Sequence polymorphism in a novel noncoding region of Pacific oyster mitochondrial DNA

Nucleotide sequence polymorphism in a 641-bp novel major noncoding region of mitochondrial DNA (mtDNA-NC) of the Pacific oyster Crassostrea gigas was analysed for 29 cultured individuals within the Goseong population. A total of 30 variable sites were detected, and the relative frequency of nucleotide alteration was determined to be 4.68. Alterations were mostly single nucleotide substitutions. Transition, transversion, both transition and transversion, and both transversion and nucleotide deletion were observed at 18, 9, 2 and 1 sites, respectively. Among 29 specimens, 22 haplotypes were identified, and pairwise genetic diversity of haplotypes was calculated to be 0.988 from multiple sequence substitutions using the two-parameter model. A phylogenetic tree, obtained for haplotypes by the neighbor-joining method, showed a single cluster of linkages. The cluster comprised 11 haplotypes associating with 14 specimens, while the other 11 haplotypes associating with 15 specimens were scattered. This mtDNA-NC presenting a high nucleotide sequence polymorphism is a potential mtDNA control region. It therefore can serve as a genetic marker for intraspecies phylogenetic analysis of the Pacific oyster and is more useful than the less polymorphic mtDNA coding genes.     

Genetic similarity of disjunct populations of the giant sea bass Stereolepis gigas

Sequence variation in nuclear and mitochondrial genes of the giant sea bass Stereolepis gigas collected from the Pacific coast and the northern Sea of Cortez was examined. Restriction fragment length polymorphism analysis and direct sequencing showed extremely low mtDNA sequence diversity (13 closely related haplotypes with no evidence of geographical population subdivision). The mitochondrial haplotype mismatch distribution is consistent with a population expansion following the Last Pleistocene glaciation. Differences in single nucleotide polymorphism frequencies between Pacific and Sea of Cortez populations were detected at two of four nuclear loci, which may reflect natural selection or genetic drift in populations with low effective numbers of males. Although Pacific coast and Sea of Cortez populations of giant sea bass do not exhibit the mitochondrial phylogenetic break characteristic of many species with disjunct Pacific and Gulf populations, the possibility of genetic differentiation at nuclear loci suggests that a cautious approach to broodstock selection for captive breeding and restoration programmes be exercised.     

PCR-RFLP genotyping for Japanese and Korean populations of Pacific oyster using mitochondrial DNA noncoding region

Production of Pacific oyster Crassostrea gigas in Miyagi and Hiroshima (Japan) has gradually increased, with a marked increase in imported oysters from Goseong (Korea), and then cultured oysters of the Miyagi, Hiroshima, and Goseong populations accounting for most oyster consumption in Japan. In this study, we developed a simple PCR-RFLP analysis of a mitochondrial DNA noncoding region (mtDNA-NCR) for differentiating cultured oysters of these populations. PCR amplification yielded a 818 bp fragment comprising the entire mtDNA-NCR and parts of adjacent tRNA^Cys and tRNA^Asn genes from all the specimens. By use of a wide restriction test using 30 different restriction enzymes, only a single enzyme only, AluI, produced a unique RFLP pattern enabling us to discriminate Miyagi and Hiroshima oysters from Goseong oysters. This difference is probably because of nucleotide alteration at the presumptive AluI recognizing site on position 439 of the mtDNA-NCR. Our simple, robust and cost-effective PCR-RFLP analysis is potentially useful for population genetic investigation of cultured Pacific oyster, particularly when large numbers of specimens must be analyzed.     

Genetic diversity and population structure of Sepiella japonica (Mollusca: Cephalopoda: Decapoda) inferred by mitochondrial DNA (COI) variations

In order to describe the genetic diversity of five geographical populations of cuttlefish (Sepiella japonica) along with Chinese coast and determined their phylogenetic relationship, partial mitochondrial COI gene (681bp in length) was amplified from 96 individuals collected from these populations and sequenced. The 5 populations of cuttlefish inhabit Yellow Sea, East China Sea and South China Sea. Out of 22 polymorphic nucleotides identified, 8 were represented by a single sequence, 12 were parsimony informative, which defined 22 haplotypes. Haplotype and nucleotide diversity were low among populations. Of 22 haplotypes, 15 appeared only in a single population, 6 appeared in 2 or 3 populations and 1 was shared by all populations. The COI gene was monomorphic in Qingdao population. The haplotypes identified clustered into 2 clades, each covered individuals from 5 populations each. Pairwise F_ST were not proportional to the geographical distances. Among the 5 populations, relatively high level of genetic diversity was found in Ningde population which was recommended to be the best choice of germplasm resources for artificial releasing project.     

