五个红罗非鱼群体的遗传多样性分析

实验采用微卫星标记技术，选用22对微卫星引物对5个红罗非鱼群体进行遗传多样性分析。经PCR扩增，16个微卫星位点扩增产物在关岛红罗非鱼（GD）、珍珠白红罗非鱼（ZZ）、佛罗里达红罗非鱼（FL）、明月红罗非鱼（MY）、马来西亚红罗非鱼（ML）中均获得了清晰稳定的条带。分析结果显示:16个微卫星标记共检测到146个等位基因。5个群体的平均等位基因数（N_a）在6.5625-8.5625，平均有效等位基因数（N_e）在4.1495-6.1330，平均杂合度（H_e）在0.7491-0.8247，平均多态信息含量（PIC）在0.6939-0.7840，说明它们的遗传多态性丰富。卡方检验表明5个红罗非鱼群体的大部分位点偏离Hardy-Weinberg平衡。在5个群体中，关岛红罗非鱼（GD）与珍珠白红罗非鱼（ZZ）遗传相似系数（0.6171）最小，遗传距离（0.4827）最大，说明两者亲缘关系最远；佛罗里达红罗非鱼（FL）与马来西亚红罗非鱼（ML）遗传相似系数（0.9069）最大，遗传距离（0.0977）最小，可推断两者亲缘关系最近。采用UPGMA进行聚类分析，结果表明:佛罗里达与马来西亚先聚成一支，二者再与珍珠白聚在一起，接着三者与明月聚在一起，最后，四者与关岛聚到一起。聚类结果说明关岛群体与其他4个群体亲缘关系最远；佛罗里达与马来西亚亲缘关系最近，珍珠白群体次之，明月群体再次之。以上结果可推断5个红罗非鱼群体遗传多态性丰富，具有较大的选育潜力。     

虾夷扇贝（Patinopecten yessoensis）5个群体的遗传多样性

虾夷扇贝为20世纪80年代初从日本引入我国并逐渐开展养殖的双壳贝类,目前已在我国北方地区大面积养殖。实验采用微卫星分子遗传标记技术对大连獐子岛底播增殖放流群体（CC）、黄海北部海区采集的野生群体（HQ）、日本青森养殖群体（JX）、俄罗斯远东日本海沿岸养殖群体（RX）及大连大长山岛养殖上壳白化群体（ZB）等5个虾夷扇贝群体的遗传多样性进行研究。其中HQ群体为本课题组2005年在黄海北部采集的野生群体,本研究筛选出一个该群体的特异性遗传标记。用8个微卫星位点进行扩增,共获得45个等位基因,每个位点的等位基因数处于3—9之间,大小为100—340bp,平均有效等位基因数为3．1535,基因型数为3—21个,PIC（PolymorphismInformationContent）值处于0．0322-0．5944之间。5个群体的平均观测杂合度分别为0．3292、0．3048、0．3167、0．2708、0．3042,平均期望杂合度分别为0．4595、0．4002、0．3838、0．3620、0．3885,群体间的多态性差异不显著。根据群体间遗传相似性系数、遗传距离及UPGMA聚类分析发现,CC和HQ群体亲缘关系最近,JX和RX群体的亲缘关系较近,ZB群体与JX和RX群体的亲缘关系较近。通过Hardy—Weinberg平衡及F-检验发现,5个群体都不同程度的偏离平衡,表明各群体基因频率和基因型频率的稳定性较低,且5个群体均处于不同程度的杂合子缺失状态,群体间的遗传分化程度较高,但遗传变异主要来自群体内的个体间。     

