Allozyme genetic analysis in hatchery strains and wild blue shrimp, Penaeus (Litopenaeus) stylirostris (Stimpson), from the Gulf of California

We conducted a population genetic analysis of Penaeus stylirostris to describe the genetic variability in two wild samples (Guaymas and Peñasco) and in two cultivated strains (A and B), and to determine the relationship among samples. Seventeen enzymatic systems and general proteins were used to visualize 31 loci. Of the 31 loci, 11 polymorphic loci (35%) were detected in all the organisms, with an average heterozygosity of 0.10 and an average of 1.8 per locus. There were no significant differences among the four samples of observed heterozygosity or percentage of polymorphic loci. According to Hardy–Weinberg, both cultivated strains showed disequilibrium in more loci than the wild samples. Guaymas showed linkage disequilibrium in five pairs of polymorphic loci, Peñasco samples in two pairs, A in seven pairs, and B in nine pairs. Peñasco and both cultivated lineages showed evidence of recent reductions in their effective populations size. Based on the comparisons of F_st values, and the distribution of the allelic frequencies, the cultivated strains were significantly different from the two wild populations. There is evidence that strain B originated from A and the discrepancies between the strains are probably the result of genetic drift and a founder effect.     

Genetic differentiation in hatchery strains and wild white shrimp Penaeus (Litopenaeus) vannamei (Boone, 1931) from northwest Mexico

Ten enzymatic systems were analyzed to determine allozyme genetic differentiation among three hatchery strains (A, B and C) of white shrimp, Penaeus (Litopenaeus) vannamei, commonly used in shrimp farming in northwest Mexico. A wild population from northern Sinaloa was used as a reference. Fifteen loci were detected, nine of which were polymorphic (ACP-1*, ACP-2*, AKP-2*, EST-2*, EST-3*, EST-4*, EST-5*, LAP*, and LDH*). Polymorphism of A, B and C were 53, 53, and 40%. The mean observed heterozygosity per locus was 0.071, 0.093, and 0.050 without any significant difference among them or with respect to the wild population (0.056). In all samples observed heteroxygosity was smaller than expected. The mean number of alleles per locus was 1.8 in A and 1.87 in both B and C. Strain A was the only sample without rare alleles. Only EST-3* and LAP* of strain A were in Hardy-Weinberg equilibrium; the other loci were in disequilibrium in all samples. High inbreeding values and heterozygote deficiencies were detected in all samples. Distribution of allelic frequencies was heterogeneous among the samples (G-test), involving all polymorphic loci and suggesting a genetic differentiation. According to Fst, a moderate genetic differentiation (7.4%) was detected among the samples. Greater differences were between strains A and C. Based on genetic distance, the samples were grouped into two pairs, B–C and A-wild. Strain A is a young strain related to the wild sample, whereas strains B and C have a different geographic origin than the wild sample.     

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。     

Genetic variation of the razor clam Ensis siliqua (Jeffreys, 1875) along the European coast based on random amplified polymorphic DNA markers

Ensis siliqua is regarded as an increasingly valuable fishery resource with potential for commercial aquaculture in many European countries. The genetic variation of this razor clam was analysed by randomly amplified polymorphic DNA (RAPD) in six populations from Spain, Portugal and Ireland. Out of the 40 primers tested, five were chosen to assess genetic variation. A total of 61 RAPD loci were developed ranging in size from 400 to 2000 bp. The percentages of polymorphic loci, the allele effective number and the genetic diversity were comparable among populations, and demonstrated a high level of genetic variability. The values of Nei's genetic distance were small among the Spanish and Portuguese populations (0.051–0.065), and high between these and the Irish populations. Cluster and principal coordinate analyses supported these findings. A mantel test performed between geographic and genetic distance matrices showed a significant correlation (r=0.84, P<0.05), suggesting an isolation by distance process.     

黄海和东海带鱼群体同工酶分析

采用水平淀粉凝胶电泳技术,对采自中国黄海和东海两个自然群体带鱼(Trichiurus haumela)肌肉和肝脏两种组织的9种同工酶(AAT、ALP、G3PDH、IDHP、LAP、LDH、MDH、PGM、SOD)进行了电泳分析,共检测出13个基因座位,其中3个(AAT*、PGM*、SOD*)为多态位点。两个群体的多态位点比例(P0*.99)分别为0.1538和0.0769,平均等位基因的有效数为1.0211和1.0164,平均观测杂合度(Ho)为0.0192和0.0038,平均预期杂合度(He)分别为0.0173和0.0037,群体间遗传距离(D)为0.0015。     

