秦巴山区3种步甲（鞘翅目：步甲科）种群遗传结构和种群数量变化历史的研究
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摘要
秦岭-巴山地区是中国生物多样性起源和演化中心之一,分布着丰富的动植物资源,古北界和东洋界种类在这里交汇分布,造就了独特的生物区系组成。该地区如同北半球其他地区一样,在第四纪也经历过多次冰期-间冰期回旋,这一气候事件往往会在物种种群遗传多样性的地理分布上留下痕迹。了解该地区动物遗传结构、多样性及古冰川作用对该地区动物地理分布格局的影响对动物资源保护和农林业合理布局具有重要意义。步甲科昆虫物种古老,种群内遗传分化明显,大多数步甲科昆虫是重要的天敌昆虫,对农林业害虫的控制起到关键性作用,了解其在分布区的遗传多样性状况和分布模式对于物种的保护和病虫害防治有一定的理论指导意义。
本论文利用线粒体细胞色素氧化酶Ⅰ基因和细胞色素氧化酶Ⅱ基因长为1601-1602 bp的连续DNA片段(cox1-cox2)作为分子标记,分别对广泛分布于该区的耶屁步甲(Pheropsophus jessoensis)、赤胸疏爪步甲(Dolichus halensis (Schaller 1783))和淡足青步甲(Chlaenius pallies)进行了种群遗传结构和种群数量动态历史进行了分析,研究结果如下：
1.获得25个居群的184个耶屁步甲个体标记序列,该序列长1601 bp,其中包括879 bp的线粒体CoⅠ基因、633 bp的线粒体CoⅡ基因和69 bp的tRNA-Leu间隔区。共有82个多态性位点,包括43个简约信息位点、38个单变异位点和1个碱基缺失位点,检测出92个单倍型。在tRNA-Leu间隔区中有1个碱基缺失现象。种群总的遗传多样性指数(HT=0.960)、居群内平均遗传多样性指数(HS=0.880)以及单倍型多样性(Hd=0.963)都很高,但核苷酸多样性(P,=0.0037)较低。居群独享单倍型占总单倍型数的79.34%。单倍型网络图和系统发育分析表明秦巴山区分布的耶屁步甲有8个单倍型进化枝(A-H)。AMOVA分析表明居群间和谱系枝间遗传分化显著(居群间：FST=0.078,p<0.01；谱系枝间：FST=0.558,p<0.01),种群遗传分化主要源于居群内和谱系枝间。但SAMOVA和PERMUTE分析表明不存在明显的谱系地理结构(NST=0.107>GST=0.083,p=0.075>0.05)。统计谱系地理学分析结果拒绝种群片段化模型假设。基因流分析显示居群间存在较为频繁的基因流。Mantel检验结果表明遗传分化和地理距离之间存在显著性关联(R=0.360,p=0.002<0.01),符合距离隔离模型(IBDM)的推断。错配分布、中性检验以及BSP分析结果显示该物种曾经经历过多次扩张,扩张时间大致为0.2-0.01 Myr,且在0.05-0.01 Myr发生了一次快速扩散。利用BEAST软件,估算出秦巴山区该种群的溯祖时间为0.2994(95%HPD：0.1971-0.5034)Myr。
2.获得24个居群的191个赤胸疏爪步甲个体标记序列,该序列长1 601 bp,其中包括882 bp的线粒体CoⅠ基因、656 bp的线粒体CoⅡ基因和63 bp的tRNA-Leu间隔区,无碱基缺失或插入现象。共发现45个多态性位点,包括15个简约信息位点和30个单变异位点,检测出53个单倍型。种群总的遗传多样性指数(HT=0.705)、居群内平均遗传多样性指数(HS=0.612)以及单倍型多样性(Hd=0.769)都较高,但核苷酸多样性(Pi=0.0033)较低。居群独享单倍型占总单倍型数的65.71%。单倍型网络图和系统发育分析表明秦巴山区分布的赤胸疏爪步甲有2个单倍型进化枝(A-B)。AMOVA分析表明遗传分化在种群间和进化枝间很显著(居群间FST=0.134,p<0.01；谱系枝间FST=0.433,p<0.01),种群遗传分化同样主要源于居群内和进化枝之间。同时,SAMOVA和PERMUTE分析表明不存在明显的谱系地理结构(NST=0.154>GST=0.132,p=0.085>0.05)。统计谱系地理学分析结果接受种群片段化模型假设。基因流分析显示居群间存在较为频繁的基因流,但Mantel检验发现遗传分化和地理距离之间不存在显著性关联(R=0.055,p=0.679>0.05),拒绝距离隔离模型(IBDM)的推断。错配分布、中性检验以及BSP分析结果都表明该物种曾经经历过多次的扩张,发生时间大致为0.05-0.01 Myr,且这一期间为快速扩散时间。利用BEAST软件,估算出秦巴山区该物种的溯祖时间为0.2609 (95%HPD:0.1490～0.4300) Myr。
