Genetic diversity in three redclaw crayfish (Cherax quadricarinatus,von Martens) lines developed in culture in China

The redclaw crayfish Cherax quadricarinatus (von Martens) accounts for the entire commercial production of freshwater crayfish in Australia. Two forms have been recognized, an ‘Eastern’ form in northern Queensland and a ‘Western’ form in the Northern Territory and far northern Western Australia. To date, only the Eastern form has been exported overseas for culture (including to China). The genetic structure of three Chinese redclaw crayfish culture lines from three different geographical locations in China (Xiamen in Fujian Province, Guangzhou in Guangdong Province and Chongming in Shanghai) were investigated for their levels and patterns of genetic diversity using microsatellite markers. Twenty‐eight SSR markers were isolated and used to analyse genetic diversity levels in three redclaw crayfish culture lines in China. This study set out to improve the current understanding of the molecular genetic characteristics of imported strains of redclaw crayfish reared in China. Microsatellite analysis revealed moderate allelic and high gene diversity in all three culture lines. Polymorphism information content estimates for polymorphic loci varied between 0.1168 and 0.8040, while pairwise F_ST values among culture lines were moderate (0.0020–0.1244). The highest estimate of divergence was evident between the Xiamen and Guangzhou populations.     

乌苏里江2种细鳞鲑种群遗传多样性及亲缘关系的微卫星分析

尖吻细鳞鲑(Brachymystax lenok)28尾和钝吻细鳞鲑(B.tumensis)26尾,均采自乌苏里江中游。利用9对微卫星引物(10个位点)对其种群遗传多样性进行比较研究,以期从分子水平上探讨2个种间的遗传多样性差异水平及其亲缘关系。结果表明,尖吻细鳞鲑和钝吻细鳞鲑种群的平均杂合度(H)分别为0.323 0和0.339 1,多态信息含量(PIC)分别为0.322 9和0.435 3;在检测到的10个基因位点中有9个位点上的等位基因及其在2种细鳞鲑间的分布存在显著差异,2种细鳞鲑各自拥有大量的特有等位基因,而共有等位基因很少;2种细鳞鲑在4个微卫星位点上的遗传相似度(I)为0,在其他6个具有相同等位基因的位点上的遗传距离(D)为0.011 5~2.575 9,平均为0.939 3。研究表明,2种细鳞鲑在种群遗传多样性上差异显著,亲缘关系很远,物种间可能存在生殖隔离,遗传分化程度远达到种以上的分类水平。本研究从分子水平上验证了对2种细鳞鲑的形态学分类结论,并对2种细鳞鲑的起源及其演化进行了探讨。[中国水产科学,2007,14(1):39-45]     

Genetic diversity and differentiation of Nile tilapia (Oreochromis niloticus) revealed by DNA microsatellites

To assess the genetic diversity of Nile tilapia Oreochromis niloticus, a total of 250 fish from five Egyptian populations were genotyped using six microsatellite markers. Heterozygosity and Wright's F‐statistics (F_IS, F_ST, and F_IT) were calculated to determine the genetic variation within and between these populations. Observed heterozygosities were in the range of 0.4 (Burullus) to 0.96 (Qena), with F_IS values ranging from 0.082 to 0.282. The mean F_ST showed that approximately 96.5% of the genetic variation was within‐population and 3.5% was among populations. Standard genetic distances were used to classify the five populations into two major groups. The deeper lotic river Nile populations of Assuit and Cairo formed one group and the shallow less lotic Delta lakes populations of Manzalla and Burullus formed the second group, with the upstream Nile Qena population being an outgroup. The findings from the current study help understanding of the broad‐scale population structuring of the Nile tilapia (O. niloticus) allowing the population groupings identified to act as potential sources of genetic variation. These populations could be included in future Marker‐Assisted‐Selection programs for economically desired production traits.     

