菲律宾蛤仔大连群体不同世代的遗传多样性

采用12对有效微卫星引物对大连群体菲律宾蛤仔连续4个选育世代(F₁、F₂、F₃、F₄)的144个个体进行了遗传多样性分析。结果表明:共获121个等位基因,每个位点的等位基因数在2-6个不等,其大小在101-273 bp之间;各个世代平均等位基因数在3.75-4.58,平均观测杂合度在0.3391-0.3860之间。从F-检验结果上看,所有世代内有2个位点遗传分化较弱,8个位点遗传分化中等,2个位点遗传分化较大;配对比较F_st值(0.05-0.15)表明4个世代群体间遗传分化程度中等。F_is值表明有2个世代位点杂合度处于过剩状态;但对连续4个世代而言,每个世代均表现出一定程度的杂合子缺失。随着世代连续选育的进行,Nei氏遗传相似性逐渐减小(0.8203-0.8107-0.8031);遗传距离逐渐增大(0.1918-0.2099-0.2129);不同世代群体间遗传相似性系数为0.7873-0.8685,遗传距离为0.141-0.2391。4个世代平均PIC值为0.5055,表明选育后代遗传多样性较好,还有较大的选育潜力,可以继续进行上选。     

长江中上游两个鲢群体遗传变异的微卫星分析

对长江中上游2个鲢群体使用39个微卫星标记进行了遗传多样性分析, 计算并统计了平均观测等位基因数、平均有效等位基因数、多态信息含量、遗传杂合度、Hardy-Weinberg平衡偏离指数、遗传相似系数、遗传距离等遗传参数。结果表明: 万州鲢和监利鲢群体所检测微卫星位点的平均观测等位基因数分别为6.128和4.974; 平均有效等位基因数分别为4.107和3.395; 多态位点百分率分别为100和94.87; 39个微卫星标记共有等位基因259个, 173个等位基因为两群体所共有; 多态微卫星位点的PIC在0.077~0.865之间变动,平均为0.617; 两群体所检测位点平均观测杂合度为0.834和0.775, 平均期望杂合度为0.713和0.623; 两个群体间的遗传相似系数为0.618, 群体间的遗传距离为0.482。结果显示长江中上游两个鲢群体间存在显著遗传分化, 应隶属于不同的种群。     

利用17个微卫星标记分析鳙鱼的遗传多样性

选用本实验室克隆的17个鳙鱼微卫星分子标记分析四川泸州和江西鄱阳湖的两个种群鳙鱼的遗传多样性及种质特性,计算和统计了杂合度、多态信息含量（PIC）、有效等位基因数、等位基因频率、遗传距离、遗传相似系数、Hardy-Weinberg平衡偏离指数等方面内容。结果表明：选择使用17个微卫星标记,其中有4个为单态标记,13个为多态标记。江西和四川鳙鱼群体每个微卫星位点的平均等位基因数分别为3.325及3.882,平均有效等位基因数分别为3.531及2.676,多态位点百分率分别为82.4及70.5, 17个微卫星标记共有等位基因71个,多态微卫星位点的PIC在0.114~0.960之间变动,平均为0.417 ,两群体位点平均观测杂合度为0.385和0.452,平均期望杂合度为0.360和0.422,两个群体间的遗传相似系数为0.897,群体间的遗传距离为0.109。     

