Genetic diversity of wild mekong giant catfish Pangasianodon gigas collected from Thailand and Cambodia

The Mekong giant catfish Pangasianodon gigas is endemic to the Mekong River and is a critically endangered species. The genotypes of the microsatellite DNA (msDNA) and mitochondrial DNA (mtDNA) markers (right domain of the control region) were detected to evaluate the present status of genetic divergence of this species from the Mekong River in Thailand and Cambodia. The observed and expected heterozygosity values of Mekong giant catfish in Thailand and Cambodia were relatively low in comparison with those of other nonendangered freshwater fish species. These two populations from Thailand and Cambodia showed similar levels of genetic diversity, as evaluated by the 384 nucleotides of the mtDNA control region with 13 haplotypes. The pairwise F _ST value between the two populations based on the genotype frequencies of msDNA and mtDNA markers suggested a close genetic relationship between the populations in Thailand and Cambodia. The results of this study support the conclusion that the Mekong giant catfish is critically endangered. Care should be taken to sustain the genetic diversity of this species, as the level of genetic variability has already decreased in the wild population. This species is a target species for an ongoing stock enhancement program in the Mekong River in Thailand. It is proposed to apply these markers for proper broodstock management, such as for minimal kinship selective breeding in the hatchery.     

Microsatellite markers reveal population structure in gilthead sea bream Sparus auratus from the Atlantic Ocean and Mediterranean Sea

ABSTRACT:   This study was carried out to investigate the genetic variability and the population genetics of Sparus auratus. In fact, despite its importance in Mediterranean fisheries and aquaculture, very little it is known concerning its population structure. Samples of wild gilthead sea bream were collected in seven different localities along the Mediterranean and Atlantic coasts and were genetically characterized by means of microsatellite markers. Genotyping at four microsatellite loci revealed high polymorphism (7–38 alleles/locus) and expected heterozygosities, which ranged from 0.80 to 0.85. A slight but significant population structure was found ( F _ST = 0.010). In fact, at least three populations of gilthead sea bream within the Western Mediterranean Sea were identified (Sardinian Sea, Sardinian Channel and Central Tyrrhenian Sea), which are also genetically differentiated from those of the Atlantic Ocean and the Adriatic Sea.     

Genetic structure of the migratory catfish Pseudoplatystoma corruscans (Siluriformes: Pimelodidae) suggests homing behaviour

Abstract –  Pseudoplatystoma corruscans is distributed widely in the La Plata Basin in Brazil, where it is a very important species because of its high commercial value and ecological role as a voracious predator. This species undertakes long migrations during the dry and rainy seasons to feed and reproduce, respectively. In this study, we analysed seven microsatellite loci to test the hypotheses of the existence of a single panmictic population of P. corruscans in the La Plata Basin. All microsatellites analysed were highly polymorphic with the number of alleles per locus ranging from seven (Pcor28) to 30 (Pcor10). Across all samples, 107 alleles were detected. Thirty alleles were private, i.e., found in only one sample. Highly significant genetic differentiation was observed among samples in three of the four analyses performed: F _ST (from 0.03435 to 0.16349, P  < 0.05), Analysis of molecular variance (amova) F _ST = 8.25% ( P  < 0.0001) and amova R _ST = 9.72% ( P  < 0.0001). A microsatellite multilocus Bayesian assignment test with the programme structure confirmed the division of the fishes into six groups largely concurrent with main branches on a population neighbour-joining tree. The obtained results rejected the panmixia hypothesis. On the other hand, the documented long-distance movements of P. corruscans and the geographical scale of genetic differentiation found in this study indicate a strong tendency for fish of this species to utilise their natal nursery regions for reproduction, which represents the first demonstrated example of homing in a freshwater catfish in South America.     

