Genetic variation in skipjack tuna Katsuwonus pelamis(L.) using PCR‐RFLP analysis of the mitochondrial DNA D‐loop region

A high level of genetic diversity was observed in Katsuwonus pelamis populations from India ( h  = 0·952, ne  = 14·3) and Japan ( h  = 0·897, ne  = 8·9). The log‐likelihood ( G )‐based exact test revealed significant heterogeneity in the distribution of haplotypes between the two populations ( P  < 0·01, s . e . = 0·001). This result suggests that the two populations should now be treated as demographically independent and managed separately.     

秦岭细鳞鲑群体遗传结构

秦岭细鳞鲑(Brachymystax lenok tsinlingensis)是秦岭地区特有鱼类,近年来由于环境恶化及人类活动的加剧,已对其造成严重影响,种群处于濒危状态,因此研究秦岭细鳞鲑的群体遗传结构、演化历史、分布动态等对其进行有效保护具有重要意义。本研究用线粒体D-loop区序列对秦岭地区6个群体(n=112)进行了遗传结构和群体演化分析。D-loop区扩增出的891bp序列在112个个体中,检测到42个变异位点,共26个单倍型;碱基序列总的单倍型多样度较高为0.883,核苷酸多样度为0.00799。AMOVA分析显示,60.05%的分子差异位于群体内,39.95%的分子差异位于群体间,Fst值统计检验表明,除那布大河群体与漳河群体和千河群体之间差异不显著之外,其余两两群体之间Fst值统计检验均为显著。系统树和单倍型网络图分析表明,6个地理群体的单倍型按照渭河上游和渭河中游两个河段形成两个大的类群,且5个群体共享一个单倍型H2,表明这些群体具有相同的演化历史,为同一个祖先群体演化而来。中性检验和歧点分布显示,秦岭细鳞鲑种群大小保持相对稳定,未经历明显的种群扩张。同时建议将渭河上游秦岭细鳞鲑群体作为一个整体进行重点保护。     

Origin of rabbit (Oryctolagus cuniculus) in China: evidence from mitochondrial DNA control region sequence analysis

A fragment of mitochondrial DNA (mtDNA) control region (approximately 700 bp) was sequenced in 104 individuals from 20 breeds (three Chinese domestic breeds, five recently derived breeds and 12 introduced breeds) of domestic rabbits, Oryctolagus cuniculus. Nineteen sites were polymorphic, with 18 transitions and one insertion/deletion, and eight haplotypes (A1, A2, A3, A4, A5, A6, A7 and A8) were identified. Haplotype A1 was the most common and occurred in 89 individuals. In the 25 Chinese rabbits, only haplotype A1 was observed, while four haplotypes (A1, A3, A5 and A6) were found in 26 recently derived individuals. Haplotype A2 was shared by seven individuals among three introduced strains. The other six haplotypes accounted for 0.96-1.92% of the animals. Combined with the published sequences of European rabbits, a reduced median-joining network was constructed. The Chinese rabbit mtDNAs were scattered into two clusters of European rabbits. These results suggest that the (so-called) Chinese rabbits were introduced from Europe. Genetic diversity in Chinese rabbits was very low.     

Polymorphic sequence in the D-loop region of equine mitochondrial DNA

The D-loop regions in equine mitochondrial DNA were cloned from three thoroughbred horses by polymerase chain reaction (PCR). The total number of bases in the D-loop region were 1114bp, 1115bp and 1146bp. The equine D-loop region is A/T rich like many other mammalian D-loops. The large central conserved sequence block and small conserved sequence blocks 1, 2 and 3, that are common to other mammals, were observed. Between conserved sequence blocks 1 and 2 there were tandem repeats of an 8bp equine-specific sequence TGTGCACC, and the number of tandem repeats differed among individual horses. The base composition in the unit of these repeats is G/C rich as are the short repeats in the D-loops of rabbit and pig. Comparing DNA sequences between horse and other mammals, the difference in the D-loop region length is mostly due to the difference in the number of DNA sequences at both extremities. The similarities of the DNA sequences are in the middle part of the D-loop. In comparison of the sequences among three thoroughbred horses, it was determined that the region between tRNA^Pro and the large central conserved sequence block was the richest in variation. PCR primers in the D-loop region were designed and the expected maternal inheritance was confirmed by PCR-RFLP (restriction fragment length polymorphism).     