不同年龄段大连群体菲律宾蛤仔EST-SSR多样性

为查明年龄结构对菲律宾蛤仔同一群体内遗传多样的影响,采用14个SSR分子标记对大连石河不同年龄段的野生蛤仔进行了检测。结果表明:不同年龄段(1龄-Age1、2龄-Age2、3龄-Age3)蛤仔均维持着较高的遗传多样性。根据POPGENE 1.31和SPSS16.0统计分析显示,位点Rp-11、Rp-12、Rp-19对3个年龄段蛤仔的等位基因数差异极显著(P<0.01);位点Rp-20、Rp-24、Rp-27、Rp-30对其差异显著(P<0.05);剩余7个位点表现为差异不显著(P>0.05)。在平均水平上,每位点等位基因数目Na为4.3095,有效等位基因数目Ne为2.3729,多态位点百分数P(%)为14。观察杂合度和期望杂合度都比较高,观察杂合度平均为Ho=0.2335,期望杂合度平均为He=0.5140。而且,Ne和He随年龄的变化表现出Age2>Age3>Age1的趋势。各年龄段蛤仔——Age1、Age2、Age3的平均观察杂合度(Ho)和平均期望杂合度(He)分别为0.2357、0.2546、0.2159和0.4951、0.5286、0.5184。Age2的遗传多样性指数高于Age1及Age3,遗传分化相对较低。其中,Age1与Age3蛤仔遗传距离最小,D为0.0195,即变异很小;而Age1与Age2遗传距离较大,D为0.0437,变化范围不大(0.0195—0.0437)。从遗传一致度的数值上看了3个年龄段蛤仔的遗传相似程度很大,平均为0.9655。Age1与Age3遗传相似程度高达0.9807,而Age1与Age2相似程度较小为0.9572。说明不同年龄段蛤仔相似程度非常高。根据不同年龄段蛤仔的遗传距离,采用UPGMA平均聚类方法对其进行聚类可知,Age3与Age1蛤仔间遗传距离较小,与Age2蛤仔差异较大。通过对等位基因频率进行卡方检验发现,随着年龄结构的变化,部分基因基因频率减小;同时随着年龄的增长,有部分等位基因得到了纯化。大连群体蛤仔总的遗传分化较低,其遗传分化指数Fst为0.0248(Fst<0.05),遗传分化系数为0.02,说明总的遗传变异中有2%来自于不同年龄段的蛤仔之间。遗传距离和遗传一致度均值分别为0.035和0.9655,基因流(Nm=9.8238)相对流畅,进一步表明年龄结构对蛤仔种群内遗传分化的影响较小。     

红色原鸡群体遗传多样性

为了阐明红色原鸡的群体遗传结构,以对其有效保护提供遗传学依据,采用33个微卫星标记对其群体中56个个体进行了PCR-聚丙烯酰胺多态性电泳检测。33个微卫星座位共检测到140个等位基因,所有座位都呈现出多态性,每个座位的等位基因数在2～8个之间,平均每个座位等位基因数4.24个,有效等位基因数3.30个。根据等位基因频率,计算出的群体表观杂合度、期望杂合度及多态信息含量分别为0.7980、0.6506和0.5948。结果表明,红色原鸡群体遗传多样性较丰富。     

湘江野鲤养殖群体和自然群体遗传多样性的微卫星分析

采用微卫星技术,用17对微卫星引物对湘江野鲤养殖群体和自然群体的的遗传多样性进行分析.结果表明:有15对引物扩增出清晰的条带,其中13对引物在群体间呈现多态性;2个群体中,13对多态性引物分别扩增等位基因2～12个,共90个,其中35个等位基因为2群体共有,55个等位基因具有群体特异性,引物平均等位基因数为6.92个,等位基因频率为0.0667～0.8333;养殖群体和自然群体的平均遗传杂合度和平均多态信息含量分别为0.5688、0.5152,0.5860、0.5347;2个群体间遗传相似性指数为0.6762,遗传距离为0.3238,表明湘江野鲤养殖和自然群体遗传多样性均较为丰富,2个群体间遗传变异程度较高.     