Genetic Population Structure of Chum Salmon in the Pacific Rim Inferred from Mitochondrial DNA Sequence Variation

We examined the genetic population structure of chum salmon, Oncorhynchus keta, in the Pacific Rim using mitochondrial (mt) DNA analysis. Nucleotide sequence analysis of about 500 bp in the variable portion of the 5′ end of the mtDNA control region revealed 20 variable nucleotide sites, which defined 30 haplotypes of three genealogical clades (A, B, and C), in more than 2,100 individuals of 48 populations from Japan (16), Korea (1), Russia (10), and North America (21 from Alaska, British Columbia, and Washington). The observed haplotypes were mostly associated with geographic regions, in that clade A and C haplotypes characterized Asian populations and clade B haplotypes distinguished North American populations. The haplotype diversity was highest in the Japanese populations, suggesting a greater genetic variation in the populations of Japan than those of Russia and North America. The analysis of molecular variance and contingency χ² tests demonstrated strong structuring among the three geographic groups of populations and weak to moderate structuring within Japanese and North American populations. These results suggest that the observed geographic pattern might be influenced primarily by historic expansions or colonizations and secondarily by low or restricted gene flow between local groups within regions. In addition to the analysis of population structure, mtDNA data may be useful for constructing a baseline for stock identification of mixed populations of high seas chum salmon.     

Single-nucleotide polymorphisms and haplotype LD analysis of the 29-kb IGF2 region on chromosome 11p15.5 in the Korean population

OBJECTIVE: We investigated sequence variations of the 29-kb insulin-like growth factor 2 (IGF2) region in human chromosome region 11p15.5 in the Korean population. This region consists of IGF2, insulin-like growth factor 2 antisense (IGF2AS), and the insulin gene, all important candidate genes for various diseases, including cancer, obesity, diabetes, and coronary disease. While single nucleotide polymorphisms (SNPs) have been identified for this region and used in association studies, ethnic differences in genetic variation at this site have not been addressed. To date, SNPs for the entire 29-kb region in the Korean population have not been reported. METHODS: We surveyed a population of 108 Koreans for SNPs in the 29-kb IGF2 region. RESULTS: We identified 62 SNPs, consisting of 6 SNPs in the promoter region, 17 in the untranslated region, 19 in introns, and 20 in the intergenic region. We also analyzed linkage disequilibrium (LD) patterns and haplotypes using 36 high-frequency (> 5%)SNPs and found a well-defined LD block spanning about 13 kb that includes 8 kb of the IGF2AS gene, with two hot-spot regions flanking the LD block. CONCLUSION: These SNPs may be useful as genetic markers in disease association studies in the Korean population.     

Comparative analysis of the intergenic spacer regions and population structure of the species complex of the pathogenic yeast Cryptococcus neoformans

Cryptococcus neoformans is an opportunistic basidiomycete responsible for the high incidence of cryptococcosis in patients with AIDS and in other immune-compromised individuals. This study, which focused on the molecular structure and genetic variability of the two varieties in the C. neoformans and Cryptococcus gattii species complex, employed sequence analysis of the intergenic spacer regions, IGSI and IGSII. The IGS region is the most rapidly evolving region of the rDNA families. The IGSI displayed the most genetic variability represented by nucleotide base substitutions and the presence of long insertions/deletions (indels). In contrast, the IGSII region exhibited less heterogeneity and the indels were not as extensive as those displayed in the IGSI region. Both intergenic spacers contained short, interspersed repeat motifs, which can be related to length polymorphisms observed between sequences. Phylogenetic analysis undertaken in the IGSI, IGSII and IGSI +5S rRNA + IGSII regions revealed the presence of six major phylogenetic lineages, some of which segregated into subgroups. The major lineages are represented by genotypes 1 (C. neoformans var. grubii), genotype 2 (C. neoformans var. neoformans), and genotypes 3, 4, 5 and 6 represented by C. gattii. Genotype 6 is a newly described IGS genotypic group within the C. neoformans species complex. With the inclusion of IGS subgenotypic groups, our sequence analysis distinguished 12 different lineages. Sequencing of clones, which was performed to determine the presence of multiple alleles at the IGS locus in several hybrid strains, yielded a single IGS sequence type per isolate, thus suggesting that the selected group of cloned strains was mono-allelic at this locus. IGS sequence analyses proved to be a powerful technique for the delineation of the varieties of C. neoformans and C. gattii at genotypic and subgenotypic levels.     