3.获得13个居群的151个淡足青步甲个体标记序列,该序列长1602 bp,其中包括882 bp的线粒体CoⅡ基因、658 bp的线粒体CoⅡ基因和62 bp的tRNA-Leu间隔区,其中在tRNA-Leu司隔区中发现了1个碱基插入现象。共发现57个多态性位点,包括45个简约信息位点、11个单变异位点和一个碱基插入位点,检测出65个单倍型。种群总的遗传多样性指数(HT=0.985)、居群内平均遗传多样性指数(HS=0.885)以及单倍型多样性(Hd=0.972)都很高,但核苷酸多样(Pi=0.0025)较低。居群独享单倍型占总单倍型数的73.85%。单倍型网络图和系统发育分析表明秦巴山区分布的淡足青步甲有3个单倍型谱系枝(A-C)。AMOVA分析表明遗传分化在种群间和谱系枝间很显著(居群间FST=0.079,p<0.01；谱系枝间FST=0.338,p<0.01),种群遗传分化主要源于居群内和谱系枝之间。同时,SAMOVA和PERMUTE分析表明不存在明显的谱系地理结构(NST=0.114>GST=0.101,p=0.33>0.05)。统计谱系地理学分析结果拒绝种群片段化模型假设。基因流分析结果显示居群间存在较为频繁的基因流,但Mantel检验发现遗传分化和地理距离之间不存在显著性关联(R=0.126,p=0.855>0.05),拒绝距离隔离模型(IBDM)的推断。错配分布、中性检验以及BSP分析结果都表明该物种曾经经历过多次扩张,扩张时间大致为0.125-0.05 Myr,且在0.1-0.075 Myr发生了一次快速扩散,淡足青步甲发生快速扩散的时间早于前面两个物种。利用BEAST软件,推算秦巴山区淡足青步甲溯祖时间为0.2544 (95% HPD:0.1594～0.3699) Myr。
综上所述,本文所研究的秦巴山区三种步甲种群遗传分化明显,每一物种居群间存在较强的基因交流,使得种群内不具有明显的谱系地理结构。受第四纪冰期造成的环境剧烈变化以及冰期与间冰期的反复交替的影响,此三种步甲均发生过多次种群扩张。认为瓶颈效应、冰期后的扩张以及可能的长距迁移是步甲形成现如今遗传分布格局的主要因素。
The Tsinling-Dabashan Mountains are one of centers for the origin and evolution of biodiversity in China. These areas contain rich animal and plant resources. The Oriental and Palaearctic species congregate here forming a specific biotic province. The past climatic events, such as the Quaternary glaciation. could leave vestiges in geographical distribution of genetic diversity of population. Like other regions in the northern hemisphere, the Tsinling-Dabashan Mountains also experienced several glacial-interglacial cycles. Understanding the pattern of genetic structure and diversity condition of fauna are distributing on Tsin-ba and the effects of palaeoglaciation on geographical distribution pattern have great value. The species belonging to Carabidea family are ancient, and the most of them are beneficial insect with significial genetic differentiation. They play an important role in pest management. Complement understanding about the conditions of genetic diversity and distribution pattern of beetles on the Tsin-ba Mountains is important for species protection and also provides the foundation for pest control.