Genetic characterizations of Mactra veneriformis (Bivalve) along the Chinese coast using ISSR-PCR markers

The genetic structure of seven different populations of the surf clam Mactra veneriformes along the coast of China was investigated by Inter-Simple Sequence Repeats (ISSR) fingerprinting of 210 individual clams. Of the 240 ISSR loci tested, 235 (97.9%) were polymorphic. We found a clear tendency for higher F_ST values and lower gene flow levels between the populations with increasing geographical separation. The seven different geographic populations can be divided into three subgroups: Liaoning, Qingdao/Lianyungang, and Ningbo based on the UPGMA dendrogram of Nei's genetic distance. These results indicated that isolation of geographic distance played an important role in population differentiation.     

Allozymic variation and differentiation in naturalized populations of rainbow trout, Oncorhynchus mykiss (Walbaum), from southern Chile

An allozyme survey was conducted in three naturalized populations of rainbow trout, Oncorhynchus mykiss (Walbaum), from rivers in two Andean lakes of southern Chile. The analysis was based on 32 presumed loci, 10 of which were variable, and revealed that the mean percentage of polymorphic loci (P= 21.8%; range = 15.6-28.1%), the mean number of alleles per locus (n=1.26; range = 1.2-1.3) and the mean observed heterozygosity (H_O=7.0%; range = 6.0-8.2%) are high, although within the range reported for rainbow trout. Although the mean genetic distance between the samples was low (D = 0.003), and only 5.2% of the total observed genetic variation can be accounted by interpopulation differences (F_ST= 0.052), eight out of 10 polymorphic loci exhibited significant allelic heterogeneity. Some loci, notably EST-1*, IDHP-3,4*, MEP-1* and PGM-1*, showed considerable discriminatory capacity among the samples. Rainbow trout has a longer history in Chilean waters since salmonid introduction started earlier this century. Under current intensive farming practices, the preliminary information provided in the present paper is likely to be helpful in setting up a baseline for further characterization and/or monitoring of naturalized populations.     

基于SSR和RSAP的龙须菜作图群体多态性标记 开发与亲本间遗传距离的估计与比较

遗传连锁图谱的构建是实现分子标记辅助育种与数量性状定位的必要手段,为构建龙须菜(Gracilariopsis lemaneiformis)遗传连锁图谱提供多态性分子标记,本实验共采用400对SSR引物和91对RSAP引物对组合,对龙须菜两个F1代配子体作图群体的亲本进行了多态性标记筛选,并对4个亲本间的遗传距离进行了评估.得到的SSR多态性标记在GL-F3/GL-M8间为15个,GL-F6/GL-M9间为8个;多态性RSAP标记在GL-F3/GL-M8间为88个;GL-F6/GL-M9间为93个.分析了基于SSR、RSAP技术的两对亲本间的遗传距离,SSR结果显示GL-F3/GL-M8间遗传距离为0.071 2,GL-F6/GL-M9间的遗传距离为0.043 6;RSAP结果显示GL-F3/GL-M8间遗传距离为0.228 4,GL-F6/GL-M9间的遗传距离为0.253 7.结果表明,在引物组合平均覆盖的位点和引物组合平均产生的多态性方面,RSAP均远高于SSR,在GL-F3/GL-M8间RSAP分别是SSR的4.48与12.13倍,在GL-F6/GL-M9间RSAP分别是SSR的5.81与25.5倍,RSAP标记更适合应用于龙须菜的遗传距离估计和资源的遗传多样性研究.本研究揭示出作图群体亲本间遗传多样性特征,SSR与RSAP分子标记的开发为龙须菜遗传连锁图谱的构建奠定了基础.     

Genetic analysis of Rutilus rutilus caspicus (Jakowlew 1870) populations in Iran by microsatellite markers

The Caspian roach (Rutilus rutilus caspicus) is regarded as one of the most important commercial species in the Caspian Sea. Despite the commercial and conservation importance of the species, information on genetic relationships and diversities at the molecular level in Rutilus rutilus caspicus is scarce. Six microsatellite loci were analysed to study the genetic variation in two major populations of Caspian roach in the Iranian waters of the Caspian Sea: the Anzali Wetland and Gorgan Bay. The F_st value between populations was 0.07, suggesting that the genetic differentiation was significant (P<0.01). Both the studied populations deviated from Hardy–Weinberg equilibrium proportions at a number of loci, mostly due to the deficiency of heterozygosities. The genetic distance between populations was 0.29, which indicates that the genetic difference among the studied populations is pronounced. The differences between both populations were not statistically significant (P>0.05), neither for average number of alleles per locus nor for observed heterozygosities. The data generated in this study provide useful information on the genetic variation and differentiation in populations of Caspian roach.     