Current status of freshwater prawn culture in Vietnam and the development and transfer of seed production technology

ABSTRACT:   In Vietnam, the giant freshwater prawn Macrobrachium rosenbergii is becoming an increasingly important targeted species, as its culture, especially in rice fields, is considered to have the potential to raise income among impoverished farmers. The production of M. rosenbergii based on aquaculture reached over 10 000 tons per year in 2002, having increased from about 2500 tons since the 1990s. Until recently, lack of a stable supply of seed had been an important obstacle to the further expansion and development of M. rosenbergii culture, but cumulative research on larval rearing, especially in the 1990s, has led to the development of new seed production technology based on the 'modified stagnant green water system'. Following its dissemination by the efforts of provincial authorities, hatchery operators, and farmers, the freshwater prawn seed production industry developed rapidly in the Mekong Delta with over 90 hatcheries producing 76.5 million postlarvae in 2003. This is considered to have affected the expansion of rice–prawn farming in the Mekong Delta, leading to increased aquacultural production in the region. This paper reviews the current status of freshwater prawn culture in Vietnam and background history, and presents a socioeconomic evaluation of seed production technology implementation.     

Population structure and partial anadromy in Oncorhynchus mykiss from Kamchatka: relevance for conservation strategies around the Pacific Rim

Abstract –  Conservation of life-history diversity found in Oncorhynchus mykiss requires knowledge of the underlying population structure and genetic basis of this variability. We analysed variation at 10 microsatellite loci from seven rivers across Kamchatka to identify population structure and to test for divergence between life-history forms. We found lower heterozygosity in Kamchatkan populations compared with North American populations, but population structure was substantial (region-wide F _ST = 0.11) and followed an isolation-by-distance pattern similar to that reported for older North American populations. We found no evidence for genetic divergence between resident and anadromous individuals in the Sopochnaya River or between typically anadromous individuals and 'half-pounders' in the Utkholok River. A review of other studies of reproductive isolation, in combination with our results, suggests: (1) that pristine populations of steelhead should be expected to exhibit partial anadromy; and (2) that managing anadromous and resident individuals separately without demonstrating reproductive isolation is biologically unsound.     

五个斑点叉尾群体的微卫星遗传多样性分析

用8对微卫星引物对1997-2004年引进的5个斑点叉尾群体进行遗传多样性分析,计算并统计等位基因数、多态信息含量（PIC）、杂合度、遗传相似性系数、遗传距离等参数。实验显示8个微卫星位点在5个斑点叉尾群体中共检测到42个等位基因,平均期望杂合度为0.6338～0.7320,表明其遗传多样性程度处于中等偏上水平。平均多态信息含量为0.5756～0.6869,说明基因座为高度多态基因座（PIC〉0.5）。群体间遗传相似系数为0.7504～0.9203。聚类分析显示,04群体与其他4个群体的亲缘关系较远。结果表明：引进的5个斑点叉尾群体均具有较高的遗传多样性,遗传信息丰富,遗传变异大,为良好的育种材料。     

西南大西洋阿根廷滑柔鱼遗传多样性的微卫星分析

西南大西洋阿根廷滑柔鱼(Illex argentinus)是重要的经济大洋性头足类,其种群结构较为复杂。本研究采用8个微卫星标记,对西南大西洋阿根廷滑柔鱼群体的遗传多样性进行了检测。8个基因座位上,共检测出172个等位基因,每个基因座位检测到10～30个等位基因不等。8个位点的平均观测等位基因数为21.500 0,平均有效等位基因数为12.074 1。各位点的观测杂合度(Ho)变化范围为0.300 0～0.775 0,期望杂合度(He)为0.484 2～0.977 0,平均观测杂合度为0.504 6,平均期望杂合度为0.873 4。8个微卫星位点的多态信息含量(PIC)平均值为0.853 6。以卡方检验估计Hardy-Weinberg平衡,发现8个基因座位均不符合Hardy-Weinberg平衡。Fst分析显示该群体无明显的遗传分化。研究结果揭示:西南大西洋阿根廷滑柔鱼种群显示了高水平的微卫星多态性,该海域阿根廷滑柔鱼群体的遗传多态性水平在经历了近十年的密集捕捞压力后没有明显的变化,同时证明微卫星分子标记是研究该物种自然种群的有利工具。     

