Evaluation of genetic characteristics of wild and cultured populations of the Japanese pearl oyster <Emphasis Type="Italic">Pinctada fucata martensii</Emphasis> by using AFLP markers

Genetic characteristics of four wild (Mie, Fukui, Shimane and Nagasaki) and five cultured populations (selectively bred for 12 years with an origin of Ehime population) of the Japanese pearl oyster, Pinctada fucata martensii, were evaluated by using AFLP markers. Six primer pairs generated 1019 loci in total, among which 45.2–55.1 % was polymorphic among populations. Although there was no significant difference in gene diversity between wild (0.170–0.174) and cultured (0.158–0.173) populations, genetic relatedness in cultured populations (0.316–0.450) was about three times higher than that in wild populations (0.110–0.165). In addition, genetic differentiation was about twenty times larger in cultured populations (Nei’s distance: 0.0111) than in wild populations (Nei’s distance: 0.0005). These results mean that selective breeding can cause marked inbreeding as well as large genetic differentiation among cultured populations in a short period. On the other hand, it was suggested that genetic homogenization in the wild, probably due to a large-scale transport of cultured oysters, had progressed in the sea around Japan. In conclusion, it is necessary to prevent inbreeding by the reconsideration of the style of selective breeding in cultured populations, while the escape of gametes or spats of cultured strains in the wild should be avoided for the preservation of genetic characteristics in native populations of P. f. martensii.     

Effect of water temperature and chlorophyll abundance on shell growth of the Japanese pearl oyster, Pinctada fucata martensii, in suspended culture at different depths and sites

To understand the relationships between shell growth and some environmental factors, we examined the relationships between water temperature or chlorophyll abundance and the shell growth of the Japanese pearl oyster, Pinctada fucata martensii, suspended at three different depths at two sites. Growth in height, length and thickness of the shells were limited by water temperature during winter (< 20 °C), whereas growth in thickness correlated with food abundance, measured as chlorophyll, during early summer (> 20 °C). These results suggest that the shell of P. fucata martensii could grow well at locations with greater abundance of food and adequate water temperatures (20–26 °C), resulting in a longer growing season.     

Genetic assessment of milkfish (Chanos chanos Forsskal) stocks based on novel short tandem repeats for marker‐aided broodstock management

Milkfish hatchery broodstock are either from on‐grown wild‐caught or hatchery‐produced fry/juveniles. To determine if a marker‐assisted management scheme can be formulated for improved milkfish hatchery production, milkfish stocks were genetically characterized using nine novel short tandem repeats or microsatellites. Eight wild‐bred Philippine stocks (CLA, CUR, CAM, SIH, SBH‐I1, HH, PAL and ZH‐P₀), four hatchery‐bred stocks (SBH‐I2, SBH‐D, BoH and ZH‐F₁), two farm stocks of known mixed lineages (SPH and BDH) and one Indonesian hatchery‐bred stock (WJH) were assessed. WJH was included since milkfish fingerlings from Indonesia reared in Philippine farms could be developed into future broodstock. Mean allelic richness (Ar) was highest in wild‐bred stocks (9.5) and lowest in hatchery‐bred spawners (9.1). Mean expected heterozygosities (He) were relatively similar in all stocks with wild‐bred stocks slightly higher (0.67) than the others. An analysis of molecular variance indicated significant yet low genetic differentiation among stocks (F_ST = 0.013; p = .000) where variation (98.6%) was explained by intra‐stock differences. In some of the domesticated stocks, reductions in mean allelic richness were observed in first generation hatchery broodstock (e.g. ZH‐F₁; Ar = 8.3), compared with their founder stock (e.g. ZH‐P₀; Ar = 9.4). The Indonesian stock was similar to local wild‐bred stocks based on genetic variability indices; thus, it might be likely that the local stocks’ fitness traits could be comparable with the imported milkfish stock which has been perceived to be better. The quality of locally available farmed milkfish and prospects of formulating a broodstock management scheme for the production of good quality milkfish seedstock are herewith discussed.     