Genetic variation and population structure of the caramote prawn Penaeus kerathurus (Forskäl) from the eastern and western Mediterranean coasts in Tunisia

Ten populations of Penaeus kerathurus (Forskäl), a prawn of high commercial value, were sampled from the eastern and western Mediterranean coastal waters of Tunisia and screened electrophoretically for genetic variation at 13 allozyme loci. Four among the six polymorphic loci were out of Hardy–Weinberg equilibrium (H–WE) in at least one population. In the same way, the multilocus test showed deviation from H–WE in six populations. These populations showed heterozygote deficiency whereas the average heterozygosity for the four remaining ones is quite similar to the expected levels. Genetic variability was low. The number of alleles per locus ranged from 1.2 to 1.5 (average=1.3), and the observed heterozygosity varied between 0.010 and 0.048 (average=0.021). Significant population differentiation (F_ST=0.076, P<0.05) in the total data set reflected the differentiation of the two populations, which were at the margins of the range sampled, from all the others (Pairwise F_ST values ranged from 0.035 to 0.208). Although there was no significant differentiation among the other populations (pairwise F_ST values ranged from ?0.006 to 0.201, P>0.05). Our data suggest a population structure consistent with separation by Mediterranean Sea basins that might reflect different local biogeographical zones.     

Genetic variability of cultured populations of the Pacific abalone (Haliotis discus hannai Ino) in China based on microsatellites

The genetic diversity of cultured populations of the Pacific abalone (Haliotis discus hannai Ino) from northern China was analysed using seven microsatellite markers. The microsatellite loci were polymorphic for all the populations, with an average of 8.7 alleles per locus (range 8.0–9.4). The mean observed and expected heterozygosities were 0.547 (range 0.500–0.596) and 0.774 (range 0.754–0.787) respectively. The allelic diversity in terms of number of alleles per locus was considerably lower than that previously found in wild populations (range 21.8–23.0), indicating that bottleneck effects occurred when each population was founded. Significant genetic differentiation among the five cultured populations was shown using F_st and R_st values, and pairwise comparison based on allelic distribution. A neighbour‐joining analysis of the genetic distance did not show a consistent relationship between the geographic and the genetic distances, suggesting the existence of exchanges of breeds and eggs between the hatcheries. The results obtained in this study are useful for a number of areas of interest for fisheries management and the aquaculture industry, especially with regard to breeding programmes.     

白梭吻鲈3个群体的遗传结构比较

为了加强良种选育工作,切实做好亲本遗传管理,防止近交衰退的发生,利用20个微卫星分子标记对白梭吻鲈(Sander lucioperca)新疆野生群体及山东和苏州2个养殖群体的遗传结构进行检测。结果表明,20个微卫星标记中18个有扩增产物,14个呈现多态性;每个位点的等位基因数为2~6个,平均等位基因数为3.6个,3个群体的平均等位基因数为2.57~3.36,平均观测杂合度为0.5085~0.5621,平均多态性信息含量为0.3931~0.4764,表明3个白梭吻鲈群体的遗传多样性处于中等偏低水平,遗传多样性大小为:新疆群体苏州群体山东群体。群体的Fst为0.1798,表明群体间有一定的遗传分化。在白梭吻鲈人工繁殖与养殖过程中,必须加强亲本遗传结构监测并维持一定数量的亲本规模,以利于其产业的持续发展。     

Genetic population structure of goldsinny wrasse, Ctenolabrus rupestris (L.), in Norway: implications for future management of parasite cleaners in the salmon farming industry

The application of goldsinny wrasse, Ctenolabrus rupestris (L.), as parasite cleaners in the salmon farming industry has led to transport of live wrasse between regions in Norway. Owing to the operation of salmon farms, some of the transferred wrasse escape from the salmon net pens and can therefore mix and potentially interbreed with the local populations. The genetic effects of such interbreeding are unknown and information about the genetic population structure of this species in Norwegian inshore waters is needed to evaluate the potential effects. A genetic study based on polymorphic tissue enzymes was therefore carried out. Four of the 35 protein coding loci screened were polymorphic at the 99% level, and only two at the 95% level. The allele frequencies of the IDDH-1*; PGM-1*; GPI-1* and GPI-2* loci were compared in wrasse samples (1442 specimens) collected at 12 localities. Significant differences were found for PGM-1* among the samples collected at two inner fjord localities. Significant differences at the IDDH-1*, PGM-1* and GPI-2* loci were found when data from a sample of transferred wrasse caught in southern Norway were compared with local wrasse in the recipient areas.     