东北大口鲇2个群体的微卫星DNA多态分析

利用磁珠富集法克隆制备的24个大口鲇（Silurus meriaionalis Chen）微卫星标记,对黑龙江野生群体与松花江养殖群体2个东北大口鲇（S．soldatovi）的地理种群的等位基因频率（P）、观测杂合度（Ho）、期望杂合度（He）、多态信息含量（PIC）和有效等位基因数（Ne）等进行了遗传检测,以遗传偏离指数（d）检验Hardy—Weinberg平衡,并以Nei氏遗传分化系数（GST）和AMOVA分析（ФST）群体遗传变异的来源。同时,使用PHYLIP3．63软件绘制基于Nei氏遗传距离的个体间UPGMA系统树。结果表明：24个微卫星标记在东北大口鲇的2个群体中共扩增出1357条多态性片段,片段长度为1024385bp,总体平均等位基因8．875个,可以用于东北大口鲇遗传多样性的评估。并发现8个可区分这2个种群的遗传标记;黑龙江群体的P、Ho、He、PIC和Ne依次为0．165、0．435、0．758、0．742和5．019,松花江群体为0．147、0．299、0．847、0．764和5．944,在这些多样性参数上,方差分析也显示2地理种群差异不显著,在大多数位点并无显著差异,仅HLJcf37位点具有显著差异：在多个位点偏离Hardy—Weinberg平衡,2群体呈现不同程度的杂合体过度,纯合体完全缺失现象,其原因有待证实;群体遗传变异分析证实2群体间遗传分化较弱,其98％以上的变异是由群体内个体间的遗传变异引起的,群体间的变异对总变异影响不显著。UPGMA系统树也显示出个体间遗传距离小,亲缘关系很近。结果表明,人工繁殖没有对东北大口鲇的遗传多样性产生影响,该种群遗传分化小,种质资源状况良好。     

吉富罗非鱼雌雄群体遗传差异的SSR分析

为探讨吉富罗非鱼(genetic improvement of farmed tilapia,GIFT)雌、雄群体间的遗传差异,本研究对国家级广西南宁罗非鱼良种场雌、雄吉富罗非鱼进行了遗传差异分析。研究结果表明,选取的11对SSR引物中有10对能获得稳定的目的条带;每个SSR基因座的等位基因数在2～4个之间,雌性罗非鱼的平均等位基因(Na)2.9个,稍高于雄性的2.8个;雌、雄吉富罗非鱼平均观察杂合度(HO)分别为0.4183和0.4154,多态信息含量(PIC)分别为0.4048和0.3932,属中度多态;雌雄个体间的遗传距离和相似性指数分别为0.0908和0.9132。此外,SSR基因座PRL-SO2在雄鱼中偏离Hardy Weinberg平衡(P0.005)。上述结果表明,吉富罗非鱼雌、雄群体的SSR多态性基本相同,推测这两者基因组间的差异较小。     

应用微卫星标DNA记对三个封闭群兔遗传分析

目的检测国内Et本大耳白兔、青紫蓝兔、新西兰白兔的遗传背景及遗传结构,为封闭群兔遗传检测方法建立和标准化提供基础资料。方法应用18个微卫星标记及荧光标记一半自动基因分型技术对三个群体95个个体进行Hardy．Weinberg检测,统计每个位点等位基因频率、杂合度、F值、遗传距离等信息。结果三个种群平均等位基因观测数为3．167、4．556、3．444,平均观测杂合度为0．444、0．5230、0．4976。18个位点平均多态信息含量（PIC）为0．410、0．549、0．470,日本大耳白兔6个位点HWE检验（P〈0．05）,显著偏离Hardy—Weinberg平衡,并在Satl3,INRCCDDV0088位点基因型完全纯和,新西兰白兔和青紫蓝兔分别有2个位点显著偏离Hardy—Weinberg平衡。三个种群遗传距离：青紫蓝兔与新西兰白兔遗传距离最近,为0．124,与日本大耳白兔遗传关系最远,为0．320;新西兰白兔与日本大耳白兔较远,为0：10。结论三个种群有各自不同遗传特征,遗传多样性较高,种群间分化明显。个别位点偏离遗传平衡,推测人工繁育过程中存在一定问题。     

