黄鳝减数分裂雌核发育及子代遗传纯合度分析

为了培育性状优良、遗传稳定的黄鳝(Monopterus albus)群体,研究探索了人工诱导黄鳝减数分裂雌核发育方法。针对养殖性状优良的深黄大斑鳝进行种质纯合,创制出3个深黄大斑鳝雌核发育群体。流式细胞术和染色体计数分析表明雌核发育黄鳝的细胞DNA含量、染色体数目均与野生型相同。性腺组织学切片观察表明雌核发育黄鳝卵巢发育正常。微卫星多样性分析表明雌核发育黄鳝群体的有效等位基因(N_e)、平均香农指数(I)、平均多态信息指数(PIC)、平均观测杂合度(H_o)及平均期望杂合度(H_e)等参数均极显著低于野生群体,雌核发育群体的遗传纯合度显著提高。雌核发育群体之间的遗传距离增大,遗传相似系数变小。深黄大斑鳝雌核发育群体为培育性状优良的黄鳝养殖品种提供了育种材料。     

草鱼连续两代的雌核发育群体及湘江流域群体基因组DNA的微卫星比较分析

对湘江流域草鱼群体,一代及连续两代极体型人工诱导雌核发育草鱼群体基因组DNA的微卫星引物统计分析结果表明:湘江流域草鱼群体存在一定程度的遗传多态性;大多数被检测的微卫星位点上存在两个以上的等位基因,但遗传多样性程度较低。一代人工诱导雌核发育草鱼群体中个体的基因位点已基本纯合,但就整个群体而言,个体之间的基因型还不完全一致,表现出一定的多态性。连续两代人工诱导雌核发育草鱼群体中不仅所检测个体的基因位点已完全纯合,并且各个体的基因型也完全相同。这些观察结果说明所检测的两代人工诱导雌核发育草鱼群体是纯合的,经连续两代人工诱导雌核发育可能建立起草鱼纯系。该实验结果还发现不仅草鱼的微卫星位点上存在等位基因的多态性,而且微卫星位点本身也存在多态性;在人工诱导草鱼雌核发育的过程中不仅存在微卫星等位基因快速丢失的现象,而且也存在微卫星位点丢失的现象。因此,加强对自然水体中草鱼种质资源多样性的保护和利用各种现代生物学技术纯化、筛选和组合优良性状基因,是草鱼遗传育种中同样重要和不可或缺的两个方面。     

两个不同的人工雌核发育草鱼群体基因组DNA的RAPD分析

采用随机扩增多态性DNA(RAPD)技术对连续两代人工诱导雌核发育草鱼群体和一代人工诱导雌核发育草鱼群体的群体内的遗传相似度、遗传距离以及群体多样性进行了分析 ,并用一个随机取样的普通草鱼群体作比较。用 2 6个多态性引物在 3个群体中共检测到了 2 91个扩增位点 ;在 3个群体中检测到的位点数分别为 2 6 5、2 72和2 82。其中多态性位点数分别为 15、19和 81。遗传学统计分析结果表明 :3个群体的多态位点比例分别为 5 6 6 %、6 99%、2 8 72 % ;香农表型多样性指数分别为 0 170 2、0 316 9和 0 84 5 0 ;按照Nei指数统计的三个群体的遗传相似度分别为 0 985 1、0 982 0、0 9114。这些结果表明 :两个雌核发育草鱼群体的遗传多样性远低于普通草鱼群体 ,而其群体的基因组DNA同质性远高于普通草鱼。而在两个雌核发育草鱼群体中 ,两代雌核发育群体的遗传多样性要低于一代雌核发育群体的遗传多样性 ;而群体的遗传纯合度则前者高于后者。     

翘嘴红鲌(Erythroculteri lishaeformis)精子诱导的雌核发育团头鲂(Megalobrama amblycephala)细胞及其分子遗传学分析

该研究运用紫外灭活的翘嘴红鲌(Erythroculter ilishaeformis)精子刺激团头鲂(Megalobrama amblycephala)卵子进行雌核发育,并于0~4℃冷水刺激条件下获得雌核发育团头鲂群体。在此基础上,于细胞遗传学(DNA含量测定、染色体数目检测及性腺发育观察)及分子遗传学水平(微卫星分析)研究了雌核发育团头鲂的细胞生物学以及遗传学性状。结果表明,雌核发育团头鲂DNA含量与普通团头鲂一致(2n=48),其外形特征与对照组极为相似,且均为雌性,为团头鲂性别决定方式(XY型)提供了细胞遗传学依据。同时,微卫星结果表明,在普通团头鲂、雌核发育团头鲂及翘嘴红鲌3个群体中共扩增出63个等位基因,雌核发育团头鲂群体平均观测杂合度和平均期望杂合度均显著低于亲本,即经过一代雌核发育,雌核发育团头鲂基因纯合度显著高于普通团头鲂和翘嘴红鲌,已达到快速建立纯系的目的,且雌核发育团头鲂群体与普通团头鲂群体遗传距离接近,即雌核发育为母系遗传。该研究为团头鲂遗传选育及品系改良提供了遗传材料。     