基于转录组平台的蛤仔微卫星标记筛选

以菲律宾蛤仔转录组测序所得拼接序列为基础,采用MISA软件进行微卫星分析,对其中的145个微卫星位点进行引物设计,得到具有清晰扩增条带的微卫星位点58个。对大连庄河野生蛤仔群体的扩增结果表明,18个位点显示单态性,40个位点表现为多态性。该群体40个多态性微卫星位点得到的等位基因数在2—6之间,平均等位基因数为3.4250±0.9718,观测杂合度和期望杂合度分别在0.0000—1.0000和0.0615—0.7996之间,平均值分别为0.2727±0.2272和0.4739±0.1902,群体平均Nei指数为0.4664±0.1872。多态信息含量(PIC)在0.0586—0.7529之间,平均值为0.4148±0.1707,其中16个微卫星位点的PIC值大于0.5,为高度多态性,15个位点0.25PIC0.5,为中度多态性,其余9个为低度多态性。经Sequential Bonferroni校正的Hardy-Weinberg平衡检验,有10个位点尚未偏离平衡。基于转录组平台筛选微卫星标记的方法,在很大程度上推动了DNA分子标记的开发。研究开发的微卫星标记可用于蛤仔群体遗传学、遗传连锁图谱构建及其他相关研究,为蛤仔分子标记辅助育种及群体种质保护等工作提供技术支持。     

草鱼种群SSR分析中样本量及标记数量对遗传多度的影响

利用45对微卫星分子标记（SSR）,以草鱼(Ctenopharyngodon idellus)自然群体为实验材料,探讨野生群体遗传多样性研究中所需的最适样本量与标记量。实验设置6个样本量梯度,9个标记量梯度。对等位基因数（Na）、有效等位基因数（Ne）、观察杂合度（Ho）、期望杂合度（He）等遗传多样性指标的变化趋势进行统计分析。结果表明,样本量、微卫星标记的数量和多态性水平对群体遗传多样性均有较大的影响,其中等位基因数与样本量大小呈显著正相关,而杂合度随标记量的增多而剧烈波动。当取样量大于40,标记量大于25时,各遗传参数值趋于稳定。因此,在应用微卫星标记对水产动物自然群体的遗传学研究中,要根据所研究种类的特点,尽可能采样40尾以上,采用25个以上标记,避免由人为选择的偏差对群体遗传多样性水平的正确评估所造成的影响。同时根据上述研究结果,对陕西草鱼自然群体进行了遗传多样性的评估,结果显示该群体平均等位基因数（MNA）、平均有效等位基因数、平均观测杂合度、平均期望杂合度分别为7.26、4.21、0.73、0.68,认为该群体具有较高的遗传多样性。     

鳜原种群体和养殖群体遗传多样性的微卫星分析

本研究利用20对微卫星引物对鳜(Siniperca chuatsi)原种群体和养殖群体进行遗传多样性分析。结果表明,在鳜原种群体中检测到多态性位点14个,养殖群体11个。在两个群体中共检测到等位基因数96个,其中原种群体检测到等位基因数53个,每个位点的等位基因数在1～7之间,平均有效等位基因数为2.7390;养殖群体检测到等位基因数43个,每个位点的等位基因数在1～6之间,平均有效等位基因数为2.1284。原种群体的平均观察杂合度0.5708,Nei氏期望杂合度0.5295,平均多态信息含量PIC0.5353;养殖群体的平均观察杂合度0.3839,Nei氏期望杂合度0.4011,平均多态信息含量PIC0.5043。因此,与养殖群体相比,鳜原种群体仍有丰富的遗传多样性。本研究可为鳜种质资源的保护、监测和遗传育种提供分子水平上的数据。     

三个野生群体日本囊对虾遗传多样性的SSR分析

为了解野生种群日本囊对虾遗传分化和改良遗传育种,用SSR技术对福建厦门(XM)、广东湛江(ZJ)、广西北海(BH)3个地区野生日本囊对虾进行遗传多样性的研究。采用了10对微卫星引物对3个野生种群进行分析,10个微卫星位点在3个种群中均表现为高度的多态性,每个位点平均检测到3.87个等位基因;平均多态信息含量为0.5893;3个群体的观测杂合度分别为0.6243、0.5704、0.4661,全部群体观测杂合度平均为0.5536;期望杂合度分别为0.7193、0.6189、0.6226,全部群体平均期望杂合度为0.6536。这说明3个野生种群在10个微卫星位点上均具有丰富的遗传多样性。基于Nei's遗传距离的聚类分析显示厦门群体和湛江群体的遗传距离较近。     