基于线粒体COI基因分析钩手水母的群体遗传结构

钩手水母(Gonionemus vertens)为大西洋和太平洋广布种, 是我国习见的有毒水母种类之一。本文对采自黄渤海海域4个地理群体的104个钩手水母线粒体COI基因序列进行扩增, 并结合GenBank上其他182个钩手水母同源序列进行序列变异分析。在286个基因序列中共检测出52个多态位点, 定义了14种单倍型。总群体的单倍型多样性和核苷酸多样性分别为0.743 ± 0.012和1.046% ± 0.097%, 与其他几种大型水母相比, 钩手水母总群体的遗传多样性处于较高水平。AMOVA结果显示, 60.17%的分子变异源于群组间, 13.37%的分子变异源于群体内, 26.46%的分子变异源于组内群体间, 群组间、群体内和组内群体间的遗传分化均极显著。F_st值统计检验表明, 中国厦门群体与乐亭、东营、烟台、大连群体间存在显著的遗传分化, 大连与东营、烟台群体间也存在显著的遗传分化。系统分析结果显示, 钩手水母群体间存在2个明显的单倍型谱系分支。不同的钩手水母地理群体间具有复杂的遗传模式, 钩手水母复杂的生活史、扩散能力、地理隔离和海流分布可能是影响钩手水母遗传结构的重要因素。     

Genetic diversity of captive forest musk deer (Moschus berezovskii) inferred from the mitochondrial DNA control region

Forest musk deer ( Moschus berezovskii ) were once distributed widely in China. However, wild populations have declined dramatically because of poaching and habitat loss. Captive breeding populations have been established for several decades, but the genetic backgrounds of most captive populations were unclear and the population sizes increased very slowly. To provide useful information for conservation and management of this species, we investigated the genetic diversity and population structure of forest musk deer by analysing a 582-bp fragment of the mitochondrial DNA (mtDNA) control region (CR) in three captive breeding populations in Sichuan Province, China. Ninety-four variable sites and 27 haplotypes were observed in 109 individuals, and the nucleotide and haplotype diversities were relatively high compared with those of other endangered mammals. Of the three investigated populations, the Maerkang population had the highest nucleotide diversity ( π  = 0.0568), haplotype diversity ( h  =   0.836) and average intra-population genetic distance (0.062). The analysis of molecular variance demonstrated that most variation occurred within samples and that there was significant differentiation of the three populations. Estimates of gene flow indicated that there were few genetic exchanges among the three populations. Building pedigree records and increasing gene flow between populations will be helpful for conserving these populations and this species.     

Population structure in Indian sheep ascertained using microsatellite information

This study attempts to provide a comprehensive insight into the prevailing genetic status of Indian sheep breeds using microsatellite markers. Seventeen Indian sheep breeds from 3 agroecological zones were analysed using a panel of 25 microsatellite markers. All of the sheep breeds investigated were genetically diverse, as evident from the high allele (>6) and gene (>0.6) diversity values. The gene diversity values for all breeds ranged from 0.621 to 0.780. The within-population heterozygote deficit (F(IS)) varied from -0.098 to 0.234, reflecting significant levels for 12 of the 17 breeds investigated. The average genetic differentiation between all breeds (F(ST)) was 11.1%, revealing moderate discrimination between the indigenous sheep breeds. The genetic distance and principal component analysis revealed a separation of sheep breeds based on geographical propinquity. The Bayesian clustering approach suggested poor breed differentiation in the north-western arid and semi-arid region when compared to the breeds from the eastern and southern peninsular regions. The observed results mirror the divergent management strategies in the different agroecological regions, lack of specific selection policies, and intermixing of breeds in close proximity. Immediate steps to curb the intermixing and erosion of breed purity for some of these breeds need to be implemented, for example, by introducing measures like making proven rams available and ensuring their frequent exchange between flocks. The data generated here provides valuable information about the genetic structure of the 17 Indian sheep breeds and this can be used for designating priorities for their conservation.     