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。     

红白锦鲤人工雌核发育纯系的微卫星标记分析

利用连续两代人工雌核发育操作,构建了极具观赏价值的红白锦鲤人工雌核发育系。应用6对微卫星引物进行遗传标记研究,检测分析了该雌核发育系内个体间的遗传同质性及其基因座位的纯合度。实验结果显示,11尾经人工挑选具有相同表形、极具观赏价值的F1代性成熟个体的微卫星扩增图谱呈现出高度的一致性。同时,随机挑选的人工雌核发育F2代个体在所检测的基因座位均呈现纯合,扩增图谱呈现了高度的一致,从分子水平上证实了本实验所获得的红白人工雌核发育F2代是一个纯系。     

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。     

墨西哥湾扇贝与“中科红”海湾扇贝群体遗传多样性SSR分析

利用微卫星(SSR)分子标记技术,对墨西哥湾扇贝和"中科红"海湾扇贝2个群体共80个个体的遗传结构和遗传多样性进行分析。结果显示:6个微卫星位点共扩增出31个等位基因,各位点的等位基因数为4~8个,平均等位基因数为5.2个;墨西哥湾扇贝与"中科红"海湾扇贝群体平均有效等位基因数(Ne)分别为2.890、2.753;平均观测杂合度(Ho)分别为0.415、0.353;平均期望杂合度(He)分别为0.604、0.479;多态信息含量(PIC)分别为0.554、0.444;两群体具有相同的等位基因数(Na)(4.333)。Hardy-Weinberg平衡检验发现,2个群体各有3个位点(50%)偏离平衡(p0.05),且均表现为杂合子缺失。两群体的遗传分化指数(Fst)为0.109 7,遗传距离(Dxy)为0.316 4,遗传相似性系数为0.728 8。研究结果说明:两个群体均保持较高的遗传多样性,但经过多年的定向选育,"中科红"海湾扇贝群体的遗传多样性低于墨西哥湾扇贝群体且两个群体具有中等程度的遗传分化,存在一定的遗传差异。     

对中国农业大学鸡资源群进行基因组扫描的初步分析

利用PCR结合荧光半自动检测技术对中国农业大学鸡资源家系的 1个包括 4 4 0个个体的 3代群体进行了大规模基因组扫描 ,分析了 5 5个微卫星标记位点。经家系分析表明 ,这些位点都符合孟德尔共显性遗传规律 ,其杂合度在 0～ 0 86之间 ,其中 72 %的位点的杂合度 >0 6 0 ;多态信息含量在 0～ 0 88之间 ,73%的位点的PIC >0 5 0。A系和C系群体在 5 5个位点中的绝大多数位点上存在非共有等位基因 ,两个品系在大多数位点的等位基因和等位基因频率差异显著 (P <0 0 5 ) ,许多位点差异达到极显著水平 (P <0 0 1) ;C系显示出更高的群体整齐度。利用 5 5个微卫星位点的等位基因频率 ,计算出A系和C系的奈氏群体间相似系数I和标准遗传距离D分别为0 10 0 2和 0 892 8。     

金黄地鼠封闭群SPF化后的微卫星遗传学检测

目的检测和评估金黄地鼠封闭群SPF化后的遗传学变化,为SPF金黄地鼠遗传质量的控制提供技术资料。方法应用小鼠和大鼠的微卫星标记筛选适于金黄地鼠遗传检测的微卫星标记,并结合微卫星荧光标记-半自动基因分型技术,对成都生物制品研究所的SPF级金黄地鼠及其来源的普通级金黄地鼠进行遗传检测,计算其群体遗传学参数。结果对18个小鼠和6个大鼠微卫星标记进行了筛选,分别有2个小鼠和2个大鼠微卫星标记在金黄地鼠种群中具PCR扩增多态性。4个检测的微卫星位点在普通级金黄地鼠和SPF金黄地鼠种群分别发现25和20个等位基因,两群体的期望杂合度分别为0.4979和0.5048,其群体遗传多样性无显著差异;群体间的不同微卫星位点FST范围从0.0095到0.0367,平均为0.0315,表明两群体间的遗传分化很弱,其遗传多样性主要存在于群体内;Nei(1972)遗传距离和Nei(1978)无偏遗传距离分别为0.0678和0.0570,表明了2群体之间很高的遗传相似度和非常近的亲缘关系;Hardy-Weinberg平衡检验表明普通级和SPF金黄地鼠分别有2个和3个位点偏离遗传平衡,且偏离位点均表现为杂合子缺陷。结论该SPF金黄地鼠基本保持了其来源普通级黄地鼠的遗传多样性,两群体间遗传分化程度和遗传差异很小,但应进一步加强其封闭群的繁育控制,保持其遗传稳定性。     

