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.     

Genetic diversity and differentiation of sea trout (<Emphasis Type="Italic">Salmo trutta</Emphasis>) populations in Lithuanian rivers assessed by microsatellite DNA variation

The genetic diversity and differentiation of sea trout were studied in three river basins in Lithuania: Akmena-Dane, Bartuva, and Nemunas. A total of 282 individuals were genotyped at eight microsatellite loci. A similar level of genetic diversity was found in all of the populations studied: mean allelic richness ranged from 3.64 to 5.03, and average expected heterozygosity ranged from 0.588 to 0.721. Significant genetic divergence was observed among the different river basins as well as between populations within the drainages. All pairwise F _ST values were highly significant, ranging from 0.027 to 0.197. The analysis of molecular variance showed rather weak hierarchical population structuring within the Nemunas basin, which may be explained by extensive gene flow among different river basins or, alternatively, reflect the influence of artificial breeding. Information on genetic diversity and differentiation of the Lithuanian sea trout populations will be useful for future management decisions.     

Genetic and morphological divergences between wild and captive‐bred populations of Salmo trutta abanticus

Salmo trutta abanticus is a non‐anadromous trout species native to Lake Abant and Seven Lakes in Turkey. A restocking programme by captive breeding was initiated in 1999 to support S. trutta abanticus population. Reared 2‐year‐old juveniles from randomly caught wild parental individuals in Maçka breeding farm were introduced into Lake Abant. We aimed to compare genetic and morphological divergences between wild‐ and captive‐bred populations using seven microsatellite loci and geometric morphometric measurements. A significant genetic and morphological divergences were detected between all population in F_st and canonical variate analysis based on geometric morphometric with 10 homolog landmark. Eighty‐six microsatellites alleles were recorded across loci. Number of private alleles, observed alleles and observed heterozygosity are statistically significant higher in Maçka captive‐bred population than Lake Abant and Seven Lakes populations. Of 42 tests, three departures from Hardy–Weinberg equilibrium were detected in all populations after Bonferroni correction. Two pairs of loci (Ssa85 – Str73 and Str73‐Str543) in Maçka, one pairs of loci (Ssa85‐Str73) in Abant and two pairs of loci (Ssa85‐Str60 and Str73‐Str543) in Seven Lakes populations show linkage disequilibrium. Population structure analysed with Structure software showed three genetic groups (?K = 3) in our studied populations. Relatedness estimates show higher mean relatedness values (r = 0.220 ± 0.230) for Maçka captive‐breed population than wild populations of Abant Lake and Seven Lakes (r = 0.140 ± 0.210 and r = 0.170 ± 0.200 respectively).     

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.     

Genetic Variations in Populations from Farms and Natural Habitats of Asian Green Mussel,Perna viridis,in Singapore Inferred from Nine Microsatellite Markers

The Asian green mussel, Perna viridis, has been widely cultured in Southeast Asia as delicious seafood depending on natural spat. We analyzed the genetic diversity and population structure of three populations (two from Singapore and one from Malaysia) from natural habitats and two from Singapore farms, by genotyping a total of 262 adult individuals using nine polymorphic microsatellites. In all five populations, high allelic (number of alleles A = 11.0–14.4,allelic richness Ar = 10.4–13.9) and gene diversity (expected heterozygosity He = 0.73–0.77) were observed, whereas deficiency of heterozygosity (inbreeding index f = 0.08–0.12) was detected in all populations. Genetic differentiation among populations was low (pairwise fixation index ranged from 0.003 to 0.088), but statistically significant. The usefulness of the information about genetic variations for genetic resource management of the populations in wild habitats and farms to keep genetic variation and setting up a breeding program of Asian green mussel was discussed.     

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 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.     