Genetic divergence in Indian populations of M. rosenbergii using microsatellite markers

Macrobrachium rosenbergii, known as the giant freshwater prawn or Malaysian prawn, is the sixth largest aquaculture species in Asia. Knowledge of genetic diversity of M. rosenbergii is important to support management and conservation programmes, which will subsequently help in sustainable production of this economically important species. This study aimed to analyse the genetic diversity and population structure of five M. rosenbergii populations using 11 microsatellite loci. In analysing 240 samples, the number of alleles, observed heterozygosity (H_O) and expected heterozygosity (H_E) ranged from 3 to 20, from 0.250 to 0.978 and from 0.556 to 0.944 respectively. The five stocks of M. rosenbergii displayed high level of genetic diversity. Both the F_ST and amova analyses showed that there was significant genetic differentiation among all populations. The UPGMA dendrogram based on Nei's genetic distance matrix revealed that the Narmada and Mahi populations were in one cluster and Mahanadi and Subarnarekha populations in another single major branch, whereas the Kerala population clearly showed a separate cluster. This information on genetic variation will be useful for genetic improvement and conservation of Indian populations of giant freshwater prawn M. rosenbergii.     

Bioaccumulation and distribution of trace elements in tissues of giant river prawn <Emphasis Type="Italic">Macrobrachium rosenbergii</Emphasis> (Decapoda: Palaemonidae) from South Vietnam

ABSTRACT:   Accumulation profiles of 22 trace elements in abdominal muscle, abdominal exoskeleton and the hepatopancreas of the giant river prawn Macrobrachium rosenbergii were analyzed. The giant river prawn is an indigenous freshwater species from South Vietnam, and is cultured commercially and fished in the wild. Samples were collected from Ho Chi Minh City and the surrounding area (SKEZ, South Key Economic Zone), and from the Mekong River Delta between 2003 and 2005. Highest accumulations of essential (Cu, Se and Mo) and toxic (As, Ag, Cd and Hg) elements were observed in hepatopancreatic tissue, except for Mn, Sr, Sn, Ba and V in the exoskeleton and Rb and Cs in muscle tissue. Spatial differences showed concentrations of Cs and Pb in muscle and Sr in exoskeletons from the SKEZ were higher than those from the Mekong River Delta. The opposite trend was observed for Cr, Se and Sb in muscle, Mo, Sb and Tl in exoskeleton, and Se, Hg, Mo, Cd, Sb, Tl and Bi in the hepatopancreas. These differences in trace element concentrations in prawns likely reflect differences in industrialization and human activities between the two regions of South Vietnam.     

Genetic diversity and population history of the migratory catfishes Pangasianodon hypophthalmus and Pangasius bocourti in the Cambodian Mekong River

ABSTRACT:   Polymerase chain reaction (PCR) restriction fragment length polymorphism analysis of mitochondrial DNA was applied to the genetic structure and evolutionary history of the more ancestral Pangasianodon hypophthalmus ( n  = 82), and the recently speciated catfish Pangasius bocourti ( n  = 90) from the Cambodian Mekong River. Both pangasiids were characterized by a lack of genetic population structure that may result from high levels of contemporary gene flow. Genetic diversity was lower in P. hypophthalmus than in P. bocourti . However, a different evolutionary history was inferred for both species based on genealogical and demographic analyses (mismatch analysis, Tajima's D- and Fu's F _S-tests). The genetic profile of the more ancestral P. hypophthalmus shows indications of a recent population bottleneck, whereas the recently speciated P. bocourti shows signatures of historical population expansion. This study stresses the importance of preserving the migration routes in the Cambodian Mekong basin in order to maintain the genetic diversity and long-term integrity of both species.     

