罗非鱼4个选育群体遗传结构SSR分析

采用SSR分子标记分析了吉富品系罗非鱼的两个家系(GF1和GF2)以及奥利亚罗非鱼(Fo)和奥本尼罗罗非鱼(Fn)群体的遗传结构。结果显示,扩增后等位基因数为3～8个,随引物不同而异,14对引物共扩增60个等位基因,扩增片断大小在102～267 bp之间。微卫星分析表明,奥本尼罗罗非鱼(Fn)的平均观测杂合度(0.764 3)和平均期望杂合度(0.519 6)均最高,吉富尼罗罗非鱼(GF1)平均多态信息含量(0.419 5)最高;吉富尼罗罗非鱼(GF2)平均观测杂合度(0.614 2),奥利亚罗非鱼(Fo)的平均期望观测度(0.446 6)、平均多态信息含量(0.359 2)最低;因此,吉富鱼(GF1)的遗传多样性最高,奥利亚的遗传多样性最低。Hardy-Weinberg平衡遗传偏离指数(D)奥利亚(Fo)和尼罗(Fn)(0.463 4和0.478 9)明显高于吉富的两家系(0.234 1和0.250 0)。Fo与GF2间遗传距离(0.477 0)最大;而GF1和GF2间的遗传距离(0.302 7)最小,遗传相似系数(0.607 3)最大,可推断新一代吉富罗非鱼与本地选育群体有相对较远的亲缘关系,更具杂种优势。     

流沙湾卵鳎微卫星标记的遗传多样性分析

为分析湛江流沙湾海域优势渔种卵鳎的遗传多样性,应用微卫星标记技术,选用15对微卫星引物,以等位基因数、基因杂合度、多态信息含量、固定指数等遗传参数为指标,评估卵鳎群体内的遗传多态性。结果表明:共检测到90个等位基因,等位基因数从1~12不等,平均为6.0;有效等位基因数从1.0~8.4,平均为4.0,多态性位点比例为53%,显示其具有中等杂合子水平,其中8个多态位点的期望杂合度(He)为0.670~0.881,平均为0.800,观测杂合度(Ho)为0.353~1.000,平均为0.773,多态信息含量(PIC)值为0.616~0.870,平均为0.773,群体内固定指数F为-0.199~0.564,平均为0.046;流沙湾卵鳎群体具有高度遗传多样性。     

基于近缘EST-SSR引物的企鹅珍珠贝养殖和野生群体的遗传标记

利用近源物种马氏珠母贝的EST-SSR引物对企鹅珍珠贝DNA进行扩增,获得7对可扩增出目标条带的近缘分子标记,分析野生群体(YS)和养殖群体(YZ)企鹅珍珠贝的遗传多样性。结果显示,7个位点共获得26个等位基因;野生和养殖群体的等位基因数分别为3~5和3~4,平均期望杂合度(He)分别为0.623 5、0.496 8;平均观测杂合度(Ho)分别为0.431 8、0.302 8;HNUPM047位点的多态信息含量较低,为0.446 7,其他位点均在0.5以上,说明该遗传标记可提供比较丰富的遗传信息。表明养殖群体遗传多样性水平低于野生群体。     

大口黑鲈选育群体遗传结构的微卫星分析

采用微卫星标记技术分析不同世代大口黑鲈选育群体的遗传结构。利用11对微卫星引物对大口黑鲈第2～4代选育群体及南水群体(对照)共120个个体进行PCR扩增,共得到28个等位基因。每个位点获得2～3个等位基因,平均等位基因为2.54。第2代(F_2)、第3代(F_3)、第4代(F_4)及南水群体(CG)的平均多态信息含量(PIC)值分别为:0.423、0.419、0.386、0.366。与F_2相比,F_4的遗传多样性减少8.76%,但与CG相比,F_4仍具有较高的遗传多样性,表明大口黑鲈选育群体的遗传基础逐步趋向纯化,且仍有较大选育潜力。此外,尽管F_2与F_4的遗传距离(0.022)比F_2与F_3的(0.011)大,但世代间的F_(st)值(0.006～0.009)均小于0.05,且世代群体总遗传分化指数为0.013,表明选育群体已出现遗传分化,但分化程度较低。     