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。     

基于微卫星评估草鱼放流亲本对野生群体遗传多样性的影响

利用筛选的13对草鱼多态性微卫星标记,开展了2011至2015年长江中游草鱼亲本增殖放流对野生群体遗传多样性的影响评估。通过对各位点的遗传多样性分析,13个微卫星位点的多态信息含量为0.8622(0.657~0.950),基因多样度为0.8555(0.675~0.936)。15个群体的有效等位基因数为7.4503~10.1536,等位基因丰度为11.483~15.204,说明15个草鱼群体的遗传多样性水平总体较高。遗传分化指数分析表明,群体间不存在显著遗传分化(FST5%)。通过贝叶斯聚类分析和主成分分析可将草鱼群体分为4个组群,根据分组结果以及来源划分分别对草鱼群体进行AMOVA分析,发现遗传变异大部分来自于群体内个体间,组间及组内群体间的分化水平较低(FCT5%,FSC5%),与FST分析结果一致。研究表明,当前草鱼亲本增殖放流模式对野生群体遗传结构影响不明显。     

Inbreeding evaluation using microsatellite and its effect on growth traits in turbot,Scophthalmus maximus

Turbot is an economically relevant fish, and simultaneously at the risk of inbreeding depression in China. Microsatellite DNA markers were used to evaluate inbreeding level of a random turbot breeding population and further investigate its effects on growth traits. DNA based inbreeding coefficients (F_DNA) were ranged from 0 to 0.4124. Average inbreeding coefficient was 0.0621 with a standard deviation of 0.0658. Regression analysis results showed inbreeding had a very significant (p < .01) effect on harvest body weight (BW) and significant (p < .05) effect on harvest body long (BL). Inbreeding depression, expressed as the percent change in phenotype per 10% increase in F, was ?4.00% for BW and ?1.14% for BL. The inbreeding depression on high inbreeding coefficient (more than 10%) individuals was further analyzed. A set of significant (p < .05) and larger inbreeding depression was obtained, ?7.15% and ?2.49% for BW and BL respectively. This study demonstrates that inbreeding has a negative effect on growth traits and highlights the need of DNA markers to control inbreeding and maximize the genetic diversity in selection breeding programs.     

近交对长牡蛎幼虫和稚贝生长与存活的影响

分别以2010年通过2个野生个体进行交配获得的A01全同胞家系和2011年通过A01家系子代进行交配获得A02全同胞家系为亲本,在2013年6月采用同时建立全同胞交配子一代F1(F=0.250)、全同胞交配子二代F2(F=0.375)及设置对照组(F=0)的方法,研究在相同环境条件下,不同实验组的受精率与孵化率以及近交对长牡蛎幼虫期、稚贝期生长和存活的影响,并初步探讨近交代数与近交衰退的关系。结果发现,各组的受精率均在90%以上,除F2组外其余2组的孵化率也在90%以上;幼虫阶段,F1组和F2组的壳高与壳长均从12日龄出现衰退(近交衰退系数,inbreeding depression coefficient,IDC0),且F2组壳高的近交衰退系数均小于同日龄F1组壳高的近交衰退系数;F1组和F2组的存活率在整个幼虫期间均出现衰退,且F1组和F2组存活率的近交衰退系数均随着幼虫日龄的增加而逐渐减小。稚贝阶段,F1组和F2组的平均壳高在各日龄均表现出近交衰退(IDC0),且F2组壳高的近交衰退系数均小于相同日龄F1组壳高的近交衰退系数;3个实验组的平均壳长在整个稚贝阶段无显著性差异;F1组和F2组存活率的衰退在不同日龄始终存在(IDC0),且随着稚贝日龄的增加其衰退程度逐渐加大。研究结果为长牡蛎选择育种和遗传改良提供了基础资料。     

草鱼野生与选育群体遗传变异微卫星分析

为探究经过2个选育世代后选育群体遗传多样性和遗传结构的变化,实验采用多重PCR技术对4个野生草鱼群体(邗江、九江、石首、吴江)和2个选育群体(F1和F2)进行了微卫星序列遗传变异分析。结果显示,6个草鱼群体遗传多样性水平较高,2个选育群体除了平均等位基因数外,其他遗传多样性参数均小于4个野生群体。哈迪—温伯格平衡(Hardy-Weinberg equilibrium)检测显示,在120个群体—位点组合中有62个位点显著偏离哈迪—温伯格平衡,62个群体—位点组合中只有11个组合其近交系数值为负值,其余的51个组合的Fis均为正值。6个草鱼群体AMOVA分析结果显示,3.75%的变异来自于群体间,96.25%的变异来自于群体内,整体的遗传分化指数值为0.038。进一步分析各个群体间Fst,只有石首群体与F1、F2群体之间的Fst大于0.05,处于中等分化,其余群体间分化程度较低,且F2群体与4个野生群体之间Fst比F1群体与4个野生群体之间的Fst大。奈氏标准遗传距离分析结果显示,2个选育群体与野生群体之间的遗传距离大于野生群体之间的遗传距离。基于Dn建立的UPGMA系统发育树得出了相同的结果,即2个选育群体与野生群体之间的亲缘关系比4个野生群体之间的亲缘关系要远。研究表明,经过2个世代选育后,相比4个野生群体,2个选育群体遗传多样性虽有部分下降,但仍处于较高的水平,2个选育群体的遗传结构已发生变化,但其遗传分化程度尚不明显。本研究结果为制定出更加完善有效的选育方案提供了重要参考。     