Genetic diversity and population structure of wintering Western Sandpipers from the Sinaloa coast,Mexico

ABSTRACT Although Western Sandpipers (Calidris mauri) are one of the best‐studied shorebirds along the Pacific Flyway, their genetic identity and population structure is poorly known. We studied the genetic population structure of nonbreeding Western Sandpipers at Bahia Santa María in Sinaloa, northwestern Mexico. A 685‐bp fragment of the mtDNA control region was sequenced for 162 individuals, resulting in 29 variable positions that defined 41 haplotypes. The most common haplotype (WESA01) occurred in 44% of all individuals, whereas 15 were unique to single individuals. Nucleotide diversity was low (π= 0.0030 ± 0.0019 [SE]), but haplotype diversity was moderately high (h= 0.802 ± 0.033 [SE]). The main two maternal lineages exhibited a slightly different local scale distribution that appeared to be related to migratory chronology. Lineage A represented 71% of the haplotypes and was evenly distributed across the nonbreeding season and habitat types, whereas Lineage B represented only 29% of the haplotypes and was disproportionately represented in January in some habitat types. Overall, the low level of nucleotide diversity, the star‐shape of the haplotype tree, the mismatch distribution, and the significantly negative Tajima's D values suggest that Western Sandpipers underwent a recent demographic expansion. Although our results are based on a small sample size from one of several wintering sites along the Pacific Flyway, Santa María likely contains a relatively high proportion of the species genetic variability because it is the wintering ground of nearly 10% of the global population.     

Population Genetic Structure of the Ark Shell <Emphasis Type="Italic">Scapharca broughtonii</Emphasis> Schrenck from Korea,China, and Russia Based on <Emphasis Type="Italic">COI</Emphasis> Gene Sequences

Haplotype distribution, gene flow, and population genetic structure of the ark shell (Scapharca broughtonii) were studied using a partial sequence of a mitochondrial COI gene. The sequence analysis of 100 specimens obtained from a total of seven localities-five in Korea, one in China, and one in Russia- revealed 29 haplotypes, ranging in sequence divergence from 0.1% to 2.1%. Among these, the most frequent haplotype, SB16, was extensively distributed over study areas, especially in all Korean localities. This extensive distribution consequently resulted in the near absence of statistically significant genetic distance. Also, a high rate of gene flow was characteristic among localities in Korea. A test of genetic population structure showed that the ark shell in Korea formed a large genetic group. Moreover, an AMOVA test to determine the allocation of the genetic variance showed that most of the variance was distributed between localities, instead of within localities. However, a significant population differentiation was found between geographic populations [i.e., Jinhae (locality 6) in Korea and Sangdong (locality 5) in China and Vladivostok (locality 7) in Russia] based on geographic distance and population structure. These distinct groups may be associated with geographic characteristics and barriers. The results suggest that most of the ark shell populations in Korea caused considerable distribution to form a genetically homogeneous and intermixing structure, whereas some of the Korean and Chinese and Russian populations had a significantly different genetic structure.     

