【目的】探明地方鸡种与引进鸡种遗传多态性特点,探讨COⅠ这一特定基因的特定区段作为DNA条形码在识别鸡种方面的可行性和有效性。【方法】以13个中国地方鸡种和2个国外引进品种为研究对象,利用DNA测序技术测定了线粒体细胞色素C氧化酶...【目的】探明地方鸡种与引进鸡种遗传多态性特点,探讨COⅠ这一特定基因的特定区段作为DNA条形码在识别鸡种方面的可行性和有效性。【方法】以13个中国地方鸡种和2个国外引进品种为研究对象,利用DNA测序技术测定了线粒体细胞色素C氧化酶亚基Ⅰ(cytochrome c oxidaseⅠ,COⅠ)基因部分序列。【结果】选择的这段COⅠ基因序列有38个突变位点,15个鸡种单倍型多样度平均为0.963,核苷酸多样度平均为0.00518,其中引进鸡种明显低于地方鸡种(除藏鸡外);15个鸡种品种间Kimura双参数遗传距离为0.056%—0.917%,种内遗传距离为0—0.346%,15个鸡种的DNA分类和形态学分类基本一致,引进鸡种与地方鸡种分歧较远。【结论】利用线粒体细胞色素C氧化酶Ⅰ(COⅠ)这一特定基因的特定区段来做DNA条形编码的基础,进行不同鸡品种鉴定,具有可行性和有效性。展开更多
利用mt DNA COⅠ基因片段(有效长度635 bp)对来自达里湖(DL)、岗更湖(GG)、呼伦湖(HL)、黑龙江(HLJ)、松花江(SHJ)和乌苏里江(WSL)6个地理群体共72个样品进行了遗传多样性和遗传结构分析。从碱基组成来看,6个地理群体在COⅠ区段上的A+T...利用mt DNA COⅠ基因片段(有效长度635 bp)对来自达里湖(DL)、岗更湖(GG)、呼伦湖(HL)、黑龙江(HLJ)、松花江(SHJ)和乌苏里江(WSL)6个地理群体共72个样品进行了遗传多样性和遗传结构分析。从碱基组成来看,6个地理群体在COⅠ区段上的A+T含量(52.37%)明显高于G+C含量(47.67%),符合脊椎动物的碱基组成特点;从各项遗传参数来看,达里湖流域2个群体(DL和GG)的遗传多样性低于黑龙江流域4个群体(HL、HLJ、SHJ、WSL);6个群体共检测到20个单倍型,并共享1个单倍型,达里湖流域群体共享1个单倍型,黑龙江流域群体共享1个单倍型。虽然达里湖流域和黑龙江流域各群体间遗传分化明显(P<0.05),遗传结构存在差异,但AMOVA检验不支持这种遗传差异;另外,群体间频繁的基因交流进一步证实,6个地理群体亲缘关系近,有共同的起源,尚未达到分化为不同的种或亚种的水平。展开更多
【目的】研究贵州省茶棍蓟马不同地理种群间的遗传多样性,为掌握贵州省茶棍蓟马的遗传动态和扩散规律及制定防控措施提供理论依据。【方法】以贵州省11个不同地理区域的茶棍蓟马种群mtDNA CO Ⅰ和mtDNA CO Ⅱ基因序列为靶标,利用MEGA 6....【目的】研究贵州省茶棍蓟马不同地理种群间的遗传多样性,为掌握贵州省茶棍蓟马的遗传动态和扩散规律及制定防控措施提供理论依据。【方法】以贵州省11个不同地理区域的茶棍蓟马种群mtDNA CO Ⅰ和mtDNA CO Ⅱ基因序列为靶标,利用MEGA 6.0、DnaSP 5.10、Arlequin 3.5.2.2和Network 2.0等软件对种群遗传分化、基因流水平、分子变异及种群遗传多样性等进行分析。【结果】基于mtDNA CO Ⅰ和mtDNA CO Ⅱ基因分析时,分别检测到11种和9种单倍型,各地理种群的单倍型多样度(Hd)较高,分别为0.609和0.633;总体遗传固定指数(FST)分别为0.11902和0.09052,基因流(Nm)分别为2.00和2.51,表明贵州省茶棍蓟马各地理种群间的基因交流水平较高,种群间遗传分化较小。mtDNA CO Ⅰ和mtDNA CO Ⅱ基因序列的单倍型网状树分析均无明显分支;种群间的分子变异(AMOVA)分析结果表明,茶棍蓟马的遗传变异主要来自种群内部。总群体的中性检验Fu’s Fs不显著(P>0.05),且错配分布曲线呈现多峰,说明贵州省茶棍蓟马种群在较近的历史时期内保持相对稳定,未经历明显的种群扩张,或处于临界状态。【结论】贵州省茶棍蓟马种群遗传多样性较丰富,虽然尚未形成明显的种群扩张,但可能已处于临界状态,应积极采取综合防控措施,力争将茶棍蓟马种群数量控制在危害水平以下,以保证贵州省茶产业生产健康可持续发展。展开更多
作者于2009年在青岛农业大学昆虫实验室内,利用引物C1-J-1718和2195扩增线粒体DNA的COⅠ基因并对所得到的片段进行序列分析,研究了苹果小卷叶蛾Adoxophyes orana Fischer von Roslerstamm胶东、北京、陕西3个地理种群的遗传分化,并比较...作者于2009年在青岛农业大学昆虫实验室内,利用引物C1-J-1718和2195扩增线粒体DNA的COⅠ基因并对所得到的片段进行序列分析,研究了苹果小卷叶蛾Adoxophyes orana Fischer von Roslerstamm胶东、北京、陕西3个地理种群的遗传分化,并比较了上述两个片段在种群水平系统发育研究中的应用价值。研究结果表明,C1-J-1718扩增序列更适合苹果小卷叶蛾的系统发育研究。胶东种群与北京种群的遗传距离为0.059,与陕西种群为0.049,北京种群与陕西种群为0.047,不同的地理种群遗传分化明显;胶东种群内青岛、威海、烟台地理种群间不存在遗传分化。研究结果还表明:北京种群与A.orana的遗传距离为0;陕西种群的与A.orana like的遗传距离仅为0.007。展开更多