Identification of differential broodstock contribution affecting genetic variability in hatchery stocks of Atlantic salmon (Salmo salar)

Supportive breeding of Atlantic salmon (Salmo salar) is commonly employed to maintain numbers of fish where the species has become locally endangered. Increasingly, one of the main aims of population management is the preservation of natural genetic diversity. If the stocks employed in supportive breeding exhibit reduced variation they can alter the natural pattern of genetic variation observed in wild populations. In northern Spain, wild adult salmon are caught every year from local rivers and artificially crossed in order to create supportive stocks. The offspring are hatchery reared until the juvenile stage, then released into the same river where their parents were caught. In the current study, our findings demonstrate that although adult broodstock exhibit a pattern of variation similar to the wild populations, variability at microsatellite loci was drastically reduced in the juveniles released into one of three rivers analyzed. The contribution of broodstock to this juvenile stock was examined by pedigree analysis. A restricted number of females contributing to the hatchery stock was identified as the main cause of loss in genetic variation, possibly due to overmaturity of some multi-sea-winter females. We suggest that better monitoring and control of parental contribution will help in solving the problem of loss of genetic diversity in hatchery populations.     

Impacts of hatchery release on genetic structure of rock carp <Emphasis Type="Italic">Procypris rabaudi</Emphasis> in the upper Yangtze River,China

Rock carp Procypris rabaudi is a vulnerable endemic fish in the upper Yangtze River. Hatchery release has been carried out as a major stock enhancement strategy for this species. Ten microsatellite loci were chosen to compare genetic variation between one wild population and two hatchery groups to evaluate the potential impacts of hatchery release on the genetic structure of the wild population. Two different models indicated strong evidence of recent bottlenecks in all groups. The hatchery groups were lower in the mean number of alleles per locus, allelic richness, and allelic diversity compared with the wild population. The 80% membership coefficient indicated that 14% of the wild fish could be assigned as hybrids of wild and hatchery fish. Our results suggested that hatchery release will further reduce the natural genetic diversity in the wild population, change the genetic structure of the rock carp population, and may not benefit restoration of this vulnerable fish species.     

基于线粒体Cytb基因和D-loop区序列的高邮湖湖鲚(Coilia nasus)遗传多样性分析

本研究利用线粒体细胞色素b(Cytb)基因和控制区(D-loop)序列作为分子标记,调查了高邮湖湖鲚(Coilia nasus)种群遗传多样性。结果显示,Cytb基因和D-loop区序列碱基A+T含量均高于G+C含量,显示碱基组成具有偏倚性。38条Cytb基因序列检出26个变异位点,定义13个单倍型,单倍型多样性和核苷酸多样性分别为0.716±0.078和0.002 70±0.000 57;40条D-loop区序列检出53个变异位点,定义21个单倍型,单倍型多样性和核苷酸多样性分别为0.906±0.034和0.006 27±0.000 99。13个Cytb基因单倍型之间的遗传距离在0.001~0.014之间,NJ系统进化树显示单倍型聚为1支;21个D-loop区单倍型之间的遗传距离在0.001~0.019之间,NJ系统进化树显示单倍型聚为2支。中性检测结果和歧点分布图均表明高邮湖湖鲚种群稳定,近期没有发生种群扩张。整体来看,高邮湖湖鲚种质资源遗传多样性水平较高,具有高单倍型多样性和低核苷酸多样性的特征。     

Genetic variation in farmed populations of the gilthead sea bream Sparus aurata in Greece using microsatellite DNA markers

Genetic variation in seven reared stocks of gilthead sea bream Sparus aurata, originating from Greek commercial farms, was assessed using five polymorphic microsatellite markers and was compared with that of two natural populations from the Ionian and the Adriatic Seas. The total number of alleles per marker ranged from 11 to 19 alleles, and hatchery samples showed the same levels of observed heterozygosity with samples from the wild but substantially smaller allelic diversity and expected heterozygosity. The global genetic differentiation for the cultivated samples was significant as indicated by F_st analysis, which might indicate random genetic drift and inbreeding events operating in the hatcheries. On the contrary, no significant difference was found between the two wild populations. Population pairwise tests between farmed and wild stocks were also significant, with the exception of one hatchery sample, the Central Greece 1, which was not significantly different from the two wild samples perhaps due to its recent use in aquaculture from wild‐caught animals. The UPGMA tree topology grouped the wild samples together with the Central Greece 1 stock, and showed a clear division between wild and farmed sample sets for the six remaining hatchery samples. Knowledge of the genetic variation in S. aurata cultured populations compared with that in the wild ones is essential for setting up appropriate guidelines for the proper monitoring and management of the stocks either under traditional practices or for the implementation of selective breeding programmes.     