牙鲆微卫星标记的筛选及群体遗传结构分析

采用生物素-磁珠吸附微卫星与质粒二次检测相结合的方法克隆牙鲆微卫星,并利用所得到的引物分析野生群体遗传结构.共获得2805个阳性克隆,测序得到3120个含微卫星的序列.其中完美型、非完美型和混合型分别占57.97%、7.25%和34.78%.用Primer 3.0软件设计牙鲆微卫星引物,从中筛选出具稳定多态性的30对,分析中国黄海、渤海海域4个不同海区牙鲆野生群体(大连、北戴河、丹东、青岛)遗传结构.结果显示:有效等位基因数为3.93～9.94,平均为6.95;观测杂合度为0.532～0.895,平均为0.753;期望杂合度为0.635～0.902,平均为0.820;Hardy-Weinberg平衡指数(d)的变化范围为-0.247～0.512,其中7个位点表现为杂合子过剩(d>0),其余位点表现为杂合子缺失(d>0).聚类分析显示:4个群体聚为两类,大连与丹东聚为一类,北戴河与青岛聚为一类.     

Microsatellite variability and heterozygote excess in Elymus trachycaulus populations from British Columbia in Canada

Microsatellite markers were used to assess the genetic diversity and population structure in four populations of Elymus trachycaulus from British Columbia and one population of Elymus alaskanus from Northwest Territories. Fourteen microsatellite loci were used in this study. Our results indicated that E. trachycaulus is highly polymorphic, with an average percentage of polymorphic loci of 96.5% over the four populations. Average expected heterozygosity values (H_E or gene diversity) varied from 0.418 to 0.585 with a mean of 0.497. Most of the genetic variation was found within populations (85%) and the differentiation among populations was found to be 15% (F_st = 0.15). Interpopulation genetic distances corresponded well with the geographic distance between the population sites of origin, as well as morphological characteristics. Tests for Hardy–Weinberg equilibrium (HWE) for all loci and all populations revealed that all loci significantly differ from HWE. Subsequent analysis indicated that departure from HWE at some loci was due to an excess of heterozygotes. Possible explanations for heterozygote excess are discussed. The most likely reason for observed heterozygote excess could be due to the polyploidy nature of the species.     

Genetic Polymorphism of Microsatellite DNA in Two Populations of Northern Sheatfish (Silurus soldatovi)

In this article, population variations and genetic structures of two populations of northern sheatfish (Silurus soldatovi) were analyzed using 24 microsatellite loci enriched from southern catfish (S. meriaionalis Chen) by magnetic beads. Gene frequency (P), observed heterozygosity (Ho), expected heterozygosity (He), polymorphism information contents (PIC), and number of effective alleles (Ne) were determined. One population was wild, ripe individuals collected from Heilongjiang River (HNS); the other was cultured fry collected from Songhuajiang River (SNS). The Hardy-Weinberg equilibrium (HWE) was tested by the genetic departure index (d). The coefficient of gene differentiation G_ST and Φ_ST by AMOVA (Analysis of Molecular Variety) was imputed using Arlequin software in this study. In addition, a phylogenetic tree was constructed by UPGMA method based on the pairwise Nei's standard distances using PHYLIP. A total of 1 357 fragments with sizes ranging between 102 bp and 385 bp were acquired by PCR amplifications. The average number of alleles of the two populations was 8.875. Results indicated that these microsatellite loci were highly polymorphic and could be used as genetic markers. The mean values of the parameters P, Ho, He, PIC, and Ne were 0.165, 0.435, 0.758, 0.742, and 5.019 for HNS and 0.147, 0.299, 0.847, 0.764, and 5.944 for SNS, respectively. Although there were differences, there were no significant differentiations except for the locus HLJcf37. These populations to a certain extent deviated from HWE, such as excessive and deficient heterozygote numbers. The value of G_ST was 0.078 and above 98% of the variation were differences among individuals within the population, so the variation between populations was insignificant. Cluster analysis also showed that the relationships among individuals were very close. In conclusion, the microsatellite markers that were developed through this study are useful for genetic analysis and the genetic culture that was proposed in this study has no significant impact on S. soldatovi.     

