Parentage Determination and Effective Population Size Estimation in Mass Spawning Pacific Oyster,Crassostrea gigas,Based on Microsatellite Analysis

Seven high polymorphic microsatellite loci were used to determine the pedigrees in a mass spawning of Pacific oyster, Crassostrea gigas, and to estimate the genetic variability between broodstock and offspring. Parental assignment was performed on a total of 155 individuals, including 141 offspring, 8 candidate mothers, and 6 candidate fathers. The assignment results of real offspring were generally in agreement with simulation with a success rate over 99% using only six of these loci. The allelic diversity and observed heterozygosity (H_o) exhibited similarity between parents and offspring populations, but the expected heterozygosity (H_e) had a significant decrease in offspring. Although all the males and females contributed to the next generation, the variances of reproductive success and unequal sex ratio resulted in a decline in effective population size (N_e = 11.42). The inbreeding rate of this small‐scale, mass spawning population was estimated at approximately 16.5% per generation. This gave us an insight that when designing breeding programs based on mass spawning for future oyster cultivation generations, the higher inbreeding and lower effective population size should be considered.     

Genetic diversity status of Pacific white shrimp (<Emphasis Type="Italic">Litopenaeus vannamei</Emphasis>) using SSR markers in Iran

Genetic diversity is vital for the maintenance of genetic pool in cultured shrimps. In order to estimate the current status of genetic diversity in Litopenaeus vannamei shrimp in Iran, as an exotic species, a total of 45 individuals from Amiri and Gorgeaj farms in Jask port of Hormozgan province and one hatchery in Gomishan city of Golestan province, were detected using four microsatellite loci. The number of alleles per locus was 5–10, and the mean effective number of alleles (N _e) across populations and loci ranged from 4.834 to 5.148. The overall mean observed heterozygosity (H _o) ranged from 0.450 to 0.479, which was lower than the expected one (0.789–0.794). There was nothing remarkable about any of the allele frequencies across populations or loci. The mean inbreeding coefficient (F _IS) and pairwise genetic differentiation (F _ST) among populations were 41.6 % and 0.133, respectively. The three studied populations departed from Hardy–Weinberg equilibrium (HWE). Analysis of molecular variance revealed 14 % variability among and 86 % within populations. However, considering departing from HWE and the high F _IS and F, the moderate pairwise F _ST values, importance of introducing genetically diverse broodstock and monitoring to control inbreeding is discussed.     

High allozyme variation and genetic similarity of two populations of commercial penaeids, Penaeus brevirostris (Kingsley) and P. vannamei (Boone), from the Gulf of California

Genetic resources assessment is considered as the starting point for population management in both fisheries and aquaculture. In this study, we have obtained genetic variability measures of two commercial penaeid shrimp species, Penaeus brevirostris (Kingsley) and P. vannamei (Boone), from the Gulf of California by means of starch electrophoresis analysis of 16 enzymatic systems, plus total protein. Thirty-two loci were resolved, which revealed polymorphism values (P₉₅) of 15.63% in P. brevirostris and 25.00% in P. vannamei. Unbiased expected heterozygosity (H_e) was 0.076 and 0.105 respectively. These are high genetic variability figures, taking into account the previously low genetic variability, assessed by means of allozyme variation, reported in several penaeid species around the world. Nei’s genetic similarity between the two species is 0.891.     

Effects of artificial selection practices on loss of genetic diversity in the Pacific abalone,Haliotis discus hannai

The Pacific abalone, Haliotis discus hannai Ino, is one of most important aquaculture species in China. Artificial selection has been the common and inevitable approach in breeding. In present study, the effect of artificial selection on the genetic structure of two abalone lines (JJ selection strain, three successive selection lines based on the fast‐growing trait, which was named JJF0, JJF1, JJF2, and JJF3; and R selection strain, the offspring of the red shell colored variants) were evaluated using 10 microsatellites. Loci showed from low to high polymorphism, with the number of alleles (A) ranging from 2 to 18 in each population. The mean observed (H_o) and expected heterozygosities (H_e) were 0.650 ± 0.022 and 0.711 ± 0.018, respectively. In selection strain JJ, the values for most diversity genetic indexes (A_e, the number of effective alleles, H_o and H_e) decreased from JJF0 to JJF3. Meanwhile, compared to JJ line, the genetic diversity estimates of R were close to those of JJF3. Nei's genetic distance ranged from 0.20357 to 0.51346. The unweighted pair group method with arithmetic mean tree based on Nei's genetic distance also showed that the control Japan population and JJF0 formed to a cluster firstly, which were subsequently grouped together with JJF1, JJF2, JJF3, whereas the R was isolated from the rest of the populations. Analysis of genetic information indicated that genetic diversity was lost with artificial selection practices. Mechanisms underlying the maintenance of an acceptable level of genetic diversity while pursuing economic interests should be conducted in future research studies.     