Genetic diversity and population structure of Schizopygopsis younghusbandi Regan in the Yarlung Tsangpo River inferred from mitochondrial DNA sequence analysis

The genetic diversity and population structure of Schizopygopsis younghusbandi from six sites in the middle reach of the Yarlung Tsangpo River were examined based on mitochondrial DNA cytochrome b (Cyt b) gene and control region (D-loop) sequences. 1141 bp of complete Cyt b sequences and 737 bp of partial D-loop sequences for 153 individuals of S. younghusbandi were obtained by using PCR amplification and sequencing. The results showed that S. younghusbandi populations had high haplotype diversity and low nucleotide diversity, and the population genetic diversity of this species was at a low level. Analysis of molecular variance revealed that most genetic variation occurred within populations, and that genetic differentiation was at low or moderate levels. The network of haplotypes based on Cyt b gene showed that there were two groups within the examined populations. Neutrality tests and mismatch distribution analysis suggested that this species might have undergone a population expansion and the expansion time was estimated to be 0.25–0.46 Ma BP. All these results would be crucial to establish scientific strategies for effective conservation and sustainable exploitation of wild resources of S. younghusbandi.     

Single genetic stock of kawakawa Euthynnus affinis (Cantor, 1849) along the Indian coast inferred from sequence analyses of mitochondrial DNA D-loop region

Kawakawa Euthynnus affinis is an epipelagic migratory tuna species, widely distributed in the tropical and subtropical waters of the Indo-Pacific region. Kawakawa constitutes the largest tuna fishery in the Indian waters. In the present study, genetic variation was assessed using sequence analyses of Mitochondrial DNA (mtDNA) D-loop region. A 500 bp segment of D-loop region was sequenced in 400 samples collected from eight localities (Veraval (VE), Ratnagiri (RA), Kochi (KO), Kavaratti (KA), Port-Blair (PB), Tuticorin (TU), Pondicherry (PO), and Vizag (VI)) along the Indian coast. Analysis of molecular variance of mtDNA data revealed no significant genetic differentiation among sites the (Φ _ST ?=?0.0028, P?=?0.20723) indicating a single population along the Indian coast. Phylogenetic analysis revealed no obvious phylogeographic pattern separating the eight samples of kawakawa. However, the genealogical relationships demonstrated that mtDNA D-loop sequences belong to two different clades (clade I and clade II). Clade I is the major clade which consists of more than 98?% specimens from each regional population while clade II has individuals from only three populations (VE, PO, and VI). Results of genetic analyses of the present study support a single stock management of kawakawa along the Indian coast.     

SSCP analysis of pig mitochondrial DNA D-loop region polymorphism

The sequence polymorphism that occurs in the mitochondrial DNA (mtDNA) displacement (D)-loop region is useful as a cytoplasmic DNA marker. We cloned the mtDNA D-loop regions of five breeds of pig by polymerase chain reaction (PCR) and determined their sequences. The sequence diversities in D-loop regions among five breeds of pig were located in the starting area of heavy-strand replication. From these sequences, we designed primers for PCR-mediated single-strand conformation polymorphism (PCR-SSCP) analysis that amplified the most polymorphic 227 bp fragment of the D-loop region. The results of PCR-SSCP analysis clearly showed that four types of polymorphism (A to D) are found in Landrace (A), Large White (A, B), Duroc (A), Göttingen miniature pig (B) and Meishan (C, D). The same polymorphisms were also detected from each porcine embryo by this method. Our results show that PCR-SSCP analysis is useful in detecting polymorphisms in the D-loop region of pigs and pig embryos.     