极濒危植物中华水韭休宁居群的遗传结构

采用超薄平板微型聚丙烯酰胺等电聚焦电泳方法对极濒危蕨类植物中华水韭(Isoetes sinensis)现存于安徽休宁的5个亚居群的等位酶多样性和遗传结构进行了研究。结果表明：中华水韭居群每位点平均等位基因数A=1．7,平均多态位点比率P=55．56％,平均预期杂合度He=0．201。居群中半数的多念位点表现为等位基因的“固定杂合”,5个亚居群的遗传多样性无显著差异,但都表现出严重偏离Hardy—Weinberg平衡的杂合子过昔;其遗传变异主要发生于亚居群内(94．27％),亚居群之间的遗传分化较小(Gsl=0．0573),亚居群间遗传一致度较高(I=0．960—0．999)。我们推断这可能是由于居群构建之初的奠基者效应或者原种群曾经历了较为严重的遗传瓶颈所导致;此外亚居群间便利的基因交流也可能起到了很大作用(Nm=4．5062)。取自休宁居群的75株样品由13个专一多位点基因型(18个位点等位酶基因型)组成,亚居群及个体间高度遗传均质。本文还就中华水韭休宁野生居群濒危的遗传因素进行了探讨,并提出了相应的保育策略。     

微卫星标记分析罗非鱼群体的遗传潜力

利用25个微卫星标记,对奥利亚罗非鱼2个群体["夏奥1号"(ZA)、广西群体(GA)]和尼罗罗非鱼4个群体[埃及品系(ZN)、88品系(XN)、广西群体(GN)、美国品系(MN)]进行检测。共检测到7 775个扩增片段,长度在100~400 bp;等位基因数3~8个不等,共计143个等位基因;平均每个基因座扩增得到5.72个等位基因。各群体平均观测杂合度(H o)在0.7253~0.8160之间,平均期望杂合度(He)在0.5146~0.6834之间,平均多态信息含量(PIC)在0.4212~0.6105之间,平均有效等位基因数(A e)在2.20~3.23之间。ZA与GA遗传相似系数最高(0.9130),ZA与ZN遗传相似系数最低(0.4352)。总的说来,4个尼罗罗非鱼群体的遗传潜力较高,2个奥利亚罗非鱼群体的遗传潜力适中。     

利用微卫星标记分析蛋鸡配套系的遗传关系

选用20个微卫星标记分析了2001年和2002年引进的H高产蛋鸡配套系祖代各品系之间的遗传关系,计算杂合度、多态信息含量(PIC)、有效等位基因数、Nei氏遗传距离等统计量,分析了品系内和品系间的遗传变异,并采用非加权类平均法(UPGMA)对各品系进行聚类。结果显示：每个微卫星位点的平均等位基因数为3．250,平均有效等位基因数为2．395,20个微卫星标记共有等位基因65个,各位点的PIC在0．102—0．729之间变动,平均为0．454;各位点杂合度的变动范围是0．108—0．765,各品系杂合度的分布范围是从A2001的0．390到D2001的0．452之间,各品系的杂合度较低,群体内变异较小。配套系各品系是经过高度选育的,所以群体内变异较小,本研究结果与育种实际相符合,也从另一方面说明了品系的遗传特征能够由各微卫星位点的多态性得到真实的反映。A、B两系之间的遗传距离为0．005—0．016,遗传相似系数大于0．984;C、D两系之间的遗传距离为0．094～0．119,遗传相似系数为0．900左右。这些结果表明,A、B应为同一个品系或遗传关系非常近的两个品系,C、D是两个遗传关系较远的品系。     

中国东亚飞蝗两个种群遗传分化的研究

采用水平淀粉凝胶电泳技术,分析了我国河北平山、辽宁葫芦岛两个蝗区东亚飞蝗自然种群的遗传结构。在酶谱分析的19个位点中,多数位点的等位基因数目较少,而位点Aat—l、Pgi和Mdh—2的等位基因数目相对较多。由于杂合子数目较少而使每个基因位点的平均杂合度降低(Ho=0．024和0．028)。对每个位点的各基因型进行x^2检验,绝大多数位点的基因型频率偏离Hardy—Weinberg平衡。较低的Fst值(Fst=0．021)表明两个种群的遗传分化程度不高。两个种群间较高的遗传一致度(I=0．991)和较小的遗传距离(D=0．092)也证实了上述结果。由此推测,该蝗虫较强的迁飞能力有可能增强种群间的基因交流,降低种群间的遗传分化。然而,在一些等位酶指标如位点Ao-2、Fbp—l、Mdh—2的等位基因频率分布、每个位点的平均等位基因数(A=2．5和2．7)和多态位点百分率(P=52．6％和57．9％)等,两个种群之间呈现出明显的差异。这也许与两个种群之间不同的生态环境条件和较远的地理距离有关。     