Can genetic diversity be maintained during mass selection of the Chinese mitten crab,Eriocheir sinensis?

The Chinese mitten crab (Eriocheir sinensis) is an important aquaculture species and food source in Eastern Asia. This study assesses the changes in genetic diversity in successive generations of early‐ and late‐maturing strains of E. sinensis using 30 microsatellite markers with high polymorphism. The mean average number of alleles (N) in the founder population (G0), first generation (G1), second generation (G2) and third generation (G3) of the early‐maturing strain were 18.367, 14.800, 16.400 and 16.533, respectively; while in late‐maturing strain the values were 18.500, 16.267, 14.367 and 16.533 respectively. Likewise, there was a slight decline in average allelic richness (R_s) in the three successive generations. In both strains, the mean observed heterozygosity (H_o) remained relatively constant for the early‐maturing strain and the values were 0.655, 0.667, 0.685 and 0.705, respectively; and for the late‐maturing strain these were 0.665, 0.672, 0.688 and 0.702 respectively. Similarly, the expected heterozygosity (H_e) remained constant, ranging from 0.823 to 0.854. There was a decrease in effective population sizes (N_e) of the early‐maturing strain with successive generations: values were 492.2, 35.0, 134.7 and 193.2, respectively; while the values in the late‐maturing strain were 1268.5, 75.6, 111.5 and 97.2 respectively. All pairwise population distances were very close in both strains. In conclusion, these results suggest that mass selection of E. sinensis did not significantly diminish genetic diversity although there was a decline for the N_e. Therefore, it is important to maintain sufficient broodstock numbers and a large effective population when following a selective breeding programme.     

基于SSR-seq技术评估中间球海胆多代选育群体和普通养殖群体的遗传多样性

为明确不同选育群体中间球海胆的遗传多样性和遗传结构,利用SSR-seq技术和15个微卫星位点,对1个家系选育群体(FP)、1个群体选育群体(IP)和1个未经选育的普通养殖群体(CP)的遗传多样性及遗传结构进行了分析。结果显示,15个微卫星位点共检测出112个等位基因,FP、IP、CP 3个群体的平均观测等位基因数(N_a)分别为5.077、5.133和6.133个,平均有效等位基因(N_e)分别为2.816、2.873和3.638个,平均观测杂合度(H_o)分别为0.522、0.441和0.501,平均期望杂合度(H_e)分别为0.595、0.599和0.667,平均多态性信息含量(PIC)分别为0.546、0.543和0.623。家系选育群体(FP) H_e与H_o的差值(0.073)低于IP (0.158)和CP (0.166),平均固定指数(F)(0.115)低于IP (0.248)和CP (0.246)。3个群体间遗传分化系数(F_st)介...     

Genetic structure and genetic relatedness of a hatchery stock of Senegal sole (Solea senegalensis) inferred by microsatellites

Solea senegalensis is a fish species in an early stage of domestication. A commercial hatchery in southern Spain experienced a dramatic failure in spawning and embryo survival, after incorporation into the broodstock of fish reared in their hatchery. To assess the impact of management on the stock quality, a population genetics study was made of adult broodstock present at this hatchery in 2000. The broodstock was composed of a group of fish of wild origin and two more groups from F₁ progeny. A set of eight microsatellite loci was used to compare genetic variability among the three groups and to establish the relationships between pairs of individuals within each group.Individuals from F₁ origin showed a substantial reduction in genetic variability when compared to those from the wild. There was a greater than 50% decrease in the number of alleles per locus corrected by sample size and 16% and 26% reductions in H_e values in the two F₁ groups, respectively. The degree of genetic relationship between individuals, evaluated by the relatedness estimator, rxy, and by graphical approaches, revealed a high proportion, almost 75%, of full-sibs and half-sibs in both F₁ groups, which might account for the reduction in overall variability. The high proportion of siblings also suggests a reproductive pattern in this stock in which few individuals breed successfully. This mating behaviour, along with factors inherent to selection and management systems, might be responsible for the negative effects on the reproductive capacity of the stock.     