为了揭示吉林山鸡细胞色素C氧化酶亚基Ⅰ(COⅠ)基因的遗传多样性及其与地方鸡种的遗传关系,试验采用DNA测序技术测定吉林山鸡线粒体COⅠ基因序列。结果表明:20只山鸡样本mt DNA COⅠ基因序列中,共检测到核苷酸多态位点19个,20个个体具有...为了揭示吉林山鸡细胞色素C氧化酶亚基Ⅰ(COⅠ)基因的遗传多样性及其与地方鸡种的遗传关系,试验采用DNA测序技术测定吉林山鸡线粒体COⅠ基因序列。结果表明:20只山鸡样本mt DNA COⅠ基因序列中,共检测到核苷酸多态位点19个,20个个体具有4种单倍型,吉林山鸡群体内平均核苷酸多样度(Pi)为0.002 88,单倍型多样性(Hd)为0.625,可以作为品种鉴定的依据。吉林山鸡种群的遗传多样性水平较低,建议对吉林山鸡种群的保护制订更有效的繁殖计划,避免近亲杂交。展开更多
To determine the role of mitochondrial COⅠgene in classification and identification of species,a total of 39 single individuals from nine species pertaining to two genera of Cynoglossidae in the Yangtze Estuary were ...To determine the role of mitochondrial COⅠgene in classification and identification of species,a total of 39 single individuals from nine species pertaining to two genera of Cynoglossidae in the Yangtze Estuary were barcoded by COⅠ,sequenced and compared with that of other Cynoglossidae species recorded in the Gen Bank. Total genomic DNA was extracted from each scale sample using the classic phenol / chloroform extraction method. Six hundred and fifty base pairs( bp)COⅠfragments were amplified using the primers ’i. e. ’ F1: 5’- TCA ACC AAC CAC AAA GAC ATT GGC AC- 3’,R1: 5’- TAG ACT TCT GGG TGG CCA AAG AAT CA- 3’. Every PCR amplification was performed in a total volume of 50 μL of PCR mixture. PCR products were purified and then sequenced in both forward and reverse directions using an ABI PRISMTM 3730 XL Automated Sequencer. DNA sequences were aligned with clustal W using default parameters. Base composition,variable and parsimony informative sites were determined using MEGA 5. 0. Neighbor- joining( NJ) and Maximum parsimony( MP) phylogenetic trees were constructed for COⅠhaplotypes( Kimura 2 Parameter substitution model,K2P; 1 000 bootstraps pseudoreplications) using MEGA 5. 0. Using the MEGA5. 0 software for statistical analysis,the averaged AT content was greater than the GC content( Tab. 2). The GC content of codon position 1 averaged 53. 8%( 51. 8%-57. 3%),which of position 2 for 42. 0%,and that of position 3 ranged from 28. 1% to 37. 8% in average of 32. 4%( Tab. 4). The transitional pairs( si) was slightly more than the transversional pairs( sv),and the ratio( R = si/sv) was 1. 45( Tab. 3). Analysis of the frequency of amino acids in COⅠgene encoding protein showed that the highest frequency of amino acid was leucine,and the lowest frequency of amino acid was tryptophan( Tab. 5). The average K2 P distances pairwise-species and within-species were 0. 191 and 0. 003,respectively( Tab. 6). The K2 P distance pairwise-species was 63. 