基于线粒体控制区的嘉陵裸裂尻鱼种群遗传结构分析

嘉陵裸裂尻鱼为青藏高原特有鱼类,近年来随自然地理气候的变迁和人类活动的影响,种群数量急剧减少。为了解嘉陵裸裂尻鱼的遗传背景以便更好的保护其遗传资源,本研究采用线粒体控制区部分序列变异,分析了嘉陵裸裂尻鱼6个地理种群的遗传结构和分布动态。在147尾个体中共发现17个变异位点,定义了14种单倍型,群体总的单倍型多样性较高为0.810,核苷酸多样性低为0.00698。AMOVA分析显示,44.29%的分子差异源于群体间,55.71%的分子差异源于群体内,群体间遗传分化极显著。Fst值统计检验表明,除宕昌群体和舟曲群体之间差异不显著外,其余两两群体之间Fst值统计检验均显著。基因流估计显示各群体间的基因流水平较高,遗传交流较频繁。Mantel test检验表明,嘉陵裸裂尻鱼种群之间遗传分化程度与地理距离存在显著相关。系统树和单倍型网络进化图显示,6个地理群体的单倍型按照嘉陵江水系和渭河水系形成两个大的类群。错配分布和中性检验表明嘉陵裸裂尻鱼群体在近期历史上群体大小保持稳定,未出现显著的种群扩张。根据本文所揭示的嘉陵裸裂尻鱼种群遗传结构特征,建议将分布在嘉陵江水系的嘉陵裸裂尻鱼作为一个整体进行保护,嘉陵裸裂尻鱼渭河种群属高度分化的单倍型类群,且遗传多样性极低,需对该种群进行优先保护。     

Genetic diversity of Asian water buffalo (Bubalus bubalis): mitochondrial DNA D-loop and cytochrome b sequence variation

Swamp and river buffalo mitochondrial DNA (mtDNA) was sequenced for 303 bp of the cytochrome b gene for 54 animals from 14 populations, and for 158 bp of the D-loop region for 80 animals from 11 populations. Only one cytochrome b haplotype was found in river buffalo. Of the four haplotypes identified in swamp buffalo, one found in all populations is apparently ancestral both to the other swamp haplotypes and to the river haplotype. The phylogenetic relationships among the 33 D-loop haplotypes, with a cluster of 11 found in swamp buffalo only, also support the evolution of domesticated swamp and river buffalo from an ancestral swamp-like animal, most likely represented today by the wild Asian buffalo ( Bubalus arnee ). The time of divergence of the swamp and river types, estimated from the D-loop data, is 28 000 to 87 000 years ago. We hypothesise that the species originated in mainland south-east Asia, and that it spread north to China and west to the Indian subcontinent, where the rive type evolved and was domesticated. Following domestication in China, the domesticated swamp buffalo spread through two separate routes, through Taiwan and the Philippines to the eastern islands of Borneo and Sulawesi, and south through mainland southeast Asia and then to the western islands of Indonesia.     

Large sequence divergence of mitochondrial DNA genotypes of the control region within populations of the african antelope, kob (Kobus kob)

The genetic differentiation among kob populations (Kobus kob) representing two recognized subspecies was examined using mitochondrial control region sequences. Two distinct lineages (estimated sequence divergence of 9.8%) exhibited different geographical distributions and do not coincide with previously recognized ranges of subspecies. The presence of the two lineages was further supported with sequences of mitochondrial cytochrome b gene. One lineage was predominant in the west and southern ranges of the populations studied and the other was commonly found in a more northern distribution (Murchison populations) in Uganda. Murchison and the geographically intermediate Toro populations (Uganda) represented the area of overlap. The existence of the two lineages in the area of overlap is hypothesized to have resulted from a range expansion and secondary contact of the two lineages of kob that evolved in allopatry. The existence of the kob during the Pleistocene offers a plausible explanation for the observed biogeographic pattern. Our mitochondrial data reveal two examples of discordance between a gene tree and presumed species tree as: (i) the two lineages co-occur in the kob subspecies, Kobus kob thomasi (Uganda kob); and (ii) the puku, which was included in the analysis because of its controversial taxonomic status (currently recognized as a distinct species from the kob), is paraphyletic with respect to the kob. Significant degrees of heterogeneity were detected between populations. Relatively high genetic variation was observed in the populations, however, the inclusion of distinct lineages influences the population structure and nucleotide diversity of the kob populations.     

黄淮海地区玉蜀黍黑粉菌群体遗传结构分析

利用17个多态性ISSR引物对2017年在我国黄淮海地区采集的41株玉蜀黍黑粉菌Ustilago maydis样本进行了种群遗传结构分析。基于UPGMA法和贝叶斯模型对玉蜀黍黑粉菌群体遗传结构分析显示,采集的样本可以分为两大群,且种群的划分表现出了与来源地理纬度的一致性。因此按照各样本所属地理纬度将样本分为北纬33°-34°、北纬35°-36°、北纬37°-38° 3大种群。结果表明,分离自北纬33°-34°地区的玉蜀黍黑粉菌遗传多样性最丰富,Nei’s基因多样性（H）为0.2216,Shannon指数（I）为0.3300。分子方差分析（AMOVA）显示,变异中来自种群内部的变异为86%,种群间变异14%（φpt=0.141）。     