Genetic difference between Ezo-awabi <Emphasis Type="Italic">Haliotis discus hannai</Emphasis> and Kuro-awabi <Emphasis Type="Italic">H. discus discus</Emphasis> populations: Microsatellite-based population analysis in Japanese abalone

ABSTRACT:   This study documents the genetic population relationships between Ezo-awabi Haliotis discus hannai and Kuro-awabi H. discus discus by means of the microsatellite technique with an emphasis on the extent of genetic difference between Ezo- and Kuro-awabi populations. Eight markers were employed to screen five populations each collected from Ezo- and Kuro-awabi habitats. All eight loci showed polymorphisms in all populations (number of alleles per locus, 11.1–14.1; averaged expected heterozygosity, 0.64–0.70). The number of loci that yielded significant genetic heterogeneities (allele frequency distribution and/or pairwise θ ) between Ezo- and Kuro-awabi populations was larger than that generated in comparisons between populations within Ezo- and Kuro-awabi habitats. According to the bootstrap neighbor-joining trees constructed on the basis of two genetic distance measures ( D _A and D _ST), the 10 populations fell into two clusters of populations (Ezo-awabi and Kuro-awabi habitat groups), and the results of hierarchical amova testing also supported the neighbor-joining clustering. The outcomes presented here suggest that the microsatellite markers used in this study are potentially an efficient means to differentiate Ezo- and Kuro-awabi populations.     

Genetic differentiation between Zhe oyster Crassostrea plicatula and pacific oyster Crassostrea gigas populations in China assessed by microsatellite analysis

ABSTRACT:   Genetic differentiation and relationships between Crassostrea plicatula and Crassostrea gigas populations from China were studied by means of the microsatellite technique. Seven loci were used to screen five populations each collected from C. plicatula and C. gigas . All loci showed high polymorphism for all populations, as observed in average number of alleles per locus (19.1–28.1), and average expected heterozygosity (0.891–0.954). Significant departures from Hardy–Weinberg equilibrium due to heterozygote deficiency were observed over most populations at each locus and were best explained by null alleles. F _ST values showed significant genetic differentiation between C. plicatula and C. gigas populations. According to the neighbor-joining tree constructed on the basis of the genetic distance ( D _A), the ten populations fell into two distinct groups ( C. plicatula and C. gigas groups), and the results of principal coordinate analysis and assignment tests also supported the neighbor-joining clustering. The outcomes presented here suggested that the microsatellite markers have great potential for differentiating C. plicatula from C. gigas populations. The information obtained in this study has important implications for the suitable management and conservation of these genetic resources in China.     

Evidence for strong genetic structure in a regionally important, highly vagile cyprinid (Henicorhynchus lobatus) in the Mekong River Basin

Abstract –  For effective management of a freshwater species, it is necessary to have an understanding of the spatial scale of population differentiation. In many cases, common ecological methods such as mark-recapture are inappropriate for large freshwater systems. In this study, a molecular genetics approach was employed to determine the population structure of a small, but regionally very important, cyprinid species ( Henicorhynchus lobatus ) in the lower Mekong River Basin (MRB). A fragment of the mtDNA ATP ase 6 and 8 genes was sequenced for 136 individuals sampled from the main Mekong River channel, from two major tributaries of the Mekong River and from a site in an adjacent river drainage (Chao Phraya). An exact test and samova analysis revealed considerable differentiation among sites from the Mun River (a tributary of the Mekong River in Thailand) and all other samples from the MRB. Furthermore, the most upstream site in Lao PDR (BP) was significantly differentiated from sites further downstream. It is argued here that the former pattern is a result of historical drainage rearrangement and the latter a function of current ecological differences expressed as discrete migration pathways. The results of the study clearly show the existence of discrete population units for H. lobatus within the MRB that may require separate management strategies.     