湖北3种药用淫羊藿植物等位酶多样性与居群遗传结构

采用超薄平板微型聚丙烯酰胺等电聚焦电泳方法对湖北3种淫羊藿属（Epimedium L．）药用植物：柔毛淫羊藿（E．pubescens Maxim．）、箭叶淫羊藿（Esagittatum（Sieb．＆Zucc．）Maxim．）和巫山淫羊藿（Ewushanense T.S．Ying）的11个自然居群进行了等位酶种内遗传多样性和居群遗传结构以及种间遗传关系的研究。6个酶系统在3种淫羊藿植物中检测到13个酶位点共45个等位基因,分析结果表明：1）3种淫羊藿均具有较高的遗传多样性水平（A=2．6-3．2,P=69．2％-84．6％,Ho=0．274-0．377,HE=0．282-0．369）,远高于多年生草本植物和依靠动物传粉植物的平均水平;2）所有居群都显著偏离Hardy—Weinberg平衡,大多数多态位点固定指数（F）明显偏离于0且正负值各半,暗示该属植物可能具有复杂的繁育系统;3）种内和种间的遗传分化度均较低,分别为GST=0．0246-0．0409和0．0495-0．1213,表明种内居群间存在较高的基因流和种间较近的遗传关系;4）聚类分析表明柔毛淫羊藿与箭叶淫羊藿的亲缘关系更近,这与形态学特征、花瓣演化趋势及系统发育分析等方面的研究的结果相吻合。讨论分析认为3种淫羊藿植物的遗传变异特性是由淫羊藿属植物异交与克隆繁殖的混合繁育系统、多年生的生活特性、古老的进化历史和种子的蚁播特性等多因素共同决定的。     

我国新引进吉富品系尼罗罗非鱼群体的遗传多样性分析

选取罗非鱼(Oreochromis spp.)第二代遗传连锁图谱中的26个微卫星位点,对淡水渔业研究中心引进的、由60个家系组成的吉富品系尼罗罗非鱼(O.niloticus)群体进行遗传结构分析。结果显示,26个微卫星位点在吉富罗非鱼群体中共检测到124个等位基因,各位点的等位基因数为3～7个,平均4.8个。片段长度104～322 bp,平均杂合度观测值为0.622 1,平均杂合度期望值为0.642 3,平均多态信息含量(PIC)为0.633 4。所检测的26个位点中,有25个位点属于高度多态位点(PIC0.5),占所检测位点的96.15%;1个位点属于中度多态位点。结果表明,该吉富罗非鱼群体多态信息含量丰富,遗传多样性水平较高。因而该群体仍然具有较大的选育潜力,可以作为选育的基础群体开展进一步的选育工作。     

Isolation and characterization of microsatellite loci for the analysis of genetic diversity in Whitmania pigra

The objective of this study was to isolate microsatellite loci to analyze the genetic diversity of Whitmania pigra. Four new microsatellite markers of W. pigra were developed from an enriched library and ten from a modified SAMPL assay. A total of 127 alleles were detected, with an average of 9.1 alleles per locus. The expected heterozygosity (He) of each microsatellite locus varied from 0.451 to 0.857, with an average of 0.688. The polymorphism information content (PIC) of each microsatellite locus ranged from 0.361 to 0.838, with an average of 0.640. Analysis of molecular variance showed that the main variation component existed within the populations (81.64%) rather than among the populations (18.36%). Phylogenetic tree for 15 populations of Hirudo using the NJ method by MEGA 5.1 software were divided into two major clusters. These microsatellite markers will contribute to research on the individual identification, genetic diversity, population structure, genome mapping and conservation biology of Hirudo.     