Genetic diversity and population history of Tanichthys albonubes (Teleostei: Cyprinidae): Implications for conservation

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高邮湖大银鱼、太湖新银鱼Cytb和COⅠ 基因序列多态性分析

为探明高邮湖大银鱼、太湖新银鱼野生资源状况,利用线粒体DNA Cytb和COⅠ基因序列,对高邮湖大银鱼、太湖新银鱼的遗传多样性水平及遗传结构进行分析。试验结果显示,大银鱼Cytb基因序列全长1141 bp,其中多态性位点14个,共定义12个单倍型,单倍型多样性和核苷酸多样性分别为0.871±0.031和0.00172±0.00019,具有高单倍型多样性和低核苷酸多样性特征。COⅠ基因片段长度为630 bp,其中多态位点5个,共定义6个单倍型,单倍型多样性和核苷酸多样性分别为0.747±0.041和0.00202±0.00019,具有高单倍型多样性和低核苷酸多样性特征;太湖新银鱼Cytb基因序列全长1141 bp,其中多态性位点13个,共定义9个单倍型,单倍型多样性和核苷酸多样性分别为0.609±0.078和0.00094±0.00027,具有高单倍型多样性和低核苷酸多样性特征。COⅠ基因片段长度为630 bp,其中多态位点2个,共定义3个单倍型,单倍型多样性和核苷酸多样性分别为0.232±0.085和0.00038±0.00014,呈现低单倍型多样性和低核苷酸多样性。大银鱼和太湖新银鱼Tajima′s D和Fu′Fs中性检验值为负值,且歧点分布曲线呈单峰型,表明历史上经历过种群扩张。研究结果表明,应通过多种措施加强高邮湖银鱼种质资源保护。     

Development of Gene‐derived SNP Markers and Their Application for the Assessment of Genetic Diversity in Wild and Cultured Populations in Sea Cucumber,Apostichopus japonicus

The Apostichopus japonicus is a valuable aquaculture species in China. In this study, 51 single nucleotide polymorphisms (SNPs) were identified from expressed sequence tags of sea cucumber using high‐resolution melting. The average observed heterozygosity (Ho) and expected heterozygosity (He) were 0.2462 and 0.2897, respectively. Thirty‐two of these loci were used for estimating the genetic similarity and variation between the five hatchery stocks from China and two wild stocks from Japan. No significant differences in Ho or He were observed between the wild and hatchery populations. The pairwise Fst (which ranged from 0.0119 to 0.0236) and the genetic identity (which varied from 0.9802 to 0.9915) showed no significant differentiation between the wild and cultured stocks. The analysis of molecular variance indicated the source of variation was at the level of “within the populations.” The information on the genetic variation and differentiation in cultured and wild populations of A. japonicus obtained in this study is useful for setting up suitable guidelines for founding and maintaining of cultured stocks and for future genetic improvement by selective breeding.     

Effects of stocking on the genetic structure of brown trout,Salmo trutta,in Central Europe inferred from mitochondrial and nuclear DNA markers

Abstract Stocking has had a considerable effect on wild brown trout, Salmo trutta L., populations throughout Europe. To elucidate this impact and to outline further management strategies, the genetic structure of 25 wild populations and five hatchery stocks from Czech Republic and Slovakia were analysed using mitochondrial (control region) and nuclear DNA (microsatellites, LDH‐C1*) markers. Stocking practices have caused massive hybridisation between the Atlantic and Danube brown trout strains in the central Danube basin and have lead to a loss of among‐population divergence in Slovakia and the eastern part of Czech Republic. Comparison with studies from neighbouring countries revealed substantial differences in haplotype, allele frequencies and genetic diversity across Central Europe. Differences in stocking management and origin of breeding stocks appear to be crucial factors for the spatial variability of the genetic structure of brown trout.     