基于线粒体Cyt b基因的黄鳍马面鲀种群分析

采用线粒体细胞色素b(cytochrome b,Cyt b)基因片段为遗传标记,分析了南海北部陆架和南沙西南部陆架海域5个黄鳍马面鲀群体的遗传结构,以判定南海北部不同海域之间及南海北部与南沙西南部陆架海域黄鳍马面鲀的种群归属。结果表明,在156个个体的779 bp Cyt b同源序列中共检测到56个变异位点和58种单倍型,5个群体间遗传距离为0.002 95~0.004 15,遗传分化性呈现出高单倍型多样性(0.820 1~0.980 4)和低核苷酸多样性(0.002 62~0.004 69)的特点;分子方差分析和遗传分化指数显示,黄鳍马面鲀的遗传变异来自群体内个体间,群体间无显著遗传分化;单倍型网络结构图和群体系统发育树结构均未出现明显的以地方群体为单位的家系式分支或者聚簇。比较5个不同地理群体间的遗传发育关系并结合种属界定标准判定,南海北部和南沙西南部陆架5个黄鳍马面鲀群体属于同一个种群。     

Identification and characterization of SSRs in <Emphasis Type="Italic">Clarias batrachus</Emphasis> and their application in population study

Magur Clarias batrachus is an indigenous catfish, commonly found in India, Nepal, Bhutan and Bangladesh. Presently, the populations of magur have declined rapidly in their natural habitat mainly due to overexploitation and habitat degradation. Understanding the population genetic structure of the species has significance in improvement of stocks and in conservation of the species. In the present study, simple sequence repeat (SSR) markers were used to differentiate the populations of magur, collected from three geographic locations. For this, a total of 31,814 SSRs were mined from the de novo assembled pooled of whole genome sequence data of C. batrachus. A bioinformatics pipeline with stringent criteria was applied to analyze the data which resulted in selection of 30,142 microsatellite loci falling in the intergenic region. Out of the 25 loci selected for primer development, 16 loci were successfully amplified and 9 loci were found to be polymorphic in this species. The average observed as well as expected heterozygosity in the loci across different stocks varied from 0.652 to 0.688 and 0.864 to 0.873, respectively. These three populations were further segregated into two clusters based on the NJ genetic distance. The Lucknow population formed one cluster, while the Bhubaneswar and Kolkata populations constituted another cluster. A comparable finding was also deduced from the STRUCTURE analyses. The results revealed significant variation among the populations of C. batrachus under study.     

Genetic differentiation in hatchery strains and wild white shrimp Penaeus (Litopenaeus) vannamei (Boone, 1931) from northwest Mexico

Ten enzymatic systems were analyzed to determine allozyme genetic differentiation among three hatchery strains (A, B and C) of white shrimp, Penaeus (Litopenaeus) vannamei, commonly used in shrimp farming in northwest Mexico. A wild population from northern Sinaloa was used as a reference. Fifteen loci were detected, nine of which were polymorphic (ACP-1*, ACP-2*, AKP-2*, EST-2*, EST-3*, EST-4*, EST-5*, LAP*, and LDH*). Polymorphism of A, B and C were 53, 53, and 40%. The mean observed heterozygosity per locus was 0.071, 0.093, and 0.050 without any significant difference among them or with respect to the wild population (0.056). In all samples observed heteroxygosity was smaller than expected. The mean number of alleles per locus was 1.8 in A and 1.87 in both B and C. Strain A was the only sample without rare alleles. Only EST-3* and LAP* of strain A were in Hardy-Weinberg equilibrium; the other loci were in disequilibrium in all samples. High inbreeding values and heterozygote deficiencies were detected in all samples. Distribution of allelic frequencies was heterogeneous among the samples (G-test), involving all polymorphic loci and suggesting a genetic differentiation. According to Fst, a moderate genetic differentiation (7.4%) was detected among the samples. Greater differences were between strains A and C. Based on genetic distance, the samples were grouped into two pairs, B-C and A-wild. Strain A is a young strain related to the wild sample, whereas strains B and C have a different geographic origin than the wild sample.     