Microsatellite–Centromere Mapping in Large Yellow Croaker (<Emphasis Type="Italic">Pseudosciaena crocea</Emphasis>) Using Gynogenetic Diploid Families

The large yellow croaker (Pseudosciaena crocea) is an economically important marine fish in China. Inheritance of 22 heterozygous microsatellite loci was examined in normal crossed diploid families and meio-gynogenetic families in P. crocea. Two gynogenetic families were produced via inhibition of the second polar body in eggs fertilized with UV-irradiated sperm. The ratio of gynogenesis was proven to be 100% and 96.9% in the two families, respectively. Of the 22 examined loci, 4 showed a segregation distortion in both control and gynogenetic families. Microsatellite–centromere (M–C) map distances were examined using 18 loci with normal Mendelian segregation. Estimated recombination rates ranged between 0 and 1.0 under the assumption of complete interference. High recombinant frequencies between heterozygous markers and the centromere were found in large yellow croaker, as in other teleosts. The average recombination frequency was 0.586. Ten loci showed high M–C recombination with frequency greater than 0.67. M–C distances provide useful information for gene mapping in large yellow croaker.     

雌核发育团头鲂的形态和遗传特征分析

利用团头鲂(Megalobrama amblycephala)正常二倍体群体作为对照组, 对团头鲂减数分裂雌核发育二倍体群体的形态特征、染色体组型、性腺发育及遗传特征进行了分析. 结果表明: 雌核发育群体与正常群体在外部可数、可量性状上没有显著性差异(P0.05); 但雌核发育群体出现了尾鳍条数为12的畸形个体, 与正常个体之间有较大的差异; 两个群体的染色体条数都是48, 核型均为18 m+26 sm+4 st; 观察了20尾雌核发育个体的性腺, 均为雌性个体且卵巢发育良好; 采用10个微卫星标记对2个群体的遗传多样性分析, 结果表明正常群体和雌核发育群体平均等位基因数分别为3.8个和1.7个, 雌核发育群体的多态性显著低于正常群体, 表明减数分裂雌核发育二倍体具有高度的遗传相似性, 可作为一个很好的育种材料.       

养殖大黄鱼一个繁育群体亲本亲缘关系剖析

该文利用23个微卫星标记对103尾大黄鱼繁育亲本进行遗传多样性检测和亲缘关系重建分析,并构建繁育体系指导大黄鱼配组。遗传多样性检测显示,103尾亲鱼在23个座位共获得等位基因数134个,平均5.82个,总平均观察杂合度0.599 3,表明该繁育群体尚保持一定水平的遗传多样性。采用似然率和组合优化法统计模型重建的同胞关系不尽相同,但结果均证实了这些繁育亲本亲缘关系十分相近。配组比对分析结果显示,两种方法的配组方案一致性高达85%,最终选择组合优化法的分组方案指导大黄鱼配组繁育。     

大黄鱼快长相关微卫星标记的筛选与验证

为了筛选与大黄鱼生长性状紧密相关的分子标记,并对这些标记的有效性进行鉴定,研究以大黄鱼家系和群体为材料,经标记筛选、两次验证共三个步骤,找到2个与大黄鱼生长性状紧密相关的微卫星标记:LYC0088和LYC0143,其中LYC0088与体高、体质量均极显著相关（P0.01）,与体长显著相关（P0.05）,LYC0143与体长和体质量显著相关（P0.05）,与体高的相关性极显著（P0.01）。LYC0088与LYC0143位点的优势等位基因分别为E、A,优势等位基因型均为AB,优势等位基因型组合为AB/AB,两个位点呈加性效应。该结果为开展大黄鱼分子标记辅助育种提供了有价值的遗传标记。       

Lack of genetic structure in endangered large yellow croaker Larimichthys crocea from China inferred from mitochondrial control region sequence data