In this study, mtDNA coxl-cox2 were emplyed to reveal the populations structure and historical demography of Pheropsophus jessoensis, Dolichus halensis (Schaller.1783) and Chlaenius pallies. All of them are widespread species. The results of investigations are offered as follows:
1. A total of 184 adults of the ground beetle P. jessoensis were collected from 25 locations on the Tsinling-Dabashan Mountains. In length of 1601bp fragment, a total of 879 bp were sequenced for the mtCoI gene, and 653 bp for the mtCoII gene. The intervening tRNA Leu gene was 69 bp in length, which contained a 1-bp deletion.81 positions were polymorphic sites, composed of 43 parsimony informative sites and 38 singleton variable sites. These polymorphic sites identified 92 haplotypes. The total genetic diversity HT (0.960) among all sampled populations, the average within-population diversity Hs (0.880) and the haplotype diversity Hd (0.963) of total population were considerable high, but the nucleotide diversity Pi (0.0037) of total population was fairly low. And 79.34% of total haplotypes are private haplotypes with lower frequency. Phylogenetic analysis (Bayesian inference) and parsimonious network of the 92 haplotypes revealed eight major clades (A-H). AMOVA analysis suggested that most of the variation was attributed to within populations and among clades and the genetic differentiation among populations and among clades were significiant (among population:FST=0.078, p<0.01; among phylogroups:FST=0.558, p<0.01). But the SAMOVA tests and PERMUTE analyses failed to reveal any meaningful phylogeographic structure (NST=0.107> GST=0.083, p=0.075>0.05). The results of statistical phylogeography analysis rejected the model of population fragmentation. The gene flow calculated through the Nm was high between many pairs of populations, and Mantel test results showed significant correlation between the pairwise calculated genetic distance and pairwise calculated geographical distance of the populations (R= 0.360, p=0.002< 0.01), indicating the presence of isolation-by-distance model (IBDM). The results of neutral test and mismatch distribution analyses of total population and the eight mtDNA clades suggest several demographic expansions between 0.2Myr to 0.01 Myr, and a sudden expansion was identified between 0.05 Myr to 0.01 Myr by BSP analysis. The estimated age of the origin of the P. jessoensis was 0.30 Myr, with a 95% interval of confidence between 0.20 and 0.50 Myr by BEAST.
2. A total of 191 adults of the ground beetle D.halensis were collected from 24 locations on the Tsinling-Dabashan Mountains. In length of 1601bp fragment, a total of 882 bp were sequenced for the mtCol gene, and 656 bp for the mtCoⅡgene. The intervening tRNA Leu gene was 63 bp in length.45 positions were polymorphic sites, composed of 15 parsimony informative sites and 30 singleton variable sites. These polymorphic sites identified 53 haplotypes. The total genetic diversity HT (0.705) among all sampled populations, the average within-population diversity HS (0.612) and the haplotype diversity Hd (0.769) of total population were considerable high, but the nucleotide diversity Pi (0.0033) of total population was fairly low. And 65.71% of total haplotypes are private haplotypes with lower frequency. Phylogenetic analysis (Bayesian inference) and parsimonious network of the 53 haplotypes revealed 2 major clades (A-B). AMOVA analysis suggested that most of the variation was attributed to within populations and among clades and the genetic differentiation among populations and among clades were significiant (among population:FST=0.134, p<0.01; among clades:FST=0.433,p<0.01). But the SAMOVA tests and PERMUTE analyses failed to reveal any meaningful phylogeographic structure (NST=0.154> GST=0.132,p= 0.085>0.05). The results of statistical phylogeography analysis accepted the model of population fragmentation. The gene flow calculated through the Nm was high between many pairs of populations, but Mantel test results showed no significant correlation between the pairwise calculated genetic distance and pairwise calculated geographical distance of the populations (R=0.055, p=0.679> 0.05). rejecting the supposed of isolation-by-distance model (IBDM). The results of neutral test and mismatch distribution analyses of total population and the two mtDNA haplogroups suggest several demographic expansion between 0.05 Myr to 0.01 Myr, and a sudden expansion was identified between 0.05 Myr to 0.01 Myr by BSP analysis. The tMRCA of all D.halensis was date to 0.2609 Myr before present (95% of the highest posterior density [HPD]:0.4300 to 0.1490 Myr).