黄颡鱼(♀)、长吻[鱼危](♂)及其杂交F1代遗传多样性分析

利用SSR分子标记技术和从20对微卫星引物中筛选出的12对多态性较高的引物,分析1龄、体质量(14.20±1.20)g黄颡鱼(♀)和长吻[鱼危](♂)及其杂交F1代[黄颡鱼(♀)×野生长吻[鱼危](♂)]的遗传多样性。试验结果显示,这3个群体之间的平均等位基因数分别为4.583、3.667、5.000;平均有效等位基因数分别为2.382、2.221和2.535;平均观测杂合度分别为0.419、0.367和0.604;平均期望杂合度分别为0.477、0.383和0.597;多态信息含量分别为0.420、0.365和0.509。杂交F1代与父本长吻[鱼危]的遗传距离为0.8551,小于与母本黄颡鱼的遗传距离(1.7271),在亲缘关系树状图上先与父本聚为一支。杂交F1代从父本中获得较多的遗传物质,较双亲有丰富的遗传多样性和更高的基因杂合度。该试验结果将为研究黄颡鱼和长吻[鱼危]杂交后代的遗传多样性,探索鲿科鱼类属间杂交育种后亲本与子代的亲缘关系和遗传规律提供理论依据。     

三角帆蚌微卫星位点筛选及多态性分析

采用磁珠富集法,以生物素标记的(CA)10寡核苷酸为探针,构建了三角帆蚌(Hyriopsis cumingii)基因组微卫星富集文库。根据微卫星位点的侧翼序列设计引物,随机挑选扩增出与预期大小相符的32对引物,引物荧光标记后对24个三角帆蚌个体分别进行PCR扩增,共筛选出20对多态性较好的引物。结果表明,20个微卫星位点的等位基因数为5～20,有效等位基因数2.321 3～13.260 3。观察杂合度和期望杂合度分别为0.318 2～1.000 0和0.582 5～0.946 1;PIC值0.547 2～0.919 9;其中14个位点符合Hardy-Weinberg平衡(P>0.05)。本研究筛选的20个微卫星标记可作为三角帆蚌遗传多样性、种群遗传结构等研究的理想分子标记。     

Genetic differentiation of the bronze featherback Notopterus notopterus between Mekong River and Tonle Sap Lake populations by mitochondrial DNA analysis

ABSTRACT:   Although the bronze featherback Notopterus notopterus is an important fish in South-East Asia, the population structure has not been investigated. In this study, genetic diversity and population structure of the bronze featherback were examined using nucleotide sequence analysis of the mitochondrial DNA control region for 332 fish collected from Tonle Sap Lake and the Mekong River in Cambodia. The average nucleotide diversity (π) of all samples was 0.033, and the Mekong River samples had higher nucleotide diversity (0.034) than Tonle Sap Lake samples (0.028). The nucleotide diversity between the lake and river samples varied from 0.029 to 0.037. The genetic differentiation between the river and lake populations was also supported by the pairwise F -statistic values and hierarchical analysis of molecular variance, indicating that the Tonle Sap Lake population is genetically isolated from the population in main stream of the Mekong River.     

源于鳞被相关基因的微卫星标记与鲤 4 种生长性状的相关性分析

选用来源于鲤基因组中的鳞被相关基因(ant、eda、edar、fgfr)及其上下游序列中的155个微卫星标记,对德国镜鲤(Cyprinus carpio L.)与黑龙江野鲤(Cyprinus carpio haematopterus)杂交的F2 116尾个体的遗传多样性进行检测,以卡方检验估计群体Hardy-Weinberg平衡.结果表明,36个微卫星标记表现为多态,各标记的等位基因数在2～4个浮动,共检测到86个等位基因,平均每个标记2.388 9个;平均有效等位基因数为2.209 4;平均观测杂合度为0.624 5;平均期望杂合度为0.529 2;平均多态信息含量为0.432 1,其中23个微卫星标记表现为中度多态(0.25≤PIC＜0.5),13个微卫星标记表现为高度多态(PIC≥0.5),说明这个群体属于中度多态性水平.Hardy-Weinberg平衡检验结果显示,61％标记显著偏离平衡,人工选择压力和自交对家系造成了严重的影响.SPSS 17.0分析发现分别有10(28％)、7(19％)、7(19％)和11个(31％)标记与体质量、体长、体高和体厚存在显著相关性,并发现11个优势基因型.源于鳞被相关基因的微卫星标记与生长性状连锁的比例较高,以上结果从分子水平上提示鳞被基因与所研究4种生长性状存在一定的相关性.     