福瑞鲤与豫选黄河鲤选育群体的遗传结构及亲本间遗传距离分布

用微卫星标记分析了鲤鱼(Cyprinus carpio L.)的2个品种福瑞鲤和豫选黄河鲤选育群体的遗传结构,并揭示了雌雄个体间遗传距离的分布规律。结果表明,23个微卫星标记在福瑞鲤(FR,n=192)和豫选黄河鲤(YX,n=96)中各检测到160个和131个等位基因。福瑞鲤的平均有效等位基因数(N_e)、观测杂合度(H_o)、期望杂合度(H_e)和多态信息含量(PIC)分别为4.559、0.695、0.741和0.702,群体处于高度多态水平(PIC≥0.5);豫选黄河鲤的4项遗传多样性参数分别为3.620、0.665、0.642和0.600。虽然豫选黄河鲤同样处于高度多态水平(PIC≥0.5),但是N_e、H_e和PIC均极显著低于福瑞鲤(P0.01),说明福瑞鲤的杂交选育背景决定了其较系统选育的豫选黄河鲤具有较多的来源于不同亲本的等位基因;而两者H_o差异不显著(P0.05),说明豫选黄河鲤种内也保持了较高的遗传杂合度。分别统计福瑞鲤与豫选黄河鲤雌雄个体间的遗传距离,结果表明两两雌雄个体间遗传距离呈正态分布。福瑞鲤个体间遗传距离的中间值位于0.8~1.0,占37.39%;而豫选黄河鲤个体间遗传距离中间值位于0.5~0.7,占49.33%。建议福瑞鲤和豫选黄河鲤在家系配组时,选择亲本间遗传距离阈值范围在0.8~1.0和0.5~0.7为宜。     

微卫星用于凡纳滨对虾育种过程中亲权分析及遗传多样性的变化监测

利用7个微卫星标记分析了6个凡纳滨对虾家系的亲本（G₀）及其后代（G₁）的基因型分离情况,同时对G₀和G₁的所有座位期望杂合度进行配对比较分析,并且通过对G₁群体每个位点的F-statistics分析检验群体的遗传分化,以期对凡纳滨对虾育种进行遗传监测。结果表明：共检测到44个等位基因。G₀和G₁的平均每个座位的等位基因数目分别为6.14和6.27,平均期望杂合度为0.786和0.733,平均多态性信息含量为0.709和0.695。7个微卫星座位的累计排除概率为0.99,并且在有亲本存在的情况下,能够将6个家系分开。G₀与G₁的所有座位期望杂合度的配对比较分析结果表明G₀平均期望杂合度要显著高于G₁(P<0.01)。G₁各座位的遗传分化指数F_ST在0.270 3～0.465 4,平均分化系数为0.358 4。说明亚群间属于高度遗传分化。最后,根据6个家系的遗传距离,利用UPGMA法对G₁个体聚类分析,结果表明亲缘关系较近的家系A和B以及C和D的相似系数很高分别各聚为一类,E和F分别聚为一类。     

利用微卫星荧光多重PCR技术分析壳白长牡蛎3代人工选育群体的遗传多样性

为了探讨壳白长牡蛎人工选育对群体遗传变异的影响,实验利用4个多重PCR组合共10个微卫星标记分析了连续3代壳白长牡蛎人工选育群体和野生群体及基础群体的遗传多样性。结果发现,6个群体的平均等位基因数量为7.2~12.6,等位基因丰度为6.8~11.0,期望和观测杂合度分别为0.672~0.769和0.486~0.542;与野生群体相比,3代选育群体的平均等位基因数显著降低,但平均期望杂合度并无显著差异。哈迪—温伯格平衡检验结果显示,在60个群体—位点组合中有39个群体—位点组合显著偏离哈迪—温伯格平衡,近交系数F_(is)范围为0.215~0.342。群体间遗传分化指数F_(st)范围为0.005~0.076,处于中—低等的遗传分化水平。研究表明,虽然连续选育对群体的遗传多样性和遗传分化造成了一定程度的影响,但人工选育群体依然表现为较高的遗传多样性,仍可以一定的选择压力对选育群体进行人工选育。     