3. A total of 151 adults of the ground beetle C.pallipes were collected from 13 locations on the Tsinling-Dabashan Mountains. In length of 1602bp fragment, a total of 882 bp were sequenced for the mtCoⅠgene, and 658 bp for the mtCoⅡgene. The intervening tRNA Leu gene was 62 bp in length, which contained a 1-bp insertion.56 positions were polymorphic sites, composed of 45 parsimony informative sites and 11 singleton variable sites. These polymorphic sites identified 65 haplotypes. The total genetic diversity HT (0.985) among all sampled populations, the average within-population diversity HS (0.885) and the haplotype diversity Hd (0.972) of total population were considerable high, but the nucleotide diversity Pi (0.0025) of total population was fairly low. And 73.85% of total haplotypes are private haplotypes with lower frequency. Phylogenetic analysis (Bayesian inference) and parsimonious network of the 92 haplotypes revealed three major haplogroups (A-C). AMOVA analysis suggested that most of the variation was attributed to within populations and among haplogroups and the genetic differentiation among populations and among phylogroups were significiant (among population:FST=0.079,p<0.01; among phylogroups:FST= 0.338, p<0.01). But the SAMOVA tests and PERMUTE analyses failed to reveal any meaningful phylogeographic structure (NST=0.114> GST=0.101, p=0.33>0.05), The results of statistical phylogeography analysis rejected the model of population fragmentation. The gene flow calculated through the Nm was high between many pairs of populations, and Mantel test results showed no significant correlation between the pairwise calculated genetic distance and pairwise calculated geographical distance of the populations (R=0.126, p=0.855>0.05), rejecting the supposed of isolation-by-distance model (IBDM). The results of neutral test and mismatch distribution analyses of total population and the three mtDNA haplogroups suggest several demographic expansions between 0.125 Myr to 0.05 Myr. and a sudden expansion was identified between 0.075 Myr to 0.1 Myr by BSP analysis. The estimated age of the origin of the C.pallipes was 0.2544 Myr, with a 95% interval of confidence between 0.1594 and 0.3699 Myr by BEAST.
In conclusion, we consider that there are significant genetic differentiations between populations of beetles distributing on the Tsinling-Dabashan Mountains, but we failed to reveal any meaningful phylogeographic structure for the gene flow calculated through the Nm was high between many pairs of populations. Climate change by glacial-interglacial cycles in the Quaternary glaciation led to several demographic expansions of beetles. Bottleneck effect, demographic expansion after glaciation and long-distance migration are important factor for the pattern of genetic.
本论文利用线粒体细胞色素氧化酶Ⅰ基因和细胞色素氧化酶Ⅱ基因长为1601-1602 bp的连续DNA片段(cox1-cox2)作为分子标记,分别对广泛分布于该区的耶屁步甲(Pheropsophus jessoensis)、赤胸疏爪步甲(Dolichus halensis (Schaller 1783))和淡足青步甲(Chlaenius pallies)进行了种群遗传结构和种群数量动态历史进行了分析,研究结果如下：