兰州鲇野生群体和人工繁育群体遗传结构的比较研究

采用19对大口鲇(Silurus meriaionalis)微卫星引物对兰州鲇(Silurus lanzhouensis)野生和人工繁育2个不同群体进行微卫星标记的遗传结构分析,结果显示:(1)2个不同种群检测出11个有效微卫星位点,共94个等位基因;各基因座位间除DQ223153与DQ223150,DQ223177与DQ223182,DQ223164与DQ223176存在一定程度连锁(P<0.05),其余连锁关系不显著。(2)2个种群平均有效等位基因数为7.28,平均观测杂合度为0.8873,平均期望杂合度为0.7732,PIC值0.7055,均为高度多态,但人工繁育群体的总扩增位点数和遗传多样性均低于野生群体;(3)两群体间的遗传相似性指数(I)为0.9915,遗传距离(D)为0.0086;各位点F-统计量分析结果表明,群体遗传分化系数(FST)为-0.0392～0.0754,平均值为0.0205;基因流(Nm)为1.6625。综合结果说明,虽然兰州鲇人工繁育群体的遗传多样性降低,但两群体遗传多样性丰富,亲缘关系较近,遗传分化较低,属于同一个种内水平遗传变异。     

Genetic changes during mass selection for growth in Nile tilapia, Oreochromis niloticus (L.), assessed by microsatellites

Two control (C1 or first control generation, and C4 or fourth control generation) and three selected (S1 or first selected generation, S2 or second selected generation, S4 or fourth selected generation) stocks of Chitralada Nile tilapia were analysed for microsatellite variation to determine the effect of size‐specific mass selection on genetic variability. Genetic variation based on five microsatellite loci (UNH123, UNH147, UNH172, UNH222 and UNH216) showed a slightly higher allelic diversity in the selected stocks (7.4–10 alleles) than in the control stocks (6.8–8.8 alleles). Apparent reductions in the mean number of alleles and H_e values were noted in successive generations of both control and selected lines. Significant deviations from Hardy–Weinberg equilibrium because of an excess of homozygotes indicated inbreeding in all control and selected stocks. Although estimated inbreeding levels were not significantly different among selected and control lines based on Welch's t‐tests, the increase in the degree of inbreeding within the selected line was higher (107.9%) than the control line (64.2%) after four generations. The implications of these results on the management and conservation of genetic diversity in improved breeds are discussed, while the importance of monitoring and minimizing inbreeding are likewise emphasized.     

长牡蛎3代人工选育群体的微卫星分析

进行群体选育时,因近交机率增加和有效亲本数的减少,可能导致选育群体的遗传多样性下降,进而引起选育群体的性状衰退。为监测长牡蛎人工选育群体在选育过程中的遗传差异,实验应用微卫星DNA标记对长牡蛎野生和人工3代选育群体及其基础群体的遗传多样性进行了研究。微卫星10个位点在所有群体中均表现出较高的多态性,6个群体的平均等位基因数范围为24.0～29.7个,期望和观测杂合度分别为0.925～0.956和0.724～0.809。与野生群体和基础群体相比,长牡蛎选育3代群体的平均等位基因数和等位基因丰富度略有下降,但杂合度水平未发生明显变化。哈迪—温伯格平衡(HWE)检验结果显示,60个群体—位点组合中47个群体—位点组合显著偏离HWE平衡(P<0.05)。Fis指数均为正值,平均范围0.152～0.233,表明各群体在10个位点上表现为一定程度的杂合子缺失。各群体间Fst值的范围为0.008～0.025,遗传分化程度较弱。结果表明,连续3代的人工选育尚未明显降低长牡蛎群体的遗传多样性,仍可以一定的选择压力对选育群体进行人工选育,从而保证长牡蛎的优良生长性状得到持续提高。     

Genetic diversity and population structure of Sepiella japonica (Mollusca: Cephalopoda: Decapoda) inferred by 16S rDNA variations

In order to describe the genetic diversity and phylogenetic relationship of five populations of cuttlefish (Sepiella japonica) along with China's coasts, partial 16S rDNA (510 bp in length) was amplified from 110 individuals. The five populations of cuttlefish inhabit Yellow Sea, East China Sea and South China Sea. In total, six haplotypes were identified and formed only one clade. Among the six haplotypes, one was shared by all populations, three appeared only in a single population, two appeared in two or three populations. Pair‐wise FST were not proportional to the geographical distances. Haplotype diversity and nucleotide diversity were low, 0.3866 ± 0.067 and 0.00120 ± 0.00081 respectively. Among the five populations, Zhoushan population exhibited the highest genetic diversity which was suggested as the better select of germplasm resources for the reproduction and releasing of S. japonica.     