Genetic Diversity and Population Structure Analysis Between Indian Red Jungle Fowl and Domestic Chicken Using Microsatellite Markers

The present study was conducted to assess the genetic diversity, population structure, and relatedness in Indian red jungle fowl (RJF, Gallus gallus murgi) from northern India and three domestic chicken populations (gallus gallus domesticus), maintained at the institute farms, namely White Leghorn (WL), Aseel (AS) and Red Cornish (RC) using 25 microsatellite markers. All the markers were polymorphic, the number of alleles at each locus ranged from five (MCW0111) to forty-three (LEI0212) with an average number of 19 alleles per locus. Across all loci, the mean expected heterozygosity and polymorphic information content were 0.883 and 0.872, respectively. Population-specific alleles were found in each population. A UPGMA dendrogram based on shared allele distances clearly revealed two major clusters among the four populations; cluster I had genotypes from RJF and WL whereas cluster II had AS and RC genotypes. Furthermore, the estimation of population structure was performed to understand how genetic variation is partitioned within and among populations. The maximum ?K value was observed for K = 4 with four identified clusters. Furthermore, factorial analysis clearly showed four clustering; each cluster represented the four types of population used in the study. These results clearly, demonstrate the potential of microsatellite markers in elucidating the genetic diversity, relationships, and population structure analysis in RJF and domestic chicken populations.     

Genetic variation in populations of the threatened seagrass Halophila beccarii (Hydrocharitaceae)

Halophila beccarii (Hydrocharitaceae), a small monoecious seagrass, has been listed as a threatened species. In this study, a total of 106 samples were collected from four Chinese populations located at the northern limit of its distribution range. Using six polymorphic microsatellites, we found low genetic variation in this species, in which the mean number of alleles per locus was 2.8, and 16 multi-locus genotypes were revealed. In the four populations, the mean number of alleles per locus ranged from 1.2 to 2, one to eight genotypes were found, and clonal diversity ranged from 0 to 0.23; observed and expected heterozygosity ranged from 0.17 to 0.40 and from 0.09 to 0.24, respectively. Strong genetic differentiation was found among the populations, and the standardized fixation index (F′_ST) was 0.787. Species traits (i.e., clonal growth) and bottleneck effects due to drastic population fluctuation may contribute to the observed low genetic variation.     

Assessing Genetic Diversity in <Emphasis Type="Italic">Olea europaea</Emphasis> L. Using ISSR and SSR Markers

Olea europaea L. is one of the most economically important crops in the Mediterranean area, and known for having large genetic variability. In order to assess the genetic diversity, DNA from 41 olive cultivars, present in the protected denomination of origin (PDO) region of Trás-os-Montes, was screened using inter simple sequence repeat (ISSR) and microsatellite (SSR) markers. Eleven ISSR primers amplified 135 reproducible bands of which 108 were polymorphic. The percentage of polymorphic bands detected by ISSR was 79%. The highest number of polymorphic bands was obtained by the use of primers UBC807 (15) and UBC809 (16). A total of 67 alleles were detected by six SSR primers, with an average of 11 alleles per primer. The number of alleles per locus ranged from five (ssrOeUaDCA05) to 15 (ssrOeUaDCA03). The observed heterozygosity ranged from 0.219 (ssrOeUaDCA05) to 0.900 (ssrOeUaDCA04), while the expected heterozygosity varied between 0.426 (ssrOeUaDCA05) and 0.887 (ssrOeUaDCA03). The polymorphism information content (PIC) values ranged from 0.392 (ssrOeUaDCA05) to 0.863 (ssrOeUaDCA03). The collection of primers selected gave a reasonable number of amplification products for the genetic diversity analysis. Based on the results, the genetic diversity among 41 olive cultivars is discussed. This study reveals the great importance of guaranteeing the differentiation of olive cultivars and their application for certification purposes.     