1.获得25个居群的184个耶屁步甲个体标记序列,该序列长1601 bp,其中包括879 bp的线粒体CoⅠ基因、633 bp的线粒体CoⅡ基因和69 bp的tRNA-Leu间隔区。共有82个多态性位点,包括43个简约信息位点、38个单变异位点和1个碱基缺失位点,检测出92个单倍型。在tRNA-Leu间隔区中有1个碱基缺失现象。种群总的遗传多样性指数(HT=0.960)、居群内平均遗传多样性指数(HS=0.880)以及单倍型多样性(Hd=0.963)都很高,但核苷酸多样性(P,=0.0037)较低。居群独享单倍型占总单倍型数的79.34%。单倍型网络图和系统发育分析表明秦巴山区分布的耶屁步甲有8个单倍型进化枝(A-H)。AMOVA分析表明居群间和谱系枝间遗传分化显著(居群间：FST=0.078,p<0.01；谱系枝间：FST=0.558,p<0.01),种群遗传分化主要源于居群内和谱系枝间。但SAMOVA和PERMUTE分析表明不存在明显的谱系地理结构(NST=0.107>GST=0.083,p=0.075>0.05)。统计谱系地理学分析结果拒绝种群片段化模型假设。基因流分析显示居群间存在较为频繁的基因流。Mantel检验结果表明遗传分化和地理距离之间存在显著性关联(R=0.360,p=0.002<0.01),符合距离隔离模型(IBDM)的推断。错配分布、中性检验以及BSP分析结果显示该物种曾经经历过多次扩张,扩张时间大致为0.2-0.01 Myr,且在0.05-0.01 Myr发生了一次快速扩散。利用BEAST软件,估算出秦巴山区该种群的溯祖时间为0.2994(95%HPD：0.1971-0.5034)Myr。
2.获得24个居群的191个赤胸疏爪步甲个体标记序列,该序列长1 601 bp,其中包括882 bp的线粒体CoⅠ基因、656 bp的线粒体CoⅡ基因和63 bp的tRNA-Leu间隔区,无碱基缺失或插入现象。共发现45个多态性位点,包括15个简约信息位点和30个单变异位点,检测出53个单倍型。种群总的遗传多样性指数(HT=0.705)、居群内平均遗传多样性指数(HS=0.612)以及单倍型多样性(Hd=0.769)都较高,但核苷酸多样性(Pi=0.0033)较低。居群独享单倍型占总单倍型数的65.71%。单倍型网络图和系统发育分析表明秦巴山区分布的赤胸疏爪步甲有2个单倍型进化枝(A-B)。AMOVA分析表明遗传分化在种群间和进化枝间很显著(居群间FST=0.134,p<0.01；谱系枝间FST=0.433,p<0.01),种群遗传分化同样主要源于居群内和进化枝之间。同时,SAMOVA和PERMUTE分析表明不存在明显的谱系地理结构(NST=0.154>GST=0.132,p=0.085>0.05)。统计谱系地理学分析结果接受种群片段化模型假设。基因流分析显示居群间存在较为频繁的基因流,但Mantel检验发现遗传分化和地理距离之间不存在显著性关联(R=0.055,p=0.679>0.05),拒绝距离隔离模型(IBDM)的推断。错配分布、中性检验以及BSP分析结果都表明该物种曾经经历过多次的扩张,发生时间大致为0.05-0.01 Myr,且这一期间为快速扩散时间。利用BEAST软件,估算出秦巴山区该物种的溯祖时间为0.2609 (95%HPD:0.1490～0.4300) Myr。
3.获得13个居群的151个淡足青步甲个体标记序列,该序列长1602 bp,其中包括882 bp的线粒体CoⅡ基因、658 bp的线粒体CoⅡ基因和62 bp的tRNA-Leu间隔区,其中在tRNA-Leu司隔区中发现了1个碱基插入现象。共发现57个多态性位点,包括45个简约信息位点、11个单变异位点和一个碱基插入位点,检测出65个单倍型。种群总的遗传多样性指数(HT=0.985)、居群内平均遗传多样性指数(HS=0.885)以及单倍型多样性(Hd=0.972)都很高,但核苷酸多样(Pi=0.0025)较低。居群独享单倍型占总单倍型数的73.85%。单倍型网络图和系统发育分析表明秦巴山区分布的淡足青步甲有3个单倍型谱系枝(A-C)。AMOVA分析表明遗传分化在种群间和谱系枝间很显著(居群间FST=0.079,p<0.01；谱系枝间FST=0.338,p<0.01),种群遗传分化主要源于居群内和谱系枝之间。同时,SAMOVA和PERMUTE分析表明不存在明显的谱系地理结构(NST=0.114>GST=0.101,p=0.33>0.05)。统计谱系地理学分析结果拒绝种群片段化模型假设。基因流分析结果显示居群间存在较为频繁的基因流,但Mantel检验发现遗传分化和地理距离之间不存在显著性关联(R=0.126,p=0.855>0.05),拒绝距离隔离模型(IBDM)的推断。错配分布、中性检验以及BSP分析结果都表明该物种曾经经历过多次扩张,扩张时间大致为0.125-0.05 Myr,且在0.1-0.075 Myr发生了一次快速扩散,淡足青步甲发生快速扩散的时间早于前面两个物种。利用BEAST软件,推算秦巴山区淡足青步甲溯祖时间为0.2544 (95% HPD:0.1594～0.3699) Myr。
综上所述,本文所研究的秦巴山区三种步甲种群遗传分化明显,每一物种居群间存在较强的基因交流,使得种群内不具有明显的谱系地理结构。受第四纪冰期造成的环境剧烈变化以及冰期与间冰期的反复交替的影响,此三种步甲均发生过多次种群扩张。认为瓶颈效应、冰期后的扩张以及可能的长距迁移是步甲形成现如今遗传分布格局的主要因素。
The Tsinling-Dabashan Mountains are one of centers for the origin and evolution of biodiversity in China. These areas contain rich animal and plant resources. The Oriental and Palaearctic species congregate here forming a specific biotic province. The past climatic events, such as the Quaternary glaciation. could leave vestiges in geographical distribution of genetic diversity of population. Like other regions in the northern hemisphere, the Tsinling-Dabashan Mountains also experienced several glacial-interglacial cycles. Understanding the pattern of genetic structure and diversity condition of fauna are distributing on Tsin-ba