长江中上游南方鲇遗传多样性的RAPD分析

采用随机扩增多态性DNA(RAPD)技术对取自长江中上游合川、木洞、涪陵、彭水、巫山、宜昌及岳阳7个江段的南方鲇(Silurus meridionalisChen)野生群体的遗传多样性进行了分析,24个随机引物共检测出327个位点(200~2500 bp)。各群体的多态位点比例在28.75%~48.62%之间,基因多样性指数为0.2047~0.3074。遗传距离显示长江中上游南方鲇各群体间有一定遗传分化,其中,木洞和合川群体间遗传距离最小为0.0537),而合川和岳阳群体间的遗传距离最大(0.1205),群体间遗传分化系数为0.1866。聚类分析(UPGMA)显示,7个群体聚为两大类群,合川和木洞亲缘关系最近,首先聚在一起,之后与彭水、涪陵、巫山群体相继聚类形成类群Ⅰ,岳阳和宜昌群体则聚类形成类群Ⅱ。Shannon指数分析表明,长江中上游南方鲇野生群体间种质资源状况良好,遗传多样性水平较高。     

中国沿海脉红螺群体遗传多样性及其遗传结构

我国脉红螺自然资源日趋衰减,为制定相应保护措施需要对脉红螺遗传多样性及遗传结构进行分析研究。利用9个微卫星标记分析了我国沿海9个脉红螺群体的遗传多样性和遗传分化水平。遗传多样性分析结果显示,群体平均等位基因丰度（AR）为8.6~9.5,期望杂合度（HE）为0.705~0.777,观测杂合度（HO）为0.498~0.626。遗传分化分析结果显示,群体间遗传分化指数（FST）范围为0.012 2~0.093 6,各群体间没有显著的遗传分化,遗传距离（DC）为0.212~0.349。本研究结果表明,我国沿海脉红螺群体具有较高的遗传多样性,各群体间没有显著分化。     

Genetic comparison of Iranian and Azeri populations of the oriental bream Abramis brama orientalis (Berg) using microsatellites

The aim of this study was to compare, levels of genetic polymorphism between Iranian and Azeri bream (Abramis brama orientalis) populations using five microsatellite loci. The reduced number of alleles in the Iranian populations compared with the Azeri population led us to infer that the genetic variability of the Iranian stocks has been reduced due to inbreeding and genetic drift. Significant population differentiation (F_st) was observed between the Azeri and the Iranian populations, which could be explained by the higher number of alleles in the Azeri population. Significant deviations from Hardy–Weinberg equilibrium were found at more loci in the Iranian populations than the Azeri population. Beyond the hypothesis of null alleles, the heterozygote deficiency may have arisen due to breeding between related individuals in the Iranian populations. This investigation represents the first approach to the knowledge of the genetic variability of Iranian and Azeri bream populations using microsatellite markers, and the reported results could be of interest for management and conservation programmes of this species in Iran.     

吉富品系尼罗罗非鱼选育过程中遗传变异的微卫星分析

采用微卫星标记技术对选育新品种新吉富罗非鱼（F8－9）群体、基础群体（F0）、以及选育中群体（F6－7）在选育过程中出现的遗传变异进行观察与分析。19个微卫星位点扩增后的等位基因数为3～11个,随引物不同而异,合计得到115个等位基因,大小在70～270 bp之间。五群体平均基因多样性指数分别为,F0：(0.3083±0.1834)、F6：(0.2982±0.1889)、F7：(0.2923±0.1898)、F8：(0.2572±0.1923)、F9：(0.2743±0.1597),表明随着选育的进展,群体呈现出纯化趋势。AMOVA分析表明,在总遗传变异中,93.24%来自选育群体内,仅6.67%来自选育群体间。群体间的校正偏差后相似性系数在 F0与F6、F7之间分别为0.943 5和0.942 2,相对较高;而在F0与F8、F9之间分别为0.933 2和0.930 3, 相对较低。遗传距离在F0与F6、F7之间分别为0.058 1和0.059 5,相对较小;而在F0与F8、F9之间分别为0.069 1和0.072 2,相对较大。配对比较Fst值在F0与F6、F7之间分别为0.037 68和0.064 37,平均 值为0.051 03;而在F0与F8、F9之间分别为0.060 93和0.075 87,平均值为0.068 40,有所提高。这些指标的分析结果表明,经9年9代选育,已在罗非鱼世代间造成程度虽小但却可监测到的遗传分化;同选育群体F6－7相比,\"新吉富罗非鱼\"（F8－9）在遗传上更为稳定。图1表4     