德宏水牛微卫星标记分析的群体遗传变异

德宏水牛是云南省地方水牛的优良品种之一,为了进一步阐明其群体遗传变异和遗传结构,筛选了分别位于水牛14条染色体上的15对微卫星引物,对德宏水牛81个个体进行了检测分析.共检测到62个等位基因,每个座位等位基因数目从2到6个不等,平均等位基因数为4.13,该水牛群体期望杂合度和多态信息含量分别为0.6520±0.1526和0.5863±0.1789,各座位的遗传分化系数在0～0.0919之间,平均值为0.0202.每个座位的基因流较大,平均12.1502.研究结果表明德宏水牛群体遗传多样性较丰富,亚群间的遗传分化程度低,基因流较大,且很少发生近交.     

三个虹鳟养殖群体遗传结构的微卫星分析

利用20个微卫星标记对3个虹鳟(Oncorhynchus mykiss)养殖群体进行遗传结构分析。结果表明,(1)3个群体检测的平均等位基因数为3·6～4·1,平均观测杂合度为0·5224～0·6328,平均期望杂合度为0·4736～0·5522,平均多态信息含量为0·4354～0·5084,说明这几个群体多态性属于偏高水平,遗传多样性高。通过d值,确定了Hard-Weinberg平衡的偏离情况,发现AY039638、AY039646在3个群体中都表现为不平衡。对3个群体的遗传距离进行了估算,并进行聚类分析,本溪的两个群体先聚为一支,再与渤海群体相聚,显示出明显的地理特征。(2)本溪虹鳟群体在位点AF352770出现部分等位基因消失的现象;AF352754在本溪群体中表现为位点消失,可作为区分本溪群体和渤海群体的分子标记。(3)综合评价3个群体,渤海站虹鳟群体的遗传多样性最高,与前人研究结果一致。     

Development of microsatellite markers to genetically differentiate populations of Octopus minor from Korea and China

Of the more than 300 octopus species, Octopus minor is one of the most popular and economically important species in Eastern Asia, including Korea, along with O. vulgaris, O. ocellatus, and O. aegina. We developed 19 microsatellite markers from Octopus minor and eight polymorphic markers were developed to analyze the genetic diversity and relationships among four octopus populations from Korea and three from China. The number of alleles per locus varied from 10 to 49, and allelic richness per locus ranged from 2 to 16.4 across all populations. The average allele number among the populations was 11.1, with a minimum of 8.3 and a maximum of 13.6. The mean allelic richness was 8.7 in all populations. The Hardy-Weinberg equilibrium (HWE) test revealed significant deviation in 19 of the 56 single-locus sites, and null alleles were presumed in five of eight loci. The pairwise F ( ST ) values between populations from Korea and China differed significantly in all pairwise comparisons. The genetic distances between the China and Korea samples ranged from 0.161 to 0.454. The genetic distances among the populations from Korea ranged from 0.033 to 0.090, with an average of 0.062; those among populations from China ranged from 0.191 to 0.316, with an average of 0.254. The populations from Korea and China formed clearly separated into clusters via an unweighted pair group method with arithmetic mean dendrogram. Furthermore, a population from muddy flats on the western coast of the Korean Peninsula and one from a rocky area on Jeju Island formed clearly separated subclusters. An assignment test based on the allele distribution discriminated between the Korean and Chinese origins with 96.9 % accuracy.     

利用微卫星分子标记分析四个中华绒螯蟹群体的遗传多样性

利用本实验室克隆的16个和国际上发表的8个微卫星标记,对4个中华绒螯蟹群体(江苏、安徽、辽宁、天津)的遗传多样性进行检测。所检测到的扩增片段长度为80—445bp,在群体间扩增出2—10个等位基因,共计155个等位基因,平均等位基因6.458个。4个中华绒螯蟹群体的平均有效等位基因数(Ne)为4.3491—4.7234,平均观察杂合度(Ho)为0.5690—0.6722,平均期望杂合度(He)为0.7238—0.7546,并通过基因型的P值,确定了7个座位处于Hardy-Weinberg平衡;同时对4个群体的遗传距离进行了估算,聚类分析结果表明,安徽、江苏、天津聚为一支,属于长江河蟹类型,辽河种群单独聚为一支。