and the effects of palaeoglaciation on geographical distribution pattern have great value. The species belonging to Carabidea family are ancient, and the most of them are beneficial insect with significial genetic differentiation. They play an important role in pest management. Complement understanding about the conditions of genetic diversity and distribution pattern of beetles on the Tsin-ba Mountains is important for species protection and also provides the foundation for pest control.
In this study, mtDNA coxl-cox2 were emplyed to reveal the populations structure and historical demography of Pheropsophus jessoensis, Dolichus halensis (Schaller.1783) and Chlaenius pallies. All of them are widespread species. The results of investigations are offered as follows:
1. A total of 184 adults of the ground beetle P. jessoensis were collected from 25 locations on the Tsinling-Dabashan Mountains. In length of 1601bp fragment, a total of 879 bp were sequenced for the mtCoI gene, and 653 bp for the mtCoII gene. The intervening tRNA Leu gene was 69 bp in length, which contained a 1-bp deletion.81 positions were polymorphic sites, composed of 43 parsimony informative sites and 38 singleton variable sites. These polymorphic sites identified 92 haplotypes. The total genetic diversity HT (0.960) among all sampled populations, the average within-population diversity Hs (0.880) and the haplotype diversity Hd (0.963) of total population were considerable high, but the nucleotide diversity Pi (0.0037) of total population was fairly low. And 79.34% of total haplotypes are private haplotypes with lower frequency. Phylogenetic analysis (Bayesian inference) and parsimonious network of the 92 haplotypes revealed eight major clades (A-H). AMOVA analysis suggested that most of the variation was attributed to within populations and among clades and the genetic differentiation among populations and among clades were significiant (among population:FST=0.078, p<0.01; among phylogroups:FST=0.558, p<0.01). But the SAMOVA tests and PERMUTE analyses failed to reveal any meaningful phylogeographic structure (NST=0.107> GST=0.083, p=0.075>0.05). The results of statistical phylogeography analysis rejected the model of population fragmentation. The gene flow calculated through the Nm was high between many pairs of populations, and Mantel test results showed significant correlation between the pairwise calculated genetic distance and pairwise calculated geographical distance of the populations (R= 0.360, p=0.002< 0.01), indicating the presence of isolation-by-distance model (IBDM). The results of neutral test and mismatch distribution analyses of total population and the eight mtDNA clades suggest several demographic expansions between 0.2Myr to 0.01 Myr, and a sudden expansion was identified between 0.05 Myr to 0.01 Myr by BSP analysis. The estimated age of the origin of the P. jessoensis was 0.30 Myr, with a 95% interval of confidence between 0.20 and 0.50 Myr by BEAST.