The large yellow croaker Larimichthys crocea is an important commercial marine fish species in China. However, information about the population structure of this species is limited. In the present study, mitochondrial DNA (mtDNA) control region was sequenced from four populations of the yellow croaker in the southern Yellow Sea, East China Sea and South China Sea to investigate the genetic structure of this species. A total of 54 haplotypes were detected among 62 individuals of large yellow croaker. High levels of population genetic diversity were observed. Among the four populations, the haplotype diversity was between 0.9895 ± 0.0193 (Xiamen) and 1.0000 ± 0.0524 (Lvsi, Zhoushan). The nucleotide diversity ranged from 0.0208 ± 0.0108 (Xuwen) to 0.0246 ± 0.0138 (Lvsi). The results of AMOVA detected no significant differences among populations. The conventional F_ST statistics were negative and insignificant values. These indicated lack of population genetic structure throughout the Yellow Sea, East China Sea and South China Sea, and random mixing of individuals among the samples. Biological characters of large yellow croaker and lack of physical barrier in the studied area could be the reasons for lack of genetic structure in this species.     

人工雌核发育鲢的遗传多样性及异源遗传物质整入的RAPD分析

运用RAPD技术对连续二代人工雌核发育鲢的遗传多样性及异源遗传物质的整入进行了分析 ,结果表明 :一代雌核发育鲢 ,个体间遗传相似度为 0 94 5— 0 995 6 ,多样性指数为 0 175 ;二代雌核发育鲢 ,个体间遗传相似度为0 96 15— 1 0 0 ,平均为 0 985 2 ,多样性指数为 0 0 6 2。研究揭示经过连续二代人工雌核发育后 ,其遗传多样性明显减少 ,种质进一步纯化。通过对雌核发育鲢二代、亲本鲢和雄鲤的RAPD扩增比较 ,发现雌核发育鲢含有少数与父本相同的特异DNA扩增带 ,而亲本鲢没有 ,在基因水平上表明雌核发育鲢整入了雄鲤的遗传物质     

The similarity index and DNA fingerprinting

DNA-fingerprint similarity is being used increasingly to make inferences about levels of genetic variation within and between natural populations. It is shown that the similarity index--the average fraction of shared restriction fragments--provides upwardly biased estimates of population homozygosity but nearly unbiased estimates of the average identity-in-state for random pairs of individuals. A method is suggested for partitioning the DNA-fingerprint dissimilarity into within- and between-population components. Some simple expressions are given for the sampling variances of these estimators.     

Microsatellite analysis reveals genetically distinct populations of red pine (Pinus resinosa, Pinaceae)

Red pine (Pinus resinosa Ait.) is an ecologically and economically important forest tree species of northeastern North America and is considered one of the most genetically depauperate conifer species in the region. We have isolated and characterized 13 nuclear microsatellite loci by screening a partial genomic library with di-, tri-, and tetranucleotide repeat oligonucleotide probes. In an analysis of over 500 individuals representing 17 red pine populations from Manitoba through Newfoundland, five polymorphic microsatellite loci with an average of nine alleles per locus were identified. The mean expected and observed heterozygosity values were 0.508 and 0.185, respectively. Significant departures from Hardy-Weinberg equilibrium with excess homozygosity indicating high levels of inbreeding were evident in all populations studied. The population differentiation was high with 28-35% of genetic variation partitioned among populations. The genetic distance analysis showed that three northeastern (two Newfoundland and one New Brunswick) populations are genetically distinct from the remaining populations. The coalescence-based analysis suggests that "northeastern" and "main" populations likely became isolated during the most recent Pleistocene glacial period, and severe population bottlenecks may have led to the evolution of a highly selfing mating system in red pine.     

Population structure of large yellow croaker (Larimichthys crocea) revealed by single nucleotide polymorphisms

The large yellow croaker, Larimichthys crocea, is one of the most commercially important marine species native to China Seas. In this study, Sequenom MassARRAY was used to analyze 120 individuals from three geographic populations in the East and South China Seas and a cultured stock. Forty-four of 60 single nucleotide polymorphism (SNP) loci were ascertained. Eleven loci, scf117, scf708, scf247, scf595, scf100, scf166, scf305, scf664, scf117, scf187 and scf511 each showed population-specific genotypes. A total of 127 alleles were detected. Averagely, the effective number of alleles was between 1.328 and 1.341, expected heterozygosity and observed heterozygosity ranged from 0.273 to 0.320, and 0.190 to 0.235, respectively, Pic value ranged from 0.215 to 0.247, and F_IS ranged from 0.174 to 0.279. The neighbor-joining tree based on calculated genetic distances showed a concordant topology with the traditional morphological characterization. When the population size of wild large yellow croaker drastically has shrunk over the past 20 years, the genetic diversity in both wild and farmed stocks has declined as well.