不同地理群体乌鳢线粒体DNA控制区结构分析及遗传多样性

为研究乌鳢群体的遗传多样性和控制区结构,实验采用PCR和DNA测序技术对其线粒体DNA控制区序列进行比较。结果显示,用于分析的线粒体DNA控制区全长序列为905~908 bp。104个序列中共发现了37个多态位点,定义了27种单倍型。同时对控制区结构进行分析,识别了其终止序列区(ETAS)、中央保守区(CD)和保守序列区(CSB)的关键序列。3个地理群体的单倍型多样性(Hd)、核苷酸多样性(Pi)和平均核苷酸差异数(k)分别为0.875、0.003 27和2.939。群体间的平均Kimura双参数遗传距离(Kimura 2-parameter distance,K 2-P)、遗传分化指数(Fst)、基因交流值(Nm)和分子方差分析(AMOVA)均表明,3个乌鳢群体具有较高的遗传分化,白洋淀群体和平原县群体间存在一定的基因交流。     

漠斑牙鲆微卫星标记筛选及美国群体遗传结构分析

采用磁珠富集法筛选适合漠斑牙鲆遗传多样性分析的微卫星分子标记。筛选共获得43条序列,其中完美型26个,占60.5%;非完美型14个,占32.6%;复合型3个,占6.9%。选取其中14对特异性好且扩增效率高的微卫星引物,对采自美国北卡罗来纳州沿海的漠斑牙鲆野生群体和养殖群体进行遗传多样性及遗传结构比较分析。研究结果表明,12对引物的扩增产物具有多态性,其中7个位点为高度多态(PIC>0.5)。两个群体中共检测到90个等位基因。12个多态性微卫星位点在两个群体中的平均观察杂合度(Ho)和期望杂合度(He)分别为0.36和0.57。9个位点在整个群体中呈现出不同程度的偏离遗传平衡(P<0.05),且偏离平衡的位点均表现为杂合子缺失(Fis>0)。野生群体和养殖群体间的遗传距离为0.1115,群体间的遗传分化微弱(Fst=0.0438)。     

Isolation and characterization of 16 polymorphic microsatellite markers from Nibea albiflora

Nibea albiflora is a commercially important fish species in China. Herein we report 16 novel polymorphic microsatellite markers in Nibea albiflora by using the 5′ anchored polymerase chain reaction (PCR) technique. The characteristics of these loci were estimated by using a sample of 30 individuals. A total of 79 alleles were detected with an average of 4.9 alleles per locus. The number of alleles per locus ranged from three to nine. The polymorphism information content (PIC) values for the 16 microsatellite loci ranged from 0.3131 to 0.7910. The observed and expected heterozygosity per locus ranged from 0.2333 to 1.000 and from 0.3452 to 0.8421, with an average of 0.7248 and 0.6592, respectively. Four loci significantly deviated from Hardy–Weinberg equilibrium after Bonferroni correction (P < 0.0031), and no significant linkage disequilibrium between pairs of loci was found. This study provides useful information for studies on genetic diversity and structure, construction of genetic linkage maps of N. albiflora, and effective management of this fish resource.     

不同鳊鲂鱼类群体微卫星DNA指纹图谱的构建和遗传结构分析

为对不同鳊鲂鱼类进行群体鉴定和遗传多样性分析,从60对微卫星标记中筛选出18对多态性高的引物,构建了6个鳊鲂鱼类群体的微卫星DNA指纹图谱。结果显示,东江三角鲂、钱塘江三角鲂、厚颌鲂、广东鲂、团头鲂和长春鳊6群体的平均等位基因数(N a)分别为5.17、6.11、3.50、6.56、5.22、5.22,平均期望杂合度(H e)分别为0.634 2、0.720 4、0.546 2、0.681 2、0.675 2、0.559 7,平均多态信息含量(PIC)分别为0.575 6、0.666 9、0.472 0、0.630 6、0.606 4、0.517 0,表明钱塘江三角鲂的遗传多样性最高,厚颌鲂的遗传多样性最低;聚类分析表明,钱塘江三角鲂和团头鲂首先聚为一支,遗传距离较近,为0.560 6;厚颌鲂与长春鳊的遗传距离最远,为1.759 2。引物Mam03和EST37产生的特异条带可将鲂属和鳊属鱼类区分,鉴定出鳊属鱼类长春鳊;引物TTF3、EST37、TTF2/TTF10、EST66依次组合可区分出鲂属东江三角鲂、厚颌鲂和广东鲂这3个群体。研究结果为我国鳊鲂鱼类种质资源保存、种群鉴定和良种选育奠定了基础。