2. A total of 191 adults of the ground beetle D.halensis were collected from 24 locations on the Tsinling-Dabashan Mountains. In length of 1601bp fragment, a total of 882 bp were sequenced for the mtCol gene, and 656 bp for the mtCoⅡgene. The intervening tRNA Leu gene was 63 bp in length.45 positions were polymorphic sites, composed of 15 parsimony informative sites and 30 singleton variable sites. These polymorphic sites identified 53 haplotypes. The total genetic diversity HT (0.705) among all sampled populations, the average within-population diversity HS (0.612) and the haplotype diversity Hd (0.769) of total population were considerable high, but the nucleotide diversity Pi (0.0033) of total population was fairly low. And 65.71% of total haplotypes are private haplotypes with lower frequency. Phylogenetic analysis (Bayesian inference) and parsimonious network of the 53 haplotypes revealed 2 major clades (A-B). AMOVA analysis suggested that most of the variation was attributed to within populations and among clades and the genetic differentiation among populations and among clades were significiant (among population:FST=0.134, p<0.01; among clades:FST=0.433,p<0.01). But the SAMOVA tests and PERMUTE analyses failed to reveal any meaningful phylogeographic structure (NST=0.154> GST=0.132,p= 0.085>0.05). The results of statistical phylogeography analysis accepted the model of population fragmentation. The gene flow calculated through the Nm was high between many pairs of populations, but Mantel test results showed no significant correlation between the pairwise calculated genetic distance and pairwise calculated geographical distance of the populations (R=0.055, p=0.679> 0.05). rejecting the supposed of isolation-by-distance model (IBDM). The results of neutral test and mismatch distribution analyses of total population and the two mtDNA haplogroups suggest several demographic expansion between 0.05 Myr to 0.01 Myr, and a sudden expansion was identified between 0.05 Myr to 0.01 Myr by BSP analysis. The tMRCA of all D.halensis was date to 0.2609 Myr before present (95% of the highest posterior density [HPD]:0.4300 to 0.1490 Myr).
3. A total of 151 adults of the ground beetle C.pallipes were collected from 13 locations on the Tsinling-Dabashan Mountains. In length of 1602bp fragment, a total of 882 bp were sequenced for the mtCoⅠgene, and 658 bp for the mtCoⅡgene. The intervening tRNA Leu gene was 62 bp in length, which contained a 1-bp insertion.56 positions were polymorphic sites, composed of 45 parsimony informative sites and 11 singleton variable sites. These polymorphic sites identified 65 haplotypes. The total genetic diversity HT (0.985) among all sampled populations, the average within-population diversity HS (0.885) and the haplotype diversity Hd (0.972) of total population were considerable high, but the nucleotide diversity Pi (0.0025) of total population was fairly low. And 73.85% of total haplotypes are private haplotypes with lower frequency. Phylogenetic analysis (Bayesian inference) and parsimonious network of the 92 haplotypes revealed three major haplogroups (A-C). AMOVA analysis suggested that most of the variation was attributed to within populations and among haplogroups and the genetic differentiation among populations and among phylogroups were significiant (among population:FST=0.079,p<0.01; among phylogroups:FST= 0.338, p<0.01). But the SAMOVA tests and PERMUTE analyses failed to reveal any meaningful phylogeographic structure (NST=0.114> GST=0.101, p=0.33>0.05), The results of statistical phylogeography analysis rejected the model of population fragmentation. The gene flow calculated through the Nm was high between many pairs of populations, and Mantel test results showed no significant correlation between the pairwise calculated genetic distance and pairwise calculated geographical distance of the populations (R=0.126, p=0.855>0.05), rejecting the supposed of isolation-by-distance model (IBDM). The results of neutral test and mismatch distribution analyses of total population and the three mtDNA haplogroups suggest several demographic expansions between 0.125 Myr to 0.05 Myr. and a sudden expansion was identified between 0.075 Myr to 0.1 Myr by BSP analysis. The estimated age of the origin of the C.pallipes was 0.2544 Myr, with a 95% interval of confidence between 0.1594 and 0.3699 Myr by BEAST.
In conclusion, we consider that there are significant genetic differentiations between populations of beetles distributing on the Tsinling-Dabashan Mountains, but we failed to reveal any meaningful phylogeographic structure for the gene flow calculated through the Nm was high between many pairs of populations. Climate change by glacial-interglacial cycles in the Quaternary glaciation led to several demographic expansions of beetles. Bottleneck effect, demographic expansion after glaciation and long-distance migration are important factor for the pattern of genetic.
引文
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