A set of highly polymorphic microsatellites useful for kinship and population analysis in turbot (Scophthalmus maximus L.)

The turbot (Scophthalmus maximus) is a flatfish species of great commercial value for aquaculture. In this study we describe the isolation and characterization of 30 novel highly polymorphic microsatellite markers in this species obtained from a genomic library enriched for seven short tandem repeated motifs. Much higher polymorphism (mean number of alleles: 13.37; mean expected heterozygosity: 0.869) and potential for parentage assignment than previously reported for microsatellites in turbot were found after the analysis of 24 wild individuals. Most loci conformed to Hardy–Weinberg expectations, excluding Sma‐USC20 and Sma‐USC28, which showed a high heterozygote deficit probably due to the presence of null alleles. No significant genotypic disequilibrium was observed between any pair of loci, suggesting no close linkage between them. These loci are potentially useful for kinship and population analysis in turbot.     

Evaluation of genetic diversity of eight grouper species <Emphasis Type="Italic">Epinephelus</Emphasis> spp. based on microsatellite variations

ABSTRACT:   Genetic diversity among eight species of grouper, Epinephelus bleekeri , E. coioides , E. malabaricus , E. ongus , E. akaara , E. maculatus , E. merra and E. fuscoguttatus , was studied using six microsatellite loci, Em-01 *, Em-03 *, Em-07 *, Em-08 *, Em-10 *, and CA-07 *, with the aim of exploring the feasibility of using microsatellite data for species identification. The results showed high levels of genetic differentiation among species ( F _ST  = 0.4403 and R _ST  = 0.4954). Species identification based on fixed allelic differences was possible between E. coioides , E. ongus , and E. fuscoguttatus at Em-01 * and between E. fuscoguttatus and E. ongus at Em-08 *. Private alleles were found in all species, except for E. ongus . Pairwise F _ST ranged 0.238–0.578 ( P  < 0.008 Bonferroni correction), and Nei's genetic distance ranged 0.433–2.710. Size homoplasy was observed at Em-03 * 157 allele, which was characterized by a T–C transition at the 119th nucleotide site of PCR products. The genetic assignment test unambiguously assigned each individual to the correct species. Thus, this test can be used for species identification of unknown individuals when the multilocus genotypes of the six microsatellite loci are available. The phylogenetic (neighbor-joining) tree, which was constructed based on the genetic distance matrix, separated the eight grouper species into two main groups.     

泥蚶34个EST-SSR标记的开发及在格粗饰蚶中的通用性检测

利用泥蚶转录组高通量测序、拼接获得的大量EST序列开发SSR标记,在1123条EST序列里筛查到73条含有SSR位点的EST序列,其中54个位点适合设计引物,在位点两侧设计引物并进行PCR扩增.结果显示,46对引物获得稳定扩增的位点,引物在泥蚶奉化群体的多态性检测中发现,有34对引物表现出多态性,共扩增出122个等位基因,各位点的等位基因数Na为2～7个,平均每个位点产生3.59个等位基因,观测杂合度Ho、期望杂合度He、多态信息含量PIC范围分别为0.000 ～ 0.600、0.078 ～0.771、0.106～0.718;Hardy-Weinberg平衡检测显示,13个位点偏离了平衡状态;用Nr和Swiss-Prot蛋白质数据库对含有多态性SSR的EST进行了基因注释,25个SSR位点来自注释基因序列.将34对泥蚶多态性SSR引物在格粗饰蚶中进行了通用性检测,结果有1 1对成功扩增,8对表现为多态,通用率为23.53％,这些通用引物可用于两种蚶的遗传多样性评价、系统进化分析、比较作图和基因发掘等研究.