基于细胞色素C氧化酶亚基Ⅰ基因的小蜡螟系统进化分析
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摘要
【目的】小蜡螟Achroia Grisella Fabricius作为蜂业生产中的主要虫害,为确认它在鳞翅目昆虫中的遗传地位,本研究克隆了小蜡螟细胞色素C氧化酶亚基Ⅰ基因COⅠ,并对其序列进行相关生物信息学分析。【方法】根据NCBI已登录鳞翅目昆虫线粒体基因组mtDNA中COⅠ基因保守序列设计简并引物,以小蜡螟基因组为模板进行PCR,克隆并测序得到小蜡螟mtDNA CO Ⅰ基因序列(GenBank登录号:MF509586)。【结果】对CO Ⅰ基因编码区进行序列分析发现,其翻译的起始码子为TTG,以碱基T作为不完全终止;通过CO Ⅰ基因同源性和遗传距离分析发现,小蜡螟与大蜡螟Galleria mellonella同源性最高且遗传距离最近,分别为88.3%、0.117。系统进化分析发现,小蜡螟与其它鳞翅目昆虫聚为一支,其中螟蛾科昆虫大蜡螟、小蜡螟、米蛾Corcyra cephalonica和梢斑螟Corcyra cephalonica聚在一个小的分支。【结论】小蜡螟与大蜡螟的进化地位最为相近,也说明了利用CO Ⅰ基因可以区分同属不同种的蜡螟,研究结果为了解小蜡螟的遗传背景提供理论基础。
[Objectives] Achroia grisella Fabricius(small wax moth) is the main pests in the apiculture industry, in order to confirm its position of the genetic in Lepidoptera insects, cytochrome C oxidase subunit I gene(CO Ⅰ) of small wax moth was cloned, and analyzed by bioinformatics analysis. [Methods] According to the conserved sequence of mitochondrial genome mtDNA CO Ⅰ gene was logged in lepidoptera insects in NCBI, the degenerate primers were designed for the study, the genome of small wax moth as a template for PCR, mtDNA CO Ⅰ sequence(GenBank accession NO. MF509586) of small wax moth was cloned and sequenced. [Results] The sequence analysis of the CO Ⅰ gene coding region found that the initial code of its translation was TTG, and the base T was not terminated completely. The homology and genetic distance analysis of CO Ⅰ genes showed that Galleria mellonella(greater wax moth) and small wax moth were the most homologous and the genetic distance was 88.3% and 0.117 respectively. Phylogenetic analysis found that the small wax moth and other lepidoptera insects formed a separate clade, this clade gathered in a small branch that include greater wax moth, small wax moth, the rice moth(Corcyra cephalonica) and Corcyra cephalonica. [Conclusion] A. grisella was most closely related to G. mellonella, CO Ⅰ gene was used for distinguishing the different kinds of wax moths from same genus, the research of results provided a theoretical basis for understanding the genetic background of small wax moth.
[Objectives] Achroia grisella Fabricius(small wax moth) is the main pests in the apiculture industry, in order to confirm its position of the genetic in Lepidoptera insects, cytochrome C oxidase subunit I gene(CO Ⅰ) of small wax moth was cloned, and analyzed by bioinformatics analysis. [Methods] According to the conserved sequence of mitochondrial genome mtDNA CO Ⅰ gene was logged in lepidoptera insects in NCBI, the degenerate primers were designed for the study, the genome of small wax moth as a template for PCR, mtDNA CO Ⅰ sequence(GenBank accession NO. MF509586) of small wax moth was cloned and sequenced. [Results] The sequence analysis of the CO Ⅰ gene coding region found that the initial code of its translation was TTG, and the base T was not terminated completely. The homology and genetic distance analysis of CO Ⅰ genes showed that Galleria mellonella(greater wax moth) and small wax moth were the most homologous and the genetic distance was 88.3% and 0.117 respectively. Phylogenetic analysis found that the small wax moth and other lepidoptera insects formed a separate clade, this clade gathered in a small branch that include greater wax moth, small wax moth, the rice moth(Corcyra cephalonica) and Corcyra cephalonica. [Conclusion] A. grisella was most closely related to G. mellonella, CO Ⅰ gene was used for distinguishing the different kinds of wax moths from same genus, the research of results provided a theoretical basis for understanding the genetic background of small wax moth.
引文
Boore JL,1999.Animal mitochondrial genomes.Nucleic Acids Research,27(8):1767-1780.