Genetic diversity of golden takin (Budorcas taxicolor bedfordi) population from Qinling Mountains in China revealed by sequence analysis of mitochondrial DNA control region

The golden takin is an endangered species, listed as a First Grade Protected animal and found only in the Qinling Mountains in China. A great deal of research on the golden takin's living habitat, population size, and home range has been conducted. Here, we employed sequence analysis of the mitochondrial DNA control region to study the genetic diversity of the golden takin from three separate nature reserve parks in the Qinling Mountains. We also compared the results of our study with previously published data on the genetic diversity of mixed takin species located in the Qinling Mountains and the Minshan area. Based on 62 sampled golden takin individuals, we found an overall mean genetic haplotype diversity of 0.687. There is no significant geographic genetic diversity across different golden takin populations within the Qinling Mountains. However, we did show significant diversity between golden takin from the Qinling Mountains and from Minshan. These original data provide a foundation for the genetic diversity of golden takin, and will yield comprehensive information for better supporting the management in the national reserve parks.     

Population genetic structure and historical demography of grey mullet, Mugil cephalus, along the coast of China,inferred by analysis of the mitochondrial control region

Population genetic structure and historical demography of grey mullet, Mugil cephalus, along the coast of China were examined with a 389-bp segment of mtDNA control region. In total, 117 samples were collected from seven locations and 77 haplotypes were obtained. No haplotype was concurrently presented in all the samples, and samples in Southern and Northern regions had distinctly different haplotypes. Sorting of the data cladistically into phylogenetic trees indicated that these haplotypes can be divided into two main clades. Analysis of molecular variance (AMOVA) indicated significant genetic differentiation among subpopulations in different regions (φ_CT = 0.9505, p < 0.01). The average pairwise differences and φ_ST values (0.8943–0.9464) between regional populations were significant. The net Tumura and Nei genetic distance (0.5577 ± 0.1421) between the two groups was large. The deep and unique divergence between the two groups suggested that each group is likely to represent a distinct species. In contrast to the great genetic divergence on a broad scale, small but significant population subdivisions at a less spatial scale were also detected both in Southern and Northern regions, which support the conclusion that M. cephalus has limited dispersal potential. High haplotype diversity (h = 0.8716 ± 0.1462) and low nucleotide diversity (π = 0.0046 ± 0.0030) were found among the three Northern samples. Among the four Southern samples, however, extraordinarily high levels of both haplotype diversity (h = 0.9886 ± 0.0057) and nucleotide diversity (π = 0.0425 ± 0.0212) were detected. The different polymorphisms suggest an apparent distinguishing demographic history between regionally Southern and Northern populations, though mismatch distribution analyses and neutrality tests implied a late Pleistocene population expansion of both Northern and Southern populations.     

基于线粒体DNA序列的秦岭地区淡足青步甲种群遗传结构分析

淡足青步甲Chlaenius pallipes是一类重要的天敌昆虫。为了揭示秦岭地区淡足青步甲的种群遗传分化和种群历史动态,对13个地理种群151个个体的线粒体Cox1-tRNALeu-Cox2片段DNA序列进行了分析。序列比对后的全长为1 602 bp,共检测到57个多态性位点,定义65个单倍型,其中48个为居群内特有单倍型,17个为居群间共享单倍型。单倍型多样性指数H_d 为0.972,核苷酸多样性指数P_i为 0.0025。该种群遗传分化明显,但AMOVA分析结果表明变异主要来源于种群内,占变异总量的92.10%。SAMOVA和PERMUT分析结果均表明秦岭地区的淡足青步甲种群不存在明显的谱系地理结构。结合中性检验、错配分布和BSP分析结果表明该物种发生过种群扩张,且扩张时间大致在0.100-0.025 Ma之间。     

Genetic relationship of the melon fly,Bactrocera cucurbitae (Diptera: Tephritidae) inferred from mitochondrial DNA