大黄鱼连续4代选育群体遗传多样性与遗传结构的微卫星分析

利用13个多态性微卫星位点分析了大黄鱼官井洋优快01品系F1到F44个选育世代的遗传结构与遗传多样性变化情况。结果显示,随着选育的进行,4个世代群体遗传多样性指标值渐次下降,F1到F413个微卫星位点的平均多态信息含量从0.638下降到0.524,平均等位基因数从5.462下降到4.308,平均观测杂合度从0.779下降到0.532,平均Shannon多样性指数从1.356下降为1.092。F1与其后各代遗传相似系数逐渐减小(从0.7194到0.5813),遗传距离逐渐增加,而相邻世代间的遗传相似性逐步升高,遗传分化指数(FST)渐次变小(F1～F2为0.0619,F2～F3为0.0511,F3～F4则为0.0475)。随着选育的进行,微卫星位点LYC0002和LYC0054等位基因频率有规律地发生变化,推测其可能与选育性状存在遗传上的相关。结果表明,经过连续4代的选育,部分不利基因遭到淘汰,选育群体的遗传基础逐步得到纯化,基因型逐渐趋向纯合、稳定,经进一步的选育可望获得较稳定的品系。     

兰州鲇（CAG）n、（GATA）n微卫星文库构建与鉴定

应用磁珠富集法构建兰州鲇（ Silurus lanzhouensis） CAG重复和GATA重复的微卫星文库,并分析其序列特征。兰州鲇基因组DNA经MseI酶切,选取200～800 bp的片段与生物素标记的探针（CAG）8和（GATA）6杂交,捕获到含有微卫星序列的目的DNA片段连接到pMD19-T载体,转化到大肠杆菌DH5α菌株中构建微卫星富集文库,经PCR检测筛选出阳性克隆进行测序。从126个阳性克隆中随机选取96个进行测序,获得59个微卫星序列（ GenBank登录号： KJ545973～KJ545998, KJ598088～KJ598120）。其中完美型31个（52.54％）、非完美型20个（33.9％）、混合型为8个（13.56％）。根据侧翼序列,成功设计48对引物,选取25对微卫星引物在10个个体进行扩增与多态性筛选,共获得10对多态性引物。结果表明,经优化的磁珠富集法能够高效地获得兰州鲇微卫星标记,这些标记将为兰州鲇种质资源保护、微卫星连锁图谱构建、经济性状的QTL定位及分子标记辅助选育奠定基础。     

不同野生群体与家系养殖翘嘴鳜遗传多样性分析

为探讨长江野生群体、通江湖泊野生群体和家系养殖翘嘴鳜群体的遗传多样性,利用7个高度多态的微卫星标记对5个群体进行遗传分析。结果显示,7个微卫星位点的等位基因数、有效等位基因数平均值分别为29和9。观测杂合度、期望杂合度、平均多态信息量平均值分别为0.853、0.886和0.875。通过平均多态信息量PIC统计发现,遗传多样性从大到小依次为:长江野生群体>梁子湖群体>洞庭湖群体>武汉养殖群体>无锡养殖群体。遗传距离及遗传相似系数分析表明,长江野生群体和无锡养殖群体间的遗传距离最大(0.813),遗传相似系数最小(0.444)。UPGMA聚类分析显示,5个群体可分为2个亚群,其中亚群I包括梁子湖群体、洞庭湖群体和长江野生群体,亚群II包括武汉养殖群体和无锡养殖群体。结果表明翘嘴鳜不同种群间的基因交流受到了一定的地理阻碍,特别是养殖群体与长江野生群体表现较高的遗传分化。     

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.     

Genetic parameter estimates for growth‐related traits of blunt snout bream (Megalobrama amblycephala) using microsatellite‐based pedigree

Blunt snout bream (Megalobrama amblycephala) is an herbivorous freshwater fish species native to China and has been recognized as a main aquaculture species in the Chinese freshwater polyculture system with high economic value. The genetic parameter estimates for important economic traits are needed for its selective breeding. The aim of this study was to estimate the heritabilities for its growth‐related traits and explore the genetic and phenotypic correlations among the traits using microsatellite‐based pedigree approach. Offspring from a mass‐spawning of 92 broodstocks (42 sires and 50 dams) were reared in a communal pond and nine microsatellites were used to identify the parents of each sampled offspring. Of 749 offspring randomly selected, 708 (94.53%) could be assigned directly to a single parental pair, which was used for heritability estimates. Data were analysed using the method of restricted maximum likelihood (REML) using animal model and the results showed that the heritabilities of body weight, body length, total length and body height were 0.65, 0.53, 0.53 and 0.50, respectively. High genetic correlations were found among these four traits. According to these results, selection for growth seems to be feasible in M. amblycephala and the other growth traits will be heightened accordingly with the selection based on body length.     