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.     

基于线粒体控制区序列的光裸方格星虫群体遗传多样性分析

为了解光裸方格星虫的群体遗传结构和种质资源状况,对北部湾海域光裸方格星虫6个地理野生群体(广西北海、湛江、钦州、防城港、海南儋州,以及越南海防)共93个个体的线粒体DNA(mtDNA)控制区序列进行分析。共检测到107个变异位点,定义了85个单倍型,序列对AT有明显的偏倚性,群体总的单倍体多样性(Hd)和核苷酸多样性(Pi)分别为0.998和0.018 89。单倍型邻接聚类树分析6个群体无明显分支,各单倍型分布于单倍型网络中介图中亦没有明显的地理分支。各群体间的遗传分化系数Fst值为–0.018 13~0.028 05,遗传分化极不明显,AMOVA分析显示光裸方格星虫的遗传差异主要来自群体内(99.74%)。中性检验Tajima’s D和Fu’s Fs值均为负值,核苷酸不配对分布图呈明显的单峰形,提示光裸方格星虫群体在历史上曾出现群体扩张,推算出扩张时间为距今171万年前。目前光裸方格星虫仍具有较高的遗传多样性,6个地理群体间无明显遗传分化,存在频繁的基因交流,推测种群在早更新世曾出现群体扩张。     

入侵地和原产地太湖新银鱼群体遗传结构

为了探讨太湖新银鱼快速适应新环境和快速入侵的遗传学机制,本研究利用10个微卫星位点对5个入侵地和2个原产地的群体遗传结构进行了分析。遗传多样性参数结果显示,入侵地滇池、邛海、抚仙湖和三峡库区群体的遗传多样性水平比原产地高,入侵地洱海群体的遗传多样性水平低于原产地太湖而高于巢湖群体,原产地巢湖群体的遗传多样性最低。遗传距离和UPGMA聚类结果表明邛海群体和其他群体的遗传关系最远,太湖与抚仙湖的遗传关系最近。ANOVA显示大多数遗传变异存在于太湖新银鱼群体内(95.78%),群体间的遗传变异为4.22%,固定系数(Fst=0.0422)显著,表明太湖新银鱼群体间存在显著的小尺度遗传分化,两两遗传分化指数也证实了这一点。MVSP主成分分析显示邛海、三峡库区、巢湖和滇池群体有明显的分化。由此推断,高水平的遗传多样性和显著的遗传结构差异性可能是太湖新银鱼成功入侵的重要原因。     

Genetic relatedness and differentiation of hatchery populations of Asian seabass (Lates calcarifer) (Bloch, 1790) broodstock in Thailand inferred from microsatellite genetic markers

After more than 20 years of hatchery production of Asian seabass in Thailand, genetic information is still lacking for effective genetic management and a selective breeding programme. This study aimed to evaluate genetic status of existing hatchery populations and genetic consequences of a selective breeding attempt. We examined genetic relatedness in seven hatchery samples, including a selectively bred population (RACF‐F1), compared with three wild samples using 11 microsatellite loci. Genetic diversity and relatedness values within most hatchery samples, except for RACF‐F1, did not differ from those of wild populations (P > 0.05). RACF‐F1 had the lowest allelic diversity and effective population size (A_r = 6.99; N_e = 7.8) and highest relatedness values (mean r_xy = 0.075–0.204). Pairwise Φ_ST values, principal component analysis and model‐based cluster analyses revealed three genetically distinct hatchery groups: Eastern Thailand (CHN, RACF, NSCF and SKCF), Southern Thailand (NICA) and the Andaman Sea (STCF). Results suggest that exiting domestic populations capture reasonable amount of genetic variation and can be useful for a base population for genetic improvement programmes. In addition, given the rapid increase in relatedness that we observed in one selectively bred population, we recommend using selection methods and hatchery practices that reduce variability in family contribution in the subsequent generations.     