Castro LR,Ruberu K,Dowton M,2006.Mitochondrial genomes of Vanhornia eucnemidarum(Apocrita:Vanhorniidae)and Primeuchroeus spp.(Aculeata:Chrysididae):Evidence of rearranged mitochondrial genomes within the Apocrita(Insecta:Hymenoptera).Genome,49(7):752-766.
Clary DO,Wahleithner JA,Wolstenholme DR,1983.Transfer RNA genes in Drosophila mitochondrial DNA:related 5'flanking sequences and comparisons to mammalian mitochondrial tRNA genes.Nucleic Acids Research,11(8):2411-2425.
Coates BS,Sumerford DV,Hellmich RL,2004.Geographic and voltinism differentiation among North American Ostrinia nubilalis(European corn borer)mitochondrial cytochrome c oxidase haplotypes.Journal of Insect Science,4(1):35-43.
Collet T,Ferreira KM,Arias MC,Soares AEE,Lama MAD,2006.Genetic structure of Africanized honeybee populations(Apis mellifera L.)from Brazil and Uruguay viewed through mitochondrial DNA CO I-CO II patterns.Heredity,97(5):329-335.
Crozier RH,Crozier YC,1993.The Mitochondrial Genome of the Honeybee Apis mellifera:Complete Sequence and Genome Organization.Genetics,133(1):97-117.
Feng F,1998.Biology and control of Galleria.Journal of Bee,1998(8):20-20.[冯峰,1998.蜡螟的生物学及防治.蜜蜂杂志,1998(8):20-20.]
Hoeh WR,Stewart DT,Sutherland BW,Zouros E,1996.Cytochrome c oxidase sequence comparisons suggest an unusually high rate of mitochondrial DNA evolution in Mytilus(Mollusca:Bivalvia).Molecular Biology&Evolution,13(2):418-421.
Lu C,Liu YQ,Liao SY,Li B,Xiang ZH,Han H,Wang XG,2002.Complete sequence determination and analysis of Bombyx mori mitochondrial genome.Journal of Agricultural Biotechnology,10(2):163-170.[鲁成,刘运强,廖顺尧,李斌,向仲怀,韩华,王学刚,2002.家蚕线粒体基因组全序列测定与分析.农业生物技术学报,10(2):163-170.]
Ojala D,Montoya J,Attardi G,1981.tRNA punctuation model of RNA processing in human mitochondria.Nature,290(5806):470-474.
Tamura K,Peterson D,Peterson N,Stecher G,Nei M,Kumar S,2011.MEGA5:molecular evolutionary genetics analysis using maximum likelihood,evolutionary distance,and maximum parsimony methods.Molecular Biology and Evolution,28(10):2731-2739.
Tan HW,Liu GH,Dong X,Lin RQ,Song HQ,Huang SY,Yuan ZG,Zhao GH,Zhu XQ,2011.The complete mitochondrial genome of the Asiatic cavity-nesting honeybee Apis cerana(Hymenoptera:Apidae).PLoS ONE,6(8):e23008.
Wang DM,Lin Y,2010.Application of mitochondrial DNA gene order in molecular systematics of insects.Guangdong Agricultural Sciences,37(6):188-190.[王德明,林杨,2010.线粒体DNA基因序列在昆虫分子系统学研究中的应用.广东农业科学,37(6):188-190.]
Wen B,2008.Achroia Grisella Fabricius and Galleria mellonella Linne.Apiculture of China,59(7):29-29.[文彬,2008.大蜡螟与小蜡螟.中国蜂业,59(7):29-29.]
Wolstenholme DR,1992.Animal mitochondrial DNA:structure and evolution.International Review of Cytology,141(6):173-216.
Wu YP,Li J,Zhao JL,Su TJ,Luo AR,Fan RJ,Chen MC,Wu CS,Zhu CD,2012.The complete mitochondrial genome of the rice moth,Corcyra cephalonica.Journal of Insect Science,12(72):1-14.
Xu ZY,1981.Determination of Achroia Grisella Fabricius and Galleria mellonella Linne.Apiculture of China,1981(6):7-8.[徐祖荫,1981.大小蜡螟的鉴别.中国蜂业,1981(6):7-8.]
Yin J,Hong GY,Wang AM,Cao YZ,Wei ZJ,2010.Mitochondrial genome of the cotton bollworm Helicoverpa armigera(Lepidoptera:Noctuidae)and comparison with other Lepidopterans.Mitochondrial DNA,21(5):160-169.