基于CO I和Cyt b序列的丹江口水库鲢群体遗传结构特征分析

有效的人工增殖放流应监控放流群体和野生群体的遗传结构特征,以避免放流群体对天然群体遗传多样性的负面影响。本研究利用线粒体CO I和Cyt b基因分析了丹江口鲢库区、鲢亲本和鲢子代3 个群体的遗传结构特征。分析结果显示,104 条646 bp线粒体CO I序列中共检测到多态位点15 个,简约信息位点5 个,单一变异位点10 个,定义了7 个单倍型,单倍型多样性为0.544~0.676,核苷酸多样性为0.00221~0.00254;103 条1058 bp线粒体Cyt b序列中共检测到多态位点19 个,简约信息位点13 个,单一变异位点6 个,定义了14 个单倍型,单倍型多样性为0.609~0.714,核苷酸多样性为0.00262~0.00424,总体上处于较高单倍型多样性和较低核苷酸多样性。CO I和Cyt b序列遗传距离、遗传分化指数以及基因流分析结果显示,群体间遗传距离为0.002（CO I）、0.003~0.004（Cyt b）,总的遗传分化指数为-0.00468（P>0.05）（CO I）、0.03180（P>0.05）（Cyt b）,差异均不显著,群体间基因流为14.69～41.47（CO I）、5.49～40.47（Cyt b）,分子方差分析（AMOVA）结果表明群体的遗传变异主要来自群体内。单倍型聚类关系树表明,3 个鲢群体间均存在共享单倍型,不同地理群体间单倍型散乱分布于各支,未形成地理群体的聚集。以上分析结果显示,3 个鲢群体间不存在明显的遗传分化,表明增殖放流鲢群体与丹江口库区野生群体的遗传多样性和遗传结构相近,可开展增殖放流。本研究结果可为丹江口鱼类增殖站鲢群体的增殖放流提供科学依据。     

Respiration rates of the Japanese pearl oyster, Pinctada fucata martensii, feeding on Pavlova lutheri and Chaetoceros gracilis

We examined the relationship between algal food (Chaetoceros gracilis and Pavlova lutheri) density and the respiration rate of Japanese pearl oysters, Pinctada fucata martensii. The respiration rate increased consistently with food density, independent of algal species. However, the respiration rate decreased at food densities >10⁷ µm³ mL^?1, suggesting inactivation of the digestion system by the high‐food density. Our results suggested that eutrophicated or turbid environments may be unfavourable to the growth of P. fucata martensii.     

Forecasting the genetic influences of red drum stock enhancement in South Carolina

The South Carolina Department of Natural Resources (SCDNR) began stocking red drum, Sciaenops ocellatus (Linnaeus), in 1989 to augment the abundance of juveniles available for recreational harvest in South Carolina estuaries. While stock enhancement can help supplement wild populations under high fishing pressure, releasing hatchery‐raised fish into the wild also presents the risk of decreased genetic diversity. An individual‐based model (IBM) was developed to forecast the genetic influences of stocking on the wild spawning population to inform responsible stocking strategies. Model results indicated the SCDNR red drum stock enhancement programme should maintain mean contributions of stocked fish no greater than 30% per year class over a 45‐year stocking period, coupled with at least 10 effective breeders in the hatchery replaced annually, to maintain current levels of genetic diversity estimated in the wild population. The IBM is a useful tool for hatchery managers to guide responsible stock enhancement.     

Estimating the heritability for growth-related traits in the pearl oyster,Pinctada fucata martensii (Dunker)

Twelve paternal half-sib families (or 36 full-sib families) of the pearl oyster, Pinctada fucata martensii (Dunker), were produced according to the requirements of hierarchical genetic mating design. A total of 4320 individuals, aged 15 months, were measured for seven growth-related traits. Predicated upon the additive-dominance genetic analysis model, varying genetic variance components and then heritabilities of the growth-related traits of interest were estimated using analysis of variance. Results showed that seven growth-related traits had larger additive genetic variances (P<0.05); the dominance genetic variance of shell weight (SW) was smaller (P>0.05), the dominance genetic variances of other six traits were all larger (P<0.05). Narrow- and broad-sense heritabilities for the seven traits were, respectively, 0.64 ± 0.10 and 0.78 ± 0.12 for shell length, 0.49 ± 0.06 and 0.63 ± 0.09 for shell height, 0.38 ± 0.14 and 0.54 ± 0.16 for shell breadth, 0.41 ± 0.17 and 0.56 ± 0.11 for hinge length, 0.53 ± 0.11 and 0.68 ± 0.08 for body weight, 0.35 ± 0.07 and 0.55 ± 0.08 for tissue weight and 0.67 ± 0.10 and 0.75 ± 0.16 for SW. All heritability estimates were statistically significant (P<0.05). According to these results, the mass selection procedure is suggested for the breeding of P. martensii.     

Microsatellite genetic variation in wild and hatchery populations of the sea cucumber (Apostichopus japonicus Selenka) from northern China

Farming of the sea cucumber Apostichopus japonicus (Selenka) started 20 years ago and is still in rapid expansion in China. In order to assess the genetic status of both wild and cultivated stocks of this species, we used eight microsatellite markers to estimate the level of genetic diversity within five hatchery stocks and two wild populations of A. japonicus, and compared the degree of genetic differentiation between them. High levels of polymorphism were observed over all loci. The mean alleles and expected heterozygosities over the seven stocks were 10.4–12.3 and 0.735–0.783 respectively. The results of the microsatellite survey provide no evidence to show that hatchery practice of the sea cucumber in China to date has significantly affected the genetic variability of the cultured stocks. Significant differentiation was found between most pairs of the hatchery stocks and wild populations (F_st range: 0.008–0.036), and no obvious difference was detected between the wild populations (F_st=0.008). The information on the genetic variation and differentiation obtained in this study can be applied for future genetic monitoring of A. japonicus aquaculture stocks and will be useful for future genetic improvement by selective breeding, and for designing suitable management guidelines for these genetic materials.     