元江鲤种群遗传多样性

选择12对微卫星标记检测了于2011年采集自元江(红河上游中国江段)5个样点192尾鲤的群体遗传多样性.共检测到201个等位基因,每个位点等位基因2-27个.各群体各位点平均等位基因(NA)12.25-14.67个,平均有效等位基因(NE)8.28-9.73个,平均观察杂合度(Ho)o.7765-0.8037,平均期望杂合度(HE)0.7761-0.8080,平均多态信息含量(PIC)0.7534-0.7843.元江鲤种群192个个体各位点NA、NE、Ho、HE、PIC分别为16.50、11.26、0.7927、0.8049、0.7966,种群遗传多样性水平高.元江鲤群体之间遗传分化小,可作为一个种群管理单元进行管理.增殖放流要防止遗传多样性丧失.     

Isolation and characterization of ten polymorphic microsatellite loci in Ixodes arboricola, and cross-amplification in three other Ixodes species

We characterized ten polymorphic microsatellite loci from the tree-hole tick, Ixodes arboricola. Loci were screened in 11–18 individuals from three Belgian populations and five to ten alleles were found at each locus. Seven loci did not show deviations from Hardy–Weinberg equilibrium conditions and there were no indications for null alleles at these loci. The three other loci showed significant heterozygote deficiencies in at least one population, and a high potential for the occurrence of null alleles. We observed no effect of potential host DNA on the scoring of the microsatellites. Cross-amplification of the microsatellites was tested in eight specimens of three congeneric species: I. ricinus, I. hexagonus and I. frontalis. Depending on the species, six or seven of the loci were amplified in ≥4 of the 8 specimens and were polymorphic in each of these species (except for Ixaf 11 in I. frontalis and I. ricinus). These loci thus provide a tool for population genetic analysis of I. arboricola. The suitability of these markers needs to be further investigated in its congeners.     

Genetic variability of dromedary camel populations based on microsatellite markers

Understanding existing levels of genetic variability of camel populations is capital for conservation activities. This study aims to provide information on the genetic diversity of four dromedary populations, including Guerzni, Harcha, Khouari and Marmouri. Blood samples from 227 individuals belonging to the aforementioned populations were obtained and genotyped by 16 microsatellite markers. A total of 215 alleles were observed, with the mean number of alleles per locus being 13.4 ± 6.26. All loci were polymorphic in the studied populations. The average expected heterozygosity varied from a maximum of 0.748 ± 0.122 in Guerzni population to a minimum of 0.702 ± 0.128 in Harcha population; Guerzni population showed the highest value of observed heterozygosity (0.699 ± 0.088), whereas Harcha population the lowest (0.646 ± 0.130). Mean estimates of F-statistics obtained over loci were F_IS = 0.0726, F_IT = 0.0876 and F_ST = 0.0162. The lowest genetic distance was obtained between Guerzni and Khouari (0.023), and the highest genetic distance between Harcha and Marmouri (0.251). The neighbour-joining phylogenetic tree showed two groups of populations indicating a cluster of Guerzni, Khouari and Marmouri, and a clear isolation of Harcha. The genetic distances, the factorial correspondence analysis, the analysis of genetic structure and the phylogenetic tree between populations revealed significant differences between Harcha and other populations, and a high similarity between Guerzni, Khouari and Marmouri. It is concluded from this study that the camel genetic resources studied are well diversified. However, the herd management, especially the random selection of breeding animals, can increase the level of genetic mixing between different populations, mainly among Guerzni, Khouari and Marmouri, that live in the same habitat and grazing area.     