铜鱼线粒体控制区的序列变异和遗传多样性

采用PCR和DNA测序技术研究长江中上游野生铜鱼的遗传多样性和群体遗传学特征,从9个采样点共获得100尾铜鱼,用于分析的线粒体DNA控制区的片段序列为946bp。在100个序列中,共检测出变异位点47个(其中增添/缺失位点8个),单倍型41种。9个地理群体的平均单倍型多样性(Hd)和平均核苷酸多样性(Pi)分别为0.9257±0.0162和0.004178±0.002337,表现出较贫乏的遗传多样性。群体间的分化指数(FST值)、平均基因流(Nm)、分子方差分析(AMOVA)和平均K2-P遗传距离均表明9个铜鱼地理群体间存在广泛的基因交流,未明显发生群体遗传分化。另外,共享单倍型比例较高,约为34%(14/41)。单倍型的UPGMA分子系统树和简约网络图显示单倍型的聚类与地理分群没有相关性。上述结果表明9个铜鱼地理群体属于同一种群。       

西藏牦牛线粒体DNA 的遗传多样性及系统进化分析

为从分子水平上探究西藏牦牛类群的遗传多样性、亲缘关系,本研究测定了日多牦牛、类乌齐牦牛、丁青牦牛、错那牦牛、隆子牦牛、仲巴牦牛、聂荣牦牛、申札牦牛等8 个西藏牦牛类群共328 头牦牛mtDNAD-loop区序列,分析其多态性,构建系统进化树。结果表明:本次测定的西藏牦牛mtDNA D-loop 区序列长度为 887 - 895 bp,共检测到135 个变异位点,其中单态突变位点52 个,简约信息位点83 个。在328 个个体中共检测出91 种单倍型,平均单倍型多样性(Hd)、平均核苷酸多样性(π)分别为0. 884、0.010 27,显示西藏牦牛具有丰富的遗传多样性。8 个类群间平均遗传距离为0.011;日多牦牛与错那牦牛间遗传距离最小(0. 006);类乌 齐牦牛与隆子牦牛间遗传距离最大(0.015)。系统进化分析显示西藏牦牛可分为两大类,错那牦牛是较纯的牦牛类群,其它牦牛类群在进化过程中出现相互交流的情况。     

PCR‐RFLP markers detect 29 mitochondrial haplotypes in Pacific lamprey (Entosphenus tridentatus)

We developed five polymerase chain reaction‐based markers that detect variation in the mitochondrial genome of the Pacific lamprey, Entosphenus tridentatus, across most of its range. Two gene fragments (ND2 and ND5) were amplified and digested with three and two restriction enzymes, respectively, detecting sequence variation at 18 sites (ND2 = 13; ND5 = 5) and yielding 29 composite haplotypes among 1246 lampreys. These sequence‐based markers will be useful in a range of phylogeographical and population genetic studies.     

小口白甲鱼都柳江种群mtDNA D环的序列变异及遗传多样性

采用PCR结合DNA测序技术,测定分析了易危鱼类小口白甲鱼(Onychostoma lini)都柳江种群36个个体mtDNA D环约470bp序列的变异及遗传多样性。结果表明,在36个个体中,该序列的长度为469～475bp,其碱基组成为A+T的平均含量(68.4%)高于G+C(31.6%)。共检测到25个多态位点,其中转换19个、颠换6个。核苷酸多样性(π)为0.00575,平均核苷酸差异数(K)为2.695。36个个体分属5个单倍型,单倍型多样度(Hd)为0.260,单倍型间的平均遗传距离(P)为0.026。5个单倍型构建的UPGMA系统树聚为2个分支。目前小口白甲鱼都柳江种群mtDNA D环序列存在着较丰富的变异和遗传多样性。     

The marine indo-west pacific break: contrasting the resolving power of mitochondrial and nuclear genes

Simultaneous studies of both nuclear and mitochondrial markerswere undertaken in two widespread Indo-West Pacific (IWP) marineinvertebrates to compare and contrast the ability of these markersto resolve genetic structure. In particular, we were interestedin the resolution of a genetic break between the Indian andPacific Oceans due to historical isolation. Sequence variationfrom the nuclear gene encoding myosin heavy chain (MyHC) andthe mitochondrial gene cytochrome oxidase I (COI) were examinedfor the snapping shrimp Alpheus lottini from wide-ranging populationsthroughout the Indian and Pacific Oceans. A previously identifiedgenetic break between oceans based on COI sequences appearsto have been an artifact caused by the inadvertent inclusionof pseudogene sequences; our new COI data provide evidence onlyof a break between IWP and East Pacific populations. Distributionof a single nucleotide polymorphism in MyHC, on the other hand,shows evidence of a cline between Indian and Pacific Oceans.New allozyme and mtDNA sequence data were also obtained forthe starfish Linckia laevigata. Allozyme data show a clear geneticbreak between Indian Ocean populations and Pacific (includingwestern Australian) populations, whereas the distribution ofmtDNA haplotypes shows a region of overlap in the central IWP.Comparisons of our data for both Alpheus and Linckia with datafrom other population genetic studies in the IWP suggest thatnuclear markers (allozymes, sequence data and morphologicalcharacters) may in some instances reveal historical patternsof genetic population structure whereas mtDNA variation betterreflects present day patterns of gene flow.     