7 times of that within-species. According to the MP and NJ trees for all 39 sequences,it demonstrated that Cynoglossinae in the Yangtze Estuary was a monophyletic group( Fig. 1). However,the phylogenetic relationships revealed by the COⅠsequence analyses were not consistent with those inferred from morphological classification. In contrast to morphological classification,Paraplagusis japonica of Rhinoplagusia and Cynoglossus robustus of Cynoglossoides were placed in a same clade with support of a high bootstrap value( Fig. 1). The species in the sub-genera of Areliscus clustered on an independent branch. But synonymic phenomena existed in the following two groups of species: C. abbreviatus and C. purpureomaculatus,C. lighti and C. joyner,with the pairwise-species of 0. 002 and 0. 007,respectively( Tab. 6). Our results highlight that the information from COⅠsequences not only can filter out the synonym of the same species,but also be able to be used to carry out effective identification for Cynoglossidae species,which further shows that mitochondrial COⅠis feasible as the classification barcode.展开更多
运用一对特异性引物扩增并测定了南海硬骨鱼类40个物种89个样本的线粒体DNA的细胞色素c氧化酶Ⅰ(COⅠ)约660bp的部分序列,即COⅠ条码序列。综合比较了《中国鱼类系统检索》、《海洋鱼类志》、FishBase鱼类形态学分类库、ITIS综合分类学...运用一对特异性引物扩增并测定了南海硬骨鱼类40个物种89个样本的线粒体DNA的细胞色素c氧化酶Ⅰ(COⅠ)约660bp的部分序列,即COⅠ条码序列。综合比较了《中国鱼类系统检索》、《海洋鱼类志》、FishBase鱼类形态学分类库、ITIS综合分类学信息系统和生物条码系统(The Barcode of Life Data System,BOLD)的相应形态学与DNA序列资料,在揭示完善我国鱼类检索系统必要性基础上,探讨了COⅠ条码序列在硬骨鱼类辅助物种鉴别和适用性,并对运用该条码序列库完善我国鱼类检索系统的可能途径进行了初步摸索。结果表明,COⅠ条码序列获取便捷,广泛适用硬骨鱼类物种鉴别,并可用于低级分类阶元的系统进化分析。本研究结果可为分子生物学辅助分类、市场监督、资源有序利用和保护提供参考。展开更多
文摘【目的】探明地方鸡种与引进鸡种遗传多态性特点,探讨COⅠ这一特定基因的特定区段作为DNA条形码在识别鸡种方面的可行性和有效性。【方法】以13个中国地方鸡种和2个国外引进品种为研究对象,利用DNA测序技术测定了线粒体细胞色素C氧化酶亚基Ⅰ(cytochrome c oxidaseⅠ,COⅠ)基因部分序列。【结果】选择的这段COⅠ基因序列有38个突变位点,15个鸡种单倍型多样度平均为0.963,核苷酸多样度平均为0.00518,其中引进鸡种明显低于地方鸡种(除藏鸡外);15个鸡种品种间Kimura双参数遗传距离为0.056%—0.917%,种内遗传距离为0—0.346%,15个鸡种的DNA分类和形态学分类基本一致,引进鸡种与地方鸡种分歧较远。【结论】利用线粒体细胞色素C氧化酶Ⅰ(COⅠ)这一特定基因的特定区段来做DNA条形编码的基础,进行不同鸡品种鉴定,具有可行性和有效性。
文摘利用mt DNA COⅠ基因片段(有效长度635 bp)对来自达里湖(DL)、岗更湖(GG)、呼伦湖(HL)、黑龙江(HLJ)、松花江(SHJ)和乌苏里江(WSL)6个地理群体共72个样品进行了遗传多样性和遗传结构分析。从碱基组成来看,6个地理群体在COⅠ区段上的A+T含量(52.37%)明显高于G+C含量(47.67%),符合脊椎动物的碱基组成特点;从各项遗传参数来看,达里湖流域2个群体(DL和GG)的遗传多样性低于黑龙江流域4个群体(HL、HLJ、SHJ、WSL);6个群体共检测到20个单倍型,并共享1个单倍型,达里湖流域群体共享1个单倍型,黑龙江流域群体共享1个单倍型。虽然达里湖流域和黑龙江流域各群体间遗传分化明显(P<0.05),遗传结构存在差异,但AMOVA检验不支持这种遗传差异;另外,群体间频繁的基因交流进一步证实,6个地理群体亲缘关系近,有共同的起源,尚未达到分化为不同的种或亚种的水平。