大叶藻居群微卫星遗传多样性研究

采用4对微卫星引物对大叶藻的7个地理居群进行了遗传多样性与遗传结构分析。扩增148株大叶藻得到57个等位基因, 每个位点平均等位基因数为6, 大叶藻居群的平均期望杂合度(He)为0.687, 平均观测杂合度(Ho)为0.417。青岛湾居群的遗传多样性最高(A=7.750, AR=7.043), 俚岛居群最低(A=4.750, AR=4.543)。从F_st值来看, 7个大叶藻居群间属于中度分化。UPGMA系统发育树显示, 中国4个大叶藻居群聚类到一起, 其遗传分化可能是由于历史大海草场的遗留小片段居群产生, 而中国、韩国、日本和爱尔兰居群间的遗传分化则主要是由于地理隔离造成的。自由交配估计结果支持海草的东亚起源说。青岛湾居群遗传多样性较高, 可优先作为大叶藻移植修复的材料和基因库, 并进行重点保护。     

Identification and characterization of microsatellite markers for the population genetic structure in endemic red-tailed barb,Gonoproktopterus curmuca

Gonoproktopterus curmuca is an endangered red tailed barb found in Southern part of Western Ghat, India. As a part of stock-specific, propagation assisted rehabilitation and management program, polymorphic microsatellites markers were used to study the genetic diversity and population structure of this species from the three River systems of Southern Western Ghats, such as Periyar River, the Chalakkudy River, and the Chaliyar River. From selected eight polymorphic microsatellite markers, the number of alleles per locus ranged from 2 to 8, and the average number of alleles among 3 populations ranged from 5.0 to 5.75. The mean observed (H_ob) and expected (H_ex) heterozygosity ranged from 0.5148 to 0.5360 and from 0.5996 to 0.6067, respectively. Significant deviations from Hardy–Weinberg Equilibrium expectation were found at majority of the loci (except Gcur MFW72 and Gcur MFW19) and in all three populations in which heterozygote deficits were apparent. The analysis of molecular variance indicates that the percent of variance among populations and within populations were 6.73 and 93.27, respectively. The pairwise F_ST values between populations indicate that there were significant deviations in genetic differentiations for the red-tailed barb populations from these three Rivers of the Western Ghats, India. The microsatellites methods reported a low degree of gene diversity and lack of genetic heterogeneity in the population of G. curmuca, which strongly emphasize the need of fishery management, conservation and rehabilitation of G. curmuca.     

Population genetic structure of the sea bass (Lateolabrax japonicus) in Korea based on multiplex PCR assays with 12 polymorphic microsatellite markers

Population genetics has been recognized as a key component of policy development for fisheries and conservation management. In this study, natural sea bass (Lateolabrax japonicus) populations in three ocean basins in Korea were assessed using multiplex assays with 12 highly polymorphic microsatellite loci; 203 alleles and similarly high levels of genetic diversity [mean number of alleles (N_A) = 14.43, mean expected heterozygosity (He) = 0.84] were detected. All populations showed significant heterozygote deficiency at four loci, which could be explained by the presence of null alleles. The genetic population subdivision was low and was significantly different according to F-statistics (overall F _ST = 0.003, R _ST = 0.005). However, this substructure was not supported by an analysis of molecular variance test, analyses of isolation by distance or Bayesian analysis. The passive dispersal of eggs/larvae via the main currents appears to facilitate gene flow. The possibility of a recent genetic bottleneck was observed in all three populations of L. japonicus, indicating that overfishing and degradation of the environment in recent years has led to a decline in the sea bass populations in Korea. Our study demonstrates that sea bass in Korea do not appear to be genetically partitioned and should be managed as a single unit; however, the potential for a rapid loss of genetic diversity remains. Information regarding the genetic characteristics of Korean sea bass populations has important implications for fishery management and conservation efforts and will aid in the sustainable exploitation of fishing resources and the preservation of biodiversity.