呼玛河3种茴鱼遗传分化及属内地位的微卫星分析

茴鱼属（Thymallus）是黑龙江水系特有的珍稀濒危冷水性鱼类,本研究利用6对微卫星引物,对采自黑龙江上游呼玛河同域分布的黑龙江茴鱼（T.grubii）、下游黑龙江茴鱼（T.tugarinae）及待定名种（T.sp.）3种茴鱼的遗传多样性进行了比较研究,以探讨其种群遗传结构、分化水平及分类学地位,为制定保护管理策略提供遗传学理论依据。结果显示,黑龙江茴鱼、下游黑龙江茴鱼和待定名种的平均等位基因数（Na）分别为17.3、18.2和15.3,平均杂合度（H）分别为0.9112、0.9190、0.8492,多态信息含量（PIC）为分别为0.8900、0.8980和0.8250,种群中的特有等位基因数分别为34、37和43。黑龙江茴鱼与下游黑龙江茴鱼的遗传距离（Ds）最小（0.9617）,下游黑龙江茴鱼与待定名种最大（1.8427）,3种茴鱼的Fst为0.1575（P〈0.01）,远大于黑龙江茴鱼、下游黑龙江茴鱼的不同地理种群间的遗传分化水平。研究表明,黑龙江上游3种茴鱼具有很高的遗传多样性水平,3个种间产生显著的遗传分化,结合3种茴鱼种间稳定而显著的形态学特征差异及其地理分布现状,支持黑龙江上游呼玛河同域分布的3种茴鱼在属内各为独立种的分类学地位。     

3个仿刺参地理种群遗传变异的微卫星DNA分析

运用微卫星DNA技术对仿刺参烟台群体、威海群体、大连群体的3个野生群各20个个体进行了遗传分析。对9个基因位点进行了扩增,共获得了59个等位基因。评估了9对微卫星引物的多态信息含量(PIC),范围在0.5129~0.8794之间。结果表明:3个野生群体的平均杂合度观测值分别为0.6416、0.6595、0.5824,平均杂合度期望值分别为0.7641、0.7161、0.7364;在Hardy-Weinberg平衡条件下,进行了P检验,发现3个群体均有位点发生了显著偏离;对3个群体进行了配对Fst值计算,发现3个群体之间遗传分化较弱。从变异贡献率来看,95.68%的变异来自个体之间,4.32%的变异来自群体之间。经过聚类分析,发现威海群体和大连群体之间亲缘关系较近,烟台群体与前两群体亲缘关系较远。     

Genetic diversity of cultured and wild populations of the giant freshwater prawn Macrobrachium rosenbergii (de Man, 1879) based on microsatellite analysis

Freshwater prawn (Macrobrachium rosenbergii) culture in the Western Hemisphere is primarily, if not entirely, derived from 36 individual prawns originally introduced to Hawaii from Malaysia in 1965 and 1966. Little information is available regarding genetic variation within and among cultured prawn stocks worldwide. The goal of the current study was to characterize genetic diversity in various prawn populations with emphasis on those cultured in North America. Five microsatellite loci were screened to estimate genetic diversity in two wild (Myanmar and India‐wild) and seven cultured (Hawaii‐1, Hawaii‐2, India‐cultured, Israel, Kentucky, Mississippi and Texas) populations. Average allelic richness ranged from 3.96 (Israel) to 20.45 (Myanmar). Average expected heterozygosity ranged from 0.580 (Israel) to 0.935 (Myanmar). Many of the cultured populations exhibited reduced genetic diversity when compared with the Myanmar and the India‐cultured populations. Significant deficiency in heterozygotes was detected in the India‐cultured, Mississippi and Kentucky populations (overall F_is estimated of 0.053, 0.067 and 0.108 respectively) reflecting moderate levels of inbreeding. Overall estimate of fixation index (F_st = 0.1569) revealed moderately high levels of differentiation among the populations. Outcome of this study provide a baseline assessment of genetic diversity in some available strains that will be useful for the development of breeding programmes.     

呼玛河3种茴鱼遗传分化及属内地位的微卫星分析

茴鱼属(Thymallus)是黑龙江水系特有的珍稀濒危冷水性鱼类,本研究利用6对微卫星引物,对采自黑龙江上游呼玛河同域分布的黑龙江茴鱼(T.grubii)、下游黑龙江茴鱼(T.tugarinae)及待定名种(T.sp.)3种茴鱼的遗传多样性进行了比较研究,以探讨其种群遗传结构、分化水平及分类学地位,为制定保护管理策略提...