Discovery of multiple IGS haplotypes within genotypes of Puccinia striiformis

In a search for specific molecular markers for population analysis of Puccinia striiformis f. sp. tritici, the ribosomal DNA (rDNA) intergenic spacer (IGS) 1 region (rDNA-IGS1, between the 28S and the 5S rDNA genes) was amplified, cloned, and sequenced. It was found to exhibit multiple bands and length polymorphism. Surprisingly, single isolates were found to possess between three to five different IGS1 haplotypes. Bands were cloned and sequenced, and two highly variable regions (α and β) were found between conserved regions, with repeat units interspersed in both types of regions. There were 14 different repeat units, and these were sometimes grouped further into four combinations of repeat units, with a few individual nucleotides (A or C) inserted between the repeats. Among three geographically dispersed isolates, the variable region α was divided into eight types, and the variable region β was divided into two types based on repeat units. Most of the 14 repeat units were shared by the variable and the conserved regions. Among the three isolates, there were a total of 12 IGS1 haplotypes, but some of these were shared between isolates such that there were only eight unique haplotypes. The occurrence of multiple haplotypes within single isolates may be useful for analyzing the population structure, tracking the origin of annual epidemics and providing insights into evolutionary biology of this pathogen.     

Phylogeographical analysis of an estuarine fish, Salanx ariakensis (Osmeridae: Salanginae) in the north-western Pacific

This study extended the geographic coverage of a previous study to explore population genetic structure and demographic history in the Ariake icefish Salanx ariakensis from three populations of continental coastlines and one island population in the north-western Pacific based on a partial sequence of the mitochondrial cytochrome b gene. The S. ariakensis showed high genetic diversity and strong genetic structure. Phylogenetic analysis showed a shallow gene tree with no clear phylogeographical structure. Contiguous range expansion and restricted gene flow were inferred to be main population events by nested-clade analysis. Significant genetic differentiations between populations could be attributable to negligible gene flow by coalescent analysis. High nucleotide diversity of each population was due to geographic mixing of heterogenous haplotypes during lowering sea levels of the Pleistocene. These findings indicate that cycles of geographic isolation and secondary contact happened in the Pleistocene glacial–interglacial cycles shaping genetic structure and population demography of S. ariakensis .     

缅甸蟒线粒体Cytb基因遗传多样性分析

为了探讨海南文昌种蟒养殖基地的种蟒的遗传多样性和个体来源,本研究对27条种蟒的细胞色素b基因的全序列进行了测定和分析。结果表明:缅甸蟒cytb基因全长1 111 bp,序列中A+T含量为58.0%,G+C含量为42.0%,单倍型多样性(Hd)为(0.755±0.053),核苷酸多样性(Pi)为(0.003 29±0.000 43),表明该种群遗传多样性较高。共检测出6条单倍型,单倍型间遗传距离最大为0.007,最小为0.001;与已知地理来源的个体构建系统发育树显示,该养殖种群来源两个母系世系。本研究结果为制定缅甸蟒合理圈养计划和科学放归提供理论基础。     

Trans‐Pacific and trans‐Arctic pathways of the intertidal macroalga Fucus distichus L. reveal multiple glacial refugia and colonizations from the North Pacific to the North Atlantic