棘头梅童鱼七个野生群体遗传多样性的微卫星分析

为研究中国沿海地区棘头梅童鱼群体的遗传多样性,利用微卫星标记技术,采用9对微卫星引物对中国连云港(LYG)、大丰(DF)、崇明(CM)、舟山(ZS)、温州(WZ)、宁德(ND)、厦门(XM)棘头梅童鱼7个野生群体的遗传多样性和遗传结构进行了分析。结果显示,实验检测到63个等位基因,各位点等位基因数为3~13个,有效等位基因数为1.7510~8.0317;各位点的观测杂合度(Ho)为0.3596~0.7854,期望杂合度(He)为0.4300~0.8780;多态信息含量(PIC)为0.3604~0.8631,其中有2个位点表现为中度多态(0.25PIC0.5),7个位点表现为高度多态(PIC0.5),具有较丰富的遗传多样性水平。Hardy-Weinber平衡分析显示,7个群体的大部分位点未偏离平衡。基于群体间Nei氏标准遗传距离构建的7个野生群体UPGMA系统进化树结果显示,ND和WZ群体遗传关系最近,ZS和WZ群体遗传关系最远,WZ和ND聚为一支,但总体上没有显著的遗传分化。     

Conservation genetics of the Mary River turtle (Elusor macrurus) in natural and captive populations

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Genetic characterization of naturalized populations of brown trout Salmo trutta L. in southern Chile using allozyme and microsatellite markers

This study describes the genetic structure of five naturalized populations of brown trout in southern Chile using allozyme and microsatellite markers to establish levels of intra‐ and interpopulation genetic variability and divergence. Fourteen enzymatic systems were used comprising 20 loci and three microsatellite loci specific to brown trout. The genetic variability values (allozymes, P=20–35%, average=27%, H_O=0.118–0.160, average=0.141; microsatellites, P=33.3–100%, average=66.66%, H_O=0.202–0.274, average=0.229) are similar to values described in other naturalized populations of brown trout present in Chile, but higher than those observed in European populations of this species. Values of total genetic diversity (H_T) (allozymes=0.1216 and microsatellites=0.3504) and relative genetic divergence (G_ST) (allozymes=9.5% and microsatellites=15%) were also similar to the results obtained in previous studies of Chilean populations of brown trout. These values, when compared with those obtained in Europe, proved to be similar for H_T but lower for G_ST. The low interpopulational genetic differentiation was in accordance with the small genetic distance observed between the populations analysed (D Nei=0.004–0.025). On the other hand, the high frequency of one of the two alternative alleles of the phylogeographic marker locus LDH‐5* in the populations analysed (LDH‐5*90>0.84) would indicate a European origin, in particular Atlantic as opposed to Mediterranean, for the brown trout introduced into Chile. The high levels of genetic variability suggest a mixed origin for the naturalized brown trout in Chile, which could have originated either before or during the introduction process. Nevertheless, the low level of genetic differentiation between populations could reflect the short lapse of time in evolutionary terms, during which populations introduced into Chile have been exposed to different evolutionary forces, and which has not been sufficiently long to produce greater genetic differentiation between populations.     

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.     

Polymorphic microsatellite differences among four cultured populations of two selected tilapia strains

Genetic diversity plays a vital role for maintaining economically important traits in fish populations. The goal of this study was to assess the genetic variability among four tilapia populations in China. Two populations named XA (Oreochromis aureus, 30 samples) and AN (Egyptian Oreochromis niloticus, 24 samples) were gathered from Freshwater Fisheries Research Center (FFRC), and the other two named AL (O. aureus, 30 samples) and AJ (Egyptian O. niloticus, 30 samples) were from Guangxi Fisheries Research Institute (GFRI). GFRI introduced AL and AJ from XA and AN, and selected them for generations. We want to know if AL and AJ have genetic changes after selection and generations. Fifteen polymorphic microsatellite markers were used. Our results indicated that the four tilapia populations showed moderate genetic variability. The polymorphism information content (PIC) of the four populations was from 0.28 to 0.39; A_e was from 1.67 to 2.15; the H_e was from 0.35 to 0.46; the genetic similarity indexes were from 0.94 to 0.69 and also the clustering result showed that these four populations were highly related. There were no significant changes in genetic variation between late‐introduced population and early‐introduced population of O. aureus. Our results helped to understand the genetic variability of tilapia populations in China and for future marker‐assisted selection for important economic traits.     

