardinium endosymbionts in different Oligonychus ununguis populations and phylogenetic analysis of Cardinium.Scientia Silvae Sinicae,53(12):12-19.[in Chinese])
孙绪艮,苗良.1990.针叶小爪螨研究初报.山东农业大学学报,21(3):41-46.(Sun X G,Miao L.1990.Studies on Oligonychus ununguis(Jacobi).Journal of Shandong Agricultural University,21(3):41-46.[in Chinese])
孙绪艮,徐常青,周成刚,等.2000.针叶小爪螨不同种群在针叶树和阔叶树上的生长发育和繁殖及其生殖隔离.昆虫学报,43(1):52-57.(Sun X G,Xu C Q,Zhou C G,et al.2000.Performances and reproductive isolation of different populations of Oligonychus ununguis(Jacobi)on conifer and broadleaf trees.Acta Entomologica Sinica,43(1):52-57.[in Chinese])
王慧芙.1981.中国经济昆虫志.第二十三册.螨目叶螨总科.北京:科学出版社.(Wang H F.1981.Economic insect fauna of China,Fasc.23,Acariformes:Tetranychoidea.Beijing:Science Press.[in Chinese])
尹淑艳,徐常青,崔孝平,等.2010a.针叶小爪螨板栗和杉木种群的形态差异.林业科学,46(6):166-170.(Yin S Y,Xu C Q,Cui X P,et al.2010a.Morphological differences of Oligonychus ununguis populations from Castanea mollissima and Cunninghamia lanceolata.Scientia Silvae Sinicae,46(6):166-170.[in Chinese])
尹淑艳,于新社,郭慧玲,等.2010b.针叶小爪螨板栗和杉木种群间的遗传分化和杂交试验.昆虫学报,53(5):555-563.(Yin S Y,Yu X S,Guo H L,et al.2010b.Genetic differentiation and cross-breeding experiments between Oligonychus ununguis(Acari:Tetranychidae)populations from host plants Castanea mollissima and Cunninghamia lanceolata.Acta Entomologica Sinica,53(5):555-563.[in Chinese])
张时敏.1982.针叶小爪螨初步观察.林业科技通讯,(3):30-32.(Zhang S M.1982.Preliminary observation on Oligonychus ununguis.Forest Science and Technology,(3):30-32.[in Chinese])
张艳凯,孙兵,洪晓月.2014.Wolbachia在山楂双叶螨中的感染及对寄主生殖的影响.昆虫学报,57(8):914-920.(Zhang Y K,Sun B,Hong X Y.2014.Infection and reproductive effects of Wolbachia in the hawthorn spider mite,Amphitetranychus viennensis(Acarina:Tetranychidae).Acta Entomologica Sinica,57(8):914-920.[in Chinese])
Bolland H R,Gutierrez J,Flechtmann C H W.1998.World catalogue of the spider mites family Acari:Tetranychidae with references to taxonomy,synonymy,host pests and distribution.Leiden:Academic Publ Brill.
Ehara S,Gotoh T.2011.Two new species of spider mites occuring in Japan(Acari,Tetranychidae).J Acarol Soc Jpn,5(1):17-25.
Gotoh T,Noda H,Ito S.2007.Cardinium symbionts cause cytoplasmic incompatibility in spider mites.Heredity,98(1):13-20.
Groot T V M,Breeuwer J A J.2006.Cardinium symbionts induce haploid thelytoky in most clones of three closely related Brevipalpus species.Exp Appl Acarol,39(3/4):257-271.
Hebert P D N,Cywinska A,Ball S L,et al.2003.Biological identifications through DNA barcodes.Proc R Soc B,270(1512):313-321.
Kanouh M,Tixier M S,Guichou S,et al.2010.Two synonymy cases within the genus Neoseiulella.Biological Journal of the Linnean Society,101(2):323-344.
Lehmitz R,Decker P.2017.The nuclear 28S gene fragment D3 as species marker in oribatid mites(Acari,Oribatida)from German peatlands.Experimental&Applied Acarology,71(3):259-276.
Matsuda T,Hinomoto N,Singh R N,et al.2012.Molecular-based identification and phylogeny of Oligonychus species(Acari:Tetranychidae).Journal of Economic Entomology,105(3):1043-1050.
Matsuda T,Morishita M,Hinomoto N,et al.2014.Phylogenetic Analysis of the spider mite sub-Family Tetranychinae(Acari:Tetranychidae)based on the mitochondrial COI gene and the 18Sand the 5'end of the 28S rRNA genes indicates that several genera are polyphyletic.PLoS ONE,9(10):e108672.
Migeon A,Dorkeld F.[2017-08-25].Spider Mites Web:a comprehensive database for the Tetranychidae.http://www.montpellier.inra.Fr/CBGP/spmweb.
Moritz C,Cicero C.2004.DNA barcoding:promise and pitfalls.PLoSBiol,2(10):e354.
O’Neill S L,Giordano R,Colbert A M,et al.1992.16S rRNAphylogenetic analysis of the bacterial endosymbionts associated with cytoplasmic incompatibility in insects.Proc Natl Acad Sci USA,89(7):2699-702.
Ros V I D,Breeuwer J A J.2007.Spider mite(Acari:Tetranychidae)mitochondrial COI phylogeny reviewed:host plant relationships,phylogeography,reproductive parasites and barcoding.Exp Appl Acarol,42(4):239-262.
Skoracka A,Dabert M.2010.The cereal rust mite Abacarus hystrix(Acari:Eriophyoidea)is a complex of species evidence from mitochondrial and nuclear DNA sequences.Bulletin of Entomological Research,100(3):263-272.
Vargas M,Garcia-Varela M,Laclette J P,et al.2005.Application of ITS-2 sequences as markers for identification and phylogenetic inference within the genus Geomylichus(Acari:Listrophoridae).Exp Appl Acarol,35(3):223-238.
Via S,1990.Ecological genetics and host adaptation in herbivorous insects:the experimental study of evolution in natural and agricultural systems.Annual Review of Entomology,35(35):421-446.
Werren J H,Baldo L,Clark M E.2008.Wolbachia:master manipulators of invertebrate biology.Nat Rev Microbiol,6(10):741-751.
Xie R R,Liu Y,Hong X Y,et al.2006.Effect of infection rate of Wolbachia on the reproduction in Tetranychus kanzawai Kishida(Acari:Tetranychidae)in China.Int J Acarol,32(4):407-415.
Xie R R,Zhou L L,Zhao Z J,et al.2010.Male age influences the strength of Cardinium-induced cytoplasmic incompatibility expression in the carmine spider mite Tetranychus cinnabarinus.Appl Entomol Zool,45(3):417-423.
Zhu L Y,Zhang K J,Zhang Y K,et al.2012.Wolbachia strengthens cardinium-induced cytoplasmic incompatibility in the spider mite Tetranychus piercei Mc Gregor.Curr Microbiol,65(5):516-523.