Aim We examined the phylogeography of the cold‐temperate macroalgal species Fucus distichus L., a key foundation species in rocky intertidal shores and the only Fucus species to occur naturally in both the North Pacific and the North Atlantic. Location North Pacific and North Atlantic oceans (42° to 77° N). Methods We genotyped individuals from 23 populations for a mitochondrial DNA (mtDNA) intergenic spacer (IGS) (n = 608) and the cytochrome c oxidase subunit I (COI) region (n = 276), as well as for six nuclear microsatellite loci (n = 592). Phylogeographic structure and connectivity were assessed using population genetic and phylogenetic network analyses. Results IGS mtDNA haplotype diversity was highest in the North Pacific, and divergence between Pacific haplotypes was much older than that of the single cluster of Atlantic haplotypes. Two ancestral Pacific IGS/COI clusters led to a widespread Atlantic cluster. High mtDNA and microsatellite diversities were observed in Prince William Sound, Alaska, 11 years after severe disturbance by the 1989 Exxon Valdez oil spill. Main conclusions At least two colonizations occurred from the older North Pacific populations to the North Atlantic between the opening of the Bering Strait and the onset of the Last Glacial Maximum. One colonization event was from the Japanese Archipelago/eastern Aleutians, and a second was from the Alaskan mainland around the Gulf of Alaska. Japanese populations probably arose from a single recolonization event from the eastern Aleutian Islands before the North Pacific–North Atlantic colonization. In the North Atlantic, the Last Glacial Maximum forced the species into at least two known glacial refugia: the Nova Scotia/Newfoundland (Canada) region and Andøya (northern Norway). The presence of two private haplotypes in the central Atlantic suggests the possibility of colonization from other refugia that are now too warm to support F. distichus. With the continuing decline in Arctic ice cover as a result of global climate change, renewed contact between North Pacific and North Atlantic populations of Fucus species is expected.     

Genetic similarity between the South Atlantic and the western North Pacific Maurolicus (Stomiiformes: Actinopterygii) taxa,M. walvisensis Parin & Kobyliansky and M. japonicus Ishikawa: evidence for synonymy?

An analysis of the 524 nucleotide long segment in the 16SrDNA mitochondrial gene of Maurolicus japonicus revealed that there are two major haplotypes (E and H) and two minor haplotypes (I and J), which comprise 63·6, 27·2, 4·6 and 4·6% of the population, respectively. The nucleotide sequence of the major haplotype E is identical to that of the most common haplotype in Maurolicus walvisensis (63·6% of the population). The other haplotypes of M. japonicus are almost identical to that of the haplotype E with only a single (haplotypes I and J) or three nucleotide differences (haplotype H). Phylogenetic trees of all the 16SrDNA haplotypes found thus far in the Maurolicus taxa show that the relationships among the haplotypes of M. japonicus and M. walvisensis are indistinguishable but that they are clearly distinctive from those of Maurolicus muelleri . Examination of the morphometric characteristics of specimens reveals similarities among the individuals of different haplotypes of M. japonicus and also between M. japonicus and M. walvisensis in almost all characteristics. The results suggest that despite the current ocean-wide allopatric distribution between M. japonicus and M. walvisensis , the two taxa are conspecific as M. japonicus .     

基于线粒体Cyt b基因的龙头鱼群体遗传结构分析

在我国黄海、东海和南海海岸带上, 选取青岛、南通、舟山、三门、宁德、泉州和湛江共7个群体164尾龙头鱼(Harpadon nehereus)为研究对象, 通过PCR扩增共获得长度为1112 bp的线粒体DNA细胞色素b (Cyt b)基因序列, 共检测到32个变异位点, 其中单变异位点27个, 简约信息位点1个。164条序列定义了29个单倍型, 平均单倍型多样性(H_d)和核苷酸多样性(π)分别为0.3026±0.0479和0.000371±0.000379, 其中泉州群体的单倍型多样性和核苷酸多样性均最低。分析比较了不同龙头鱼群体间遗传变异情况, 发现群体间平均遗传距离为0.00035, 遗传分化指数F_ST值均小于0.05, 群体间没有发生明显的遗传分化。AMOVA分析结果显示龙头鱼群体遗传差异主要来源于群体内个体间的变异。中性检验的Tajima’s D和Fu’s F_s统计值均为负值且显著偏离中性, 核苷酸不配对分布图呈现明显的单峰分布, 表明龙头鱼历史上经历了群体扩张事件, 参考已知海洋鱼类Cyt b基因2%每百万年的进化速率, 估算群体扩张发生的时间大约在0.08—0.32百万年前的第四纪更新世中晚期。