Genetic diversity and relatedness estimates for captive barramundi (Lates calcarifer,Bloch) broodstock informs efforts to form a base population for selective breeding

Aquaculture of barramundi or Asian seabass (Lates calcarifer) is growing in both Australia and Southeast Asia and there is substantial interest to improve production efficiency through selective breeding. The establishment of a large and genetically diverse base population is a prerequisite for a sustainable and long‐term productive breeding program. Before selective breeding programs can begin for Australian barramundi it is important to assess the overall genetic diversity of current captive broodstock populations. To address this question, 407 captive barramundi broodstock from eight separate Australian broodstock populations were genotyped using 16 polymorphic microsatellite DNA markers. A Bayesian STRUCTURE analysis indicated that captive Australian broodstock are broadly divided into two genetic stocks. Multivariate analysis between broodstock individuals and pairwise F_ST between broodstock populations also supported the existence of two stocks. Comparisons with data obtained from natural stocks suggested that hatchery individuals were either sourced from the two stocks or represented an admixture between them. Genetic diversity was low within each broodstock population (allelic richness ranged from 2.67 to 3.42 and heterozygosity ranged from 0.453 to 0.537) and relatedness estimates within hatcheries were generally low (average r was equal to 0.141). We recommend sourcing captive individuals according to high levels of neutral genetic diversity and low levels of relatedness for the establishment of a base population. We also make recommendations about including genetically diverse wild individuals.     

Conservation genetic assessment of the paleback darter,Etheostoma pallididorsum,a narrowly distributed endemic in the Ouachita Highlands,Arkansas, USA

1. The paleback darter, Etheostoma pallididorsum, is considered imperilled and has recently been petitioned for listing under the Endangered Species Act. Previous allozyme-based studies found evidence of a small effective population size, warranting conservation concern. The objective of this study was to assess the population dynamics and the phylogeographical history of the paleback darter, using a multilocus microsatellite approach and mitochondrial DNA.
2. The predictions of this study were that: paleback darter populations will exhibit low genetic diversity and minimal gene flow; population structure will correspond to the river systems from which the samples are derived; reservoir dams impounding the reaches between the Caddo and Ouachita rivers would serve as effective barriers to gene flow; and the Caddo and Ouachita rivers are reciprocally monophyletic.
3. Microsatellite DNA loci revealed significant structure among sampled localities (global F_st = 0.17, P < 0.001), with evidence of two distinct populations representing the Caddo and Ouachita rivers. However, Bayesian phylogeographical analyses resulted in three distinct clades: Caddo River, Ouachita River, and Mazarn Creek. Divergence from the most recent ancestor shared among the river drainages was estimated at 60 Kya. Population genetic diversity was relatively low (H_e = 0.65; mean alleles per locus, A = 6.26), but was comparable with the population genetic diversity found in the close relatives slackwater darter, Etheostoma boschungi (H_e = 0.65; A = 6.74), and Tuscumbia darter, Etheostoma tuscumbia (H_e = 0.57; A = 5.53).
4. These results have conservation implications for paleback darter populations and can be informative for other headwater specialist species. Like other headwater species with population structuring and relatively low genetic diversity, the persistence of paleback darter populations is likely to be tied to the persistence and connectivity of local breeding and non-breeding habitat. These results do not raise conservation concern for a population decline; however, the restricted distribution and endemic status of the species still renders paleback darter populations vulnerable to extirpation or extinction.