Abstract The melon fruit fly, Bactrocera cucurbitae (Coquillett) (Diptera: Tephritidae), has been the subject of worldwide quarantine and management efforts due to its widespread agricultural impact and potential for rapid range expansion. From its presumed native distribution in India, this species has spread throughout the hot‐humid regions of the world. We provide information that reveals population structure, invasion history and population connectivity from 23 locations covering nine countries based on DNA sequences of the mitochondrial cytochrome oxidase I (COI) gene. Forty‐two polymorphic sites were described among 38 haplotypes. The most common haplotype, H1, was observed in 73% of the samples distributed among all populations. Highest genetic diversity was seen within populations, and no isolation‐by‐distance was detected. The western regions (Nepal, Bangladesh, Thailand, Burma and China‐west) showed higher haplotype diversity than eastern regions (China‐east). China‐Yunnan showed highest levels of genetic diversity in China. Haplotype diversity decreased with longitude from west to east. Together, these analyses suggest that B. cucurbitae has expanded from west to east within a limited geographic scale and recently invaded China through Yunnan Province.     

Population genetic structure, phylogeography and spawning philopatry in walleye (Stizostedion vitreum) from mitochondrial DNA control region sequences

Mitochondrial (mt) DNA control region sequences were used to test the genetic and phylogeographic structure of walleye Stizostedion vitreum populations at different geographical scales: among spawning sites, lake basins, lakes, and putative glacial refugia in the Great Lakes region. Sequencing 199 walleye revealed nucleotide substitutions and tandemly repeated sequences that varied in copy number, as well as in sequence composition, in 1200 bp of the mtDNA control region. Variable numbers of copies of an 11-bp tandem repeat showed no geographical patterning and were not used in further analyses. Substitutions in the other areas of the control region yielded 19 haplotypes, revealing phylogeographic structure and significant differences among glacial refugia, lakes, basins and some spawning sites. Differences among spawning populations were consistent with reduced gene flow, philopatry and possible natal homing. Analysis of spawning populations showed consistency of genotypic frequencies among years and between males and females, supporting philopatry in both sexes. The unglaciated plateau in southern Ohio, USA housed a very different haplotype that diverged prior to the Missouri, Mississippi and Atlantic glacial refugia types. Haplotypes from the three refugia colonized the Great Lakes after retreat of the Wisconsin glaciers, and their present distribution reflects the geography of their prior isolation and differential colonization. Populations that became associated with spawning localities appear to have diverged further due to philopatry, resulting in fine-scale phylogeographic structuring.     

褐马鸡圈养种群的mtDNA控制区多态性

褐马鸡(Crossoptilon mantchuricum)是我国特有的濒危鸟类,国家一级保护动物。为了保护褐马鸡的种质资源,从分子水平上评价褐马鸡的遗传多样性,对山西省庞泉沟自然保护区、太原市动物园的褐马鸡种群20个个体,线粒体DNA D-loop区全序列进行了克隆和测定,使用Clustal X、DnaSP4.0、Mega3.1等生物信息学软件,对全部20条序列开展了比对分析,确定了多态位点与单倍型数目,计算了核苷酸多样性和单倍型多样性;比较了两个种群的遗传变异,初步探讨了褐马鸡种群的遗传多样性和遗传结构。结果表明:20条褐马鸡线粒体DNA D-loop区全序列长度在1236 1237bp之间,其中A、T、G和C 4种核苷酸的平均含量分别为31.0%、26.8%、27.5%和14.8%,A+T含量(57.8%)高于G+C含量(42.3%)。排除1处核苷酸的插入或缺失后,共检测出26个突变位点,占分析序列总长度的2.1%,其中包括25处单一多态位点和1处简约信息位点。20个个体检测出13个单倍型,其中11个个体具有独特的单倍型,2个共享单倍型。褐马鸡两个种群的单倍型多样性(h)为0.911 0.933,核苷酸多样性(π)为0.002 0.003,单倍型间的遗传距离为0.003 0.002。根据单倍型构建了褐马鸡两个种群的NJ分子系统树。聚类表明,2个种群的个体并没有按相应的地理位置进行聚类。揭示褐马鸡具有较高的单倍型多样性和较低的核苷酸多样性,种群的遗传变异较低;两个种群单倍型间的遗传距离较近,遗传多样性参数接近,统计分析无显著差异,两个种群尚未表现出明显的遗传分化,且两个种群间有基因流存在。