文摘沙棘木蠹蛾(Holcocerus hippophaecolust)和榆木蠹蛾(H.vicarious)为木蠹蛾科(Cossidae)线角木蠹蛾属(Holcocerus)的近缘种,二者形态相似,难于进行区分。本文基于线粒体COⅠ、COⅡ和Cyt b基因片段,对我国不同种群的沙棘木蠹蛾和榆木蠹蛾进行遗传差异分析,以探讨二者之间的亲缘关系。遗传距离分析表明,沙棘木蠹蛾和榆木蠹蛾在COⅠ、COⅡ和Cyt b 3个片段的遗传差异分别为0.009,0.001和0.062,仅相当于二者种内遗传差异均值范畴,远远低于同属的种间遗传距离均值,表明两种昆虫之间存在高度的遗传相似性。两种木蠹蛾在3个基因位点共检测到6个共享单倍型,占单倍型总数的四分之一,且共享单倍型分布不受地理距离限制,显示两个种的种群之间尚未完全分化。系统发育树分析中,沙棘木蠹蛾和榆木蠹蛾在3个基因序列均未构成独立分支,而是形成了混合集群。介于沙棘木蠹蛾和榆木蠹蛾之间极高的遗传相似性,对二者之间的亲缘进化关系进行了讨论。
文摘【目的】研究贵州省茶棍蓟马不同地理种群间的遗传多样性,为掌握贵州省茶棍蓟马的遗传动态和扩散规律及制定防控措施提供理论依据。【方法】以贵州省11个不同地理区域的茶棍蓟马种群mtDNA CO Ⅰ和mtDNA CO Ⅱ基因序列为靶标,利用MEGA 6.0、DnaSP 5.10、Arlequin 3.5.2.2和Network 2.0等软件对种群遗传分化、基因流水平、分子变异及种群遗传多样性等进行分析。【结果】基于mtDNA CO Ⅰ和mtDNA CO Ⅱ基因分析时,分别检测到11种和9种单倍型,各地理种群的单倍型多样度(Hd)较高,分别为0.609和0.633;总体遗传固定指数(FST)分别为0.11902和0.09052,基因流(Nm)分别为2.00和2.51,表明贵州省茶棍蓟马各地理种群间的基因交流水平较高,种群间遗传分化较小。mtDNA CO Ⅰ和mtDNA CO Ⅱ基因序列的单倍型网状树分析均无明显分支;种群间的分子变异(AMOVA)分析结果表明,茶棍蓟马的遗传变异主要来自种群内部。总群体的中性检验Fu’s Fs不显著(P>0.05),且错配分布曲线呈现多峰,说明贵州省茶棍蓟马种群在较近的历史时期内保持相对稳定,未经历明显的种群扩张,或处于临界状态。【结论】贵州省茶棍蓟马种群遗传多样性较丰富,虽然尚未形成明显的种群扩张,但可能已处于临界状态,应积极采取综合防控措施,力争将茶棍蓟马种群数量控制在危害水平以下,以保证贵州省茶产业生产健康可持续发展。
文摘为了揭示吉林山鸡细胞色素C氧化酶亚基Ⅰ(COⅠ)基因的遗传多样性及其与地方鸡种的遗传关系,试验采用DNA测序技术测定吉林山鸡线粒体COⅠ基因序列。结果表明:20只山鸡样本mt DNA COⅠ基因序列中,共检测到核苷酸多态位点19个,20个个体具有4种单倍型,吉林山鸡群体内平均核苷酸多样度(Pi)为0.002 88,单倍型多样性(Hd)为0.625,可以作为品种鉴定的依据。吉林山鸡种群的遗传多样性水平较低,建议对吉林山鸡种群的保护制订更有效的繁殖计划,避免近亲杂交。
基金Fund on the Special Fund for Agro-scientific Research in the Public Interest(No.201203065)Special Fund for Ecological Restoration in Adjacent Waters of Grass Sand Reservoir
文摘To determine the role of mitochondrial COⅠgene in classification and identification of species,a total of 39 single individuals from nine species pertaining to two genera of Cynoglossidae in the Yangtze Estuary were barcoded by COⅠ,sequenced and compared with that of other Cynoglossidae species recorded in the Gen Bank. Total genomic DNA was extracted from each scale sample using the classic phenol / chloroform extraction method. Six hundred and fifty base pairs( bp)COⅠfragments were amplified using the primers ’i. e. ’ F1: 5’- TCA ACC AAC CAC AAA GAC ATT GGC AC- 3’,R1: 5’- TAG ACT TCT GGG TGG CCA AAG AAT CA- 3’. Every PCR amplification was performed in a total volume of 50 μL of PCR mixture. PCR products were purified and then sequenced in both forward and reverse directions using an ABI PRISMTM 3730 XL Automated Sequencer. DNA sequences were aligned with clustal W using default parameters. Base composition,variable and parsimony informative sites were determined using MEGA 5. 