Monitoring of the genetic variability of the hatchery and recaptured fish in the stock enhancement program of the rare species barfin flounder <Emphasis Type="Italic">Verasper moseri</Emphasis>

ABSTRACT:   In relation to the stock enhancement program for barfin flounder, hatchery juveniles produced in 2001 were genotyped using microsatellite DNA markers (msDNA) and then released to natural waters. Subsequently, recaptured individuals, designated as 'tentative recaptured', were examined using msDNA. In order to evaluate the effectiveness of the stock enhancement program, pairwise F _ST and genic differentiation tests were used to estimate the genetic divergence between the wild samples, the hatchery broodstock, and the tentative recaptured samples. Analysis showed significant differentiation among these three groups. Pedigree determination by msDNA was used to establish the origin of the tentative recaptured individuals, in order to elucidate whether they were hatchery produced, 'real recaptured', or wild specimens. Wild individuals were not found. The effective population size of the real recaptured stock was very low ( N _e = 16.6). Equal family survivability was observed between the released and real recaptured stocks, indicating that the genetic variability of the released stock was maintained in the natural environment. Future broodstock management, breeding designs, and family contribution equalization of the offspring to be released will be required in this rare species to avoid unintended genetic differentiation between the wild population and the hatchery broodstock.     

凡纳滨对虾引进群体和养殖群体的PCR-RFLP分析

采用PCR-RFLP方法,对2008年引进的SPF凡纳滨对虾群体(G0)与2006年引进亲虾繁育的两代群体(G1、G2)的遗传结构进行了分析。使用AluI、TaqI、M boI、SpeI、SspI共5种限制性内切酶对线粒体DNA控制区进行单酶切分析,酶切得到的单倍型在2~4个之间,三个群体的复合单倍型类型分别有3个、7个和9个,没有为三个群体所共有的复合单倍型,复合单倍型的分布显示群体间差异显著。三个群体(G0、G1、G2)的核苷酸多样度π值逐渐增加,分别为0.033 3、0.128 6、0.134 9,复合单倍型多样度h分别为0.622 2、0.850 0、0.847 0,说明养殖群体的遗传多样性大于引进群体。聚类分析的结果表明随着养殖世代的增加,养殖群体与引进群体的差异呈增大的趋势。由此推测G0与G1的亲本存在较大的遗传差异,并且在使用引进群体作为亲本的育种过程中可能出现了种质混杂。     

基于线粒体Cyt b基因的虾夷扇贝群体遗传结构分析

本研究利用线粒体细胞色素b(Cyt b)基因标记对虾夷扇贝(Patinopecten yessoensis)6个群体(长岛底播增殖群、海洋岛底播增殖群、獐子岛底播增殖群、旅顺自然群、"獐子红"人工选育群以及日本青森陆奥湾群)进行种质状况评估,研究结果表明,6个群体120个个体的Cyt b基因序列共检测到20种单倍型,其中"獐子红"人工选育群单倍型最少,日本群体单倍型最为丰富,两者单倍型多样性指数分别为0.10000和0.88400。分子方差分析(AMOVA)发现,日本群体与中国群体间变异百分比为15.34%,明显高于作为一个基因池时群体间遗传变异水平;就中国组群而言,其83.41%的遗传变异来自于群体内部,组内群体间的变异只占1.52%,说明中国群体个体间的遗传变异远大于群体间的遗传变异。F_(st)分析显示,中国群体与日本群体之间发生了中等水平的遗传分化(0.07455~0.17895,Fst0.05)。以遗传距离矩阵构建群体间分子系统树(UPGMA树),拓扑结构图显示中国5个群体先聚为一支,再与日本群体聚为一支。由此可见,中国养殖区的虾夷扇贝与日本原产地群体间已出现明显的遗传分化,且虾夷扇贝中国群体的遗传多样性处于较低水平。本研究结果可为虾夷扇贝的种质资源保护与可持续开发利用提供理论依据。