     

Loss of genetic variability in the captive stocks of tambaqui,Colossoma macropomum (Cuvier, 1818), at breeding centres in Brazil,and their divergence from wild populations

The loss of variability in farmed populations and the risks of interactions with wild populations support the need for the genetic monitoring of species farmed throughout the world. In Brazil, the tambaqui is the most widely farmed native fish species. Despite this, there are no data on the pedigree of the farmed stocks, and the potential for interactions with wild populations in the Amazon basin has raised concerns with regard to the genetic variability of these stocks. The present study analysed sequences of the mitochondrial Control Region and 12 microsatellites to characterize the genetic variability of seven historically important commercial tambaqui breeding centres located in four different regions of Brazil, and compared these sequences with those obtained from individuals collected from a wild population. High levels of genetic diversity were found in the wild population, whereas genetic diversity was reduced in both markers in most captive populations, except for the broodstock located near the Amazon River. High F_ST and D_EST indices were recorded between the wild population and most of the captive stocks analysed. The drastic reduction in genetic diversity found in most captive stocks and the difference between these stocks and the wild population may have been the result of the small size of the founding populations and the absence of breeding management. The renewal of the broodstocks and the application of breeding management techniques are recommended. In the Amazon region, in addition, the use of broodstocks that are genetically very different from local wild populations should be avoided.     

利用高通量测序开发的熊本牡蛎微卫星标记评价野生和养殖群体的遗传多样性及通用性

利用高通量测序的方法,从熊本牡蛎基因组中开发了20对具有多态性的微卫星标记,通过微卫星标记位点比较了野生群体和养殖群体的遗传多样性。野生群体中,所有位点共扩增出330个等位基因,等位基因数(N_a)范围为6～39,平均等位基因数为16.500 0;有效等位基因数(N_e)范围为1.352 9～33.361 7,平均值9.517 2;观测杂合度(H_o)范围为0.200 0～1.000 0,平均值0.671 5;期望杂合度(H_e)范围为0.265 6～0.987 7,平均值0.832 1;ShannonWeiner指数(Ⅰ)范围为0.648 3～3.585 8,平均值2.276 9;多态信息含量(PIC)范围为0.254 5～0.969 2,平均值0.803 5,共有16个位点符合Hardy-Weinberg平衡。养殖群体中,N_a平均值为10.250 0,N_e平均值为5.843 4,H_o平均值为0.639 1,H_e平均值为0.763 6,I平均值为1.791 4,PIC平均值为0.720 7。结果显示,熊本牡蛎养殖群体的遗传多样性低于野生群体,但仍然维持在高度多态水平。研究表明,在熊本牡蛎人工繁育过程中,使用大数量的亲本进行繁育,可有效防止选育群体的遗传多样性降低,但人工选育对选育群体的遗传多样性也产生了一定的影响。另外,分析了这些引物在近缘种葡萄牙牡蛎、长牡蛎、香港牡蛎、有明牡蛎、僧帽牡蛎、咬齿牡蛎以及舌骨牡蛎中的通用性情况,发现XB1-6、XB1-39和XB1-45 3个位点在8个物种中均能扩增出目的条带,XB1-41仅能在熊本牡蛎中扩增出目的条带。     

Development of 25 Novel Microsatellite Loci and Genetic Variation Analysis in Breeding Populations of the Pearl Oyster,Pinctada fucata

In this study, we screened 103,050 gene sequences of Pinctada fucata and identified 1450 simple sequence repeats (SSRs) distributing in 1355 sequences, in which 97 have sufficient flanking sequences for primer design, and 25 novel microsatellite loci were stable and polymorphic with N_a which varied from 2 to 6. Six microsatellite loci were found in the open reading frame of their corresponding sequences. With the 25 SSR loci, five breeding populations and a wild population, a total of 240 individuals from wild population (W), three consecutive selected populations (G₁, G_2, and G₃), and two backcross populations (D_o, D_r) (40 individuals per population) were assayed. In these populations, we detected 108 alleles of 111 bp to 362 bp. Each population had mean H_o of 0.5718–0.7366 and mean H_e of 0.5830–0.6954. Except the F_st = 0.0697 (W vs. G₃), pairwise F_st values ranged from 0.0131 to 0.0473, the genetic identity coefficients varied from 0.8688 to 0.9663, and genetic distance ranged from 0.0343 to 0.1307, all of these suggested that the breeding populations had genetic differentiation but at a low level generally. Besides, all the genetic index values, backcross populations (D_o, D_r) were higher than corresponding G₃ values, indicating that backcross could make the offspring incline to the recurrent parents.