0. Neighbor- joining( NJ) and Maximum parsimony( MP) phylogenetic trees were constructed for COⅠhaplotypes( Kimura 2 Parameter substitution model,K2P; 1 000 bootstraps pseudoreplications) using MEGA 5. 0. Using the MEGA5. 0 software for statistical analysis,the averaged AT content was greater than the GC content( Tab. 2). The GC content of codon position 1 averaged 53. 8%( 51. 8%-57. 3%),which of position 2 for 42. 0%,and that of position 3 ranged from 28. 1% to 37. 8% in average of 32. 4%( Tab. 4). The transitional pairs( si) was slightly more than the transversional pairs( sv),and the ratio( R = si/sv) was 1. 45( Tab. 3). Analysis of the frequency of amino acids in COⅠgene encoding protein showed that the highest frequency of amino acid was leucine,and the lowest frequency of amino acid was tryptophan( Tab. 5). The average K2 P distances pairwise-species and within-species were 0. 191 and 0. 003,respectively( Tab. 6). The K2 P distance pairwise-species was 63. 7 times of that within-species. According to the MP and NJ trees for all 39 sequences,it demonstrated that Cynoglossinae in the Yangtze Estuary was a monophyletic group( Fig. 1). However,the phylogenetic relationships revealed by the COⅠsequence analyses were not consistent with those inferred from morphological classification. In contrast to morphological classification,Paraplagusis japonica of Rhinoplagusia and Cynoglossus robustus of Cynoglossoides were placed in a same clade with support of a high bootstrap value( Fig. 1). The species in the sub-genera of Areliscus clustered on an independent branch. But synonymic phenomena existed in the following two groups of species: C. abbreviatus and C. purpureomaculatus,C. lighti and C. joyner,with the pairwise-species of 0. 002 and 0. 007,respectively( Tab. 6). Our results highlight that the information from COⅠsequences not only can filter out the synonym of the same species,but also be able to be used to carry out effective identification for Cynoglossidae species,which further shows that mitochondrial COⅠis feasible as the classification barcode.
文摘运用一对特异性引物扩增并测定了南海硬骨鱼类40个物种89个样本的线粒体DNA的细胞色素c氧化酶Ⅰ(COⅠ)约660bp的部分序列,即COⅠ条码序列。综合比较了《中国鱼类系统检索》、《海洋鱼类志》、FishBase鱼类形态学分类库、ITIS综合分类学信息系统和生物条码系统(The Barcode of Life Data System,BOLD)的相应形态学与DNA序列资料,在揭示完善我国鱼类检索系统必要性基础上,探讨了COⅠ条码序列在硬骨鱼类辅助物种鉴别和适用性,并对运用该条码序列库完善我国鱼类检索系统的可能途径进行了初步摸索。结果表明,COⅠ条码序列获取便捷,广泛适用硬骨鱼类物种鉴别,并可用于低级分类阶元的系统进化分析。本研究结果可为分子生物学辅助分类、市场监督、资源有序利用和保护提供参考。