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.     

Domestication of the mud crab <Emphasis Type="Italic">Scylla serrata</Emphasis>

The significant decrease in wild mud crab population highlights the need to manage the resources and domesticate crabs. This paper presents the initial results of the domestication of mud crab Scylla serrata aimed at producing good-quality captive broodstock. The analysis of the genetic structure of the base population was done as a prerequisite for domestication. Adult S. serrata from the northern to southern parts of the Philippines (Cagayan, Camarines, Samar, and Surigao) were obtained for genetic diversity analysis and domestication. Analysis of molecular variance showed that differences in the genetic variability between the four populations were not significant. Moreover, no significant deviation from Hardy–Weinberg Equilibrium was observed in each sample population and even in pooled populations. Body weight was positively correlated with the carapace width. Second spawning occurred 41–46 days after the first spawning and 34 days from second to third spawning. However, there was a decrease in the number of zoea in repeat spawnings. Twenty-four first-generation (F₁) families were produced from the four sites. The duration from spawning of the base population (P₀) to attainment of broodstock size F₁ was 10–14 months. Four second-generation (F₂) families were produced after 11–12 months. Up to the F₂, crabs tested negative for six viruses: white spot syndrome virus, infectious hypodermal and hematopoietic necrosis virus, gill-associated virus, yellow head virus, Taura syndrome virus, and infectious myonecrosis virus. The reproductive performance of P₀ was comparable to the succeeding generations. Several families were obtained from one population in a year. However, due to the cannibalistic behavior of crabs, more space is required for the nursery and grow-out phase. The domestication of S. serrata is the first study done on any mud crab species in the Indo-west Pacific region. The initial results would serve as guide to understand and eliminate the barriers to mud crab domestication. The breeding technology developed from this study will support the production of good-quality seedstock for farming.     

尼罗罗非鱼人工驯养、选育群体遗传多样性及瓶颈效应

人工驯养和选育是家养动物适应性进化的主要动力之一,中国大陆尼罗罗非鱼引进群体经历了长期的人工驯养和选育,是研究鱼类在驯养、选育条件下适应性进化的良好材料。本实验以尼罗罗非鱼1个埃及土著群体为对照组,以中国大陆具有代表性的尼罗罗非鱼5个驯养群体和4个选育群体为实验组,采用12个多态性微卫星位点分析了驯养群体和选育群体的遗传多样性和瓶颈效应。结果显示,土著群体、驯养群体和选育群体平均每个位点的有效等位基因数(AE)分别为5.433、5.113~6.515和3.239~6.734,期望杂合度(HE)分别为0.812、0.796~0.859和0.657~0.858,多态信息含量(PIC)分别为0.768、0.753~0.819和0.601~0.818,近交系数(FIS)分别为0.323、0.166~0.342和0.249~0.314。LSDt检验结果显示,受人工选择的群体(驯养群体、选育群体)与土著群体间遗传多样性水平(AE和HE)无差异,3个驯养群体(EGY群体、WY群体和GD群体)的遗传多样性水平(HE)显著高于1个选育群体(XJF群体)。瓶颈效应分析显示,尼罗罗非鱼土著群体、驯养群体和选育群体在历史上都曾发生过群体缩小的现象。其中,土著群体、2个驯养群体(WY、EGY)、2个选育群体(JNM、XJF)在近期可能经历过遗传瓶颈,其他群体在新的突变和基因流的作用下,群体规模可能已恢复。有效群体大小分析显示,土著群体、驯养群体和选育群体的有效群体数量分别为177、29~117(平均值为57.4)和84~123(平均值为102.8)。本研究结果不仅为尼罗罗非鱼驯养群体的持续利用和选育群体的进一步遗传改良提供了有价值的信息,而且为鱼类在驯养和选育条件下群体遗传结构和种群动态研究提供了新的参考依据。