Castro LR,Ruberu K,Dowton M,2006.Mitochondrial genomes of Vanhornia eucnemidarum(Apocrita:Vanhorniidae)and Primeuchroeus spp.(Aculeata:Chrysididae):Evidence of rearranged mitochondrial genomes within the Apocrita(Insecta:Hymenoptera).Genome,49(7):752-766.
Clary DO,Wahleithner JA,Wolstenholme DR,1983.Transfer RNA genes in Drosophila mitochondrial DNA:related 5'flanking sequences and comparisons to mammalian mitochondrial tRNA genes.Nucleic Acids Research,11(8):2411-2425.
Coates BS,Sumerford DV,Hellmich RL,2004.Geographic and voltinism differentiation among North American Ostrinia nubilalis(European corn borer)mitochondrial cytochrome c oxidase haplotypes.Journal of Insect Science,4(1):35-43.
Collet T,Ferreira KM,Arias MC,Soares AEE,Lama MAD,2006.Genetic structure of Africanized honeybee populations(Apis mellifera L.)from Brazil and Uruguay viewed through mitochondrial DNA CO I-CO II patterns.Heredity,97(5):329-335.
Crozier RH,Crozier YC,1993.The Mitochondrial Genome of the Honeybee Apis mellifera:Complete Sequence and Genome Organization.Genetics,133(1):97-117.
Feng F,1998.Biology and control of Galleria.Journal of Bee,1998(8):20-20.[冯峰,1998.蜡螟的生物学及防治.蜜蜂杂志,1998(8):20-20.]
Hoeh WR,Stewart DT,Sutherland BW,Zouros E,1996.Cytochrome c oxidase sequence comparisons suggest an unusually high rate of mitochondrial DNA evolution in Mytilus(Mollusca:Bivalvia).Molecular Biology&Evolution,13(2):418-421.
Lu C,Liu YQ,Liao SY,Li B,Xiang ZH,Han H,Wang XG,2002.Complete sequence determination and analysis of Bombyx mori mitochondrial genome.Journal of Agricultural Biotechnology,10(2):163-170.[鲁成,刘运强,廖顺尧,李斌,向仲怀,韩华,王学刚,2002.家蚕线粒体基因组全序列测定与分析.农业生物技术学报,10(2):163-170.]
Ojala D,Montoya J,Attardi G,1981.tRNA punctuation model of RNA processing in human mitochondria.Nature,290(5806):470-474.
Tamura K,Peterson D,Peterson N,Stecher G,Nei M,Kumar S,2011.MEGA5:molecular evolutionary genetics analysis using maximum likelihood,evolutionary distance,and maximum parsimony methods.Molecular Biology and Evolution,28(10):2731-2739.
Tan HW,Liu GH,Dong X,Lin RQ,Song HQ,Huang SY,Yuan ZG,Zhao GH,Zhu XQ,2011.The complete mitochondrial genome of the Asiatic cavity-nesting honeybee Apis cerana(Hymenoptera:Apidae).PLoS ONE,6(8):e23008.
Wang DM,Lin Y,2010.Application of mitochondrial DNA gene order in molecular systematics of insects.Guangdong Agricultural Sciences,37(6):188-190.[王德明,林杨,2010.线粒体DNA基因序列在昆虫分子系统学研究中的应用.广东农业科学,37(6):188-190.]
Wen B,2008.Achroia Grisella Fabricius and Galleria mellonella Linne.Apiculture of China,59(7):29-29.[文彬,2008.大蜡螟与小蜡螟.中国蜂业,59(7):29-29.]
Wolstenholme DR,1992.Animal mitochondrial DNA:structure and evolution.International Review of Cytology,141(6):173-216.
Wu YP,Li J,Zhao JL,Su TJ,Luo AR,Fan RJ,Chen MC,Wu CS,Zhu CD,2012.The complete mitochondrial genome of the rice moth,Corcyra cephalonica.Journal of Insect Science,12(72):1-14.
Xu ZY,1981.Determination of Achroia Grisella Fabricius and Galleria mellonella Linne.Apiculture of China,1981(6):7-8.[徐祖荫,1981.大小蜡螟的鉴别.中国蜂业,1981(6):7-8.]
Yin J,Hong GY,Wang AM,Cao YZ,Wei ZJ,2010.Mitochondrial genome of the cotton bollworm Helicoverpa armigera(Lepidoptera:Noctuidae)and comparison with other Lepidopterans.Mitochondrial DNA,21(5):160-169.