胡子鲇3个养殖群体遗传结构的AFLP分析

应用AFLP方法,从64对引物中筛选出5对用于分析胡子鲇(Claris fuscus)厦门(XM)、江门(JM)、玉林(YL)等3个养殖群体共90个个体群体内、群体间遗传多样性。扩增共得到289条清晰谱带,引物多态性介于47.8%～63.8%,平均多态性比率为56.4%。胡子鲇群体总遗传杂合度(Ht)、遗传分化系数(Hs)、遗传分化度(Gst)和基因流(Nm)分别为0.161 1、0.153 6、0.051 1和8.501 8,表明胡子鲇多态性处于中等偏下水平,群体内部遗传分化系数较高,三个群体间无较大分化,群体间基因流动较好。其中,JM群体多态性比率、有效等位基因数Ne、Nei’s指数和Shannon指数都较XM群体和YL群体高,种内分化程度较好。对群体进行两两比较发现,XM群体和YL群体间分化程度最大,XM群体和JM群体次之,JM群体和YL群体最小。基于杂交优势,推断XM和YL群体繁殖所产生的后代将具有比亲代更为优良的性状。     

军曹鱼全人工繁殖群体遗传特征的SSR分析

利用8个微卫星DNA位点分析南海海域5个军曹鱼全人工繁育群体(HN1、HN2、ZJ、FJ和LS)子代的遗传多样性特征和群体间遗传分化。结果显示,军曹鱼养殖群体与天然群体的遗传结构特征基本一致:1)平均有效等位基因数为3.910±0.440,观测杂合度为0.595±0.049,分子方差分析(AMOVA)表明,个体内分化占主导(46%),军曹鱼养殖群体整体遗传多样性较高;2)群体间基因流明显(N_m=2.5959,F_(st)=0.0878),整体分化程度较低。各养殖群体表现出不同于天然大群体的特征:1)绝大部分位点均明显偏离哈温平衡,杂合子缺失或过剩现象普遍存在;2)聚类和群体分配分析等表明HN2与另四个养殖群体(HN1、ZJ、FJ和LS)分化明显。     

马氏珠母贝4种壳色家系遗传差异的SSR分析

采用6对微卫星分子标记,分析了马氏珠母贝(Pinctada martensii)白、黑、黄、红4种壳色家系的遗传结构。结果表明:4种壳色家系在6个位点的平均等位基因数为4.17,有效等位基因数为1.297~1.819;平均观测杂合度为0.083~0.212,平均期望杂合度为0.207~0.341;Shannon多样性指数为0.366~0.616;平均多肽信息(PIC)含量为0.257~0.442;4个壳色家系哈-温平衡指数均发生了极显著的偏离,4个家系间的群体再分效应的固定指数FST值为0.113~0.793;与马氏珠母贝普通养殖群体及壳色选育系F5相比,4种壳色家系之间遗传距离变大,遗传多样性降低。     

黃鳍棘鲷放流苗种的遗传质量评估

【目的】评估黃鳍棘鲷(Acanthopagruslatus)增殖放流苗种的遗传质量。【方法】对黃鳍棘鲷放流苗种和自然群体的遗传多样性水平、遗传分化和近交程度等进行对比分析。【结果】放流苗种群体的等位基因数目NA(均值为9.917)大于自然群体NA(均值为8.083);而放流苗种群体的观测杂合度HO(均值为0.646)、期望杂合度HE(均值为0.694)和多态信息含量PIC(均值为0.654)均小于自然群体的HO(均值为0.651)、HE(均值为0.707)和PIC(均值为0.659)。此外,放流苗种群体和自然群体之间的存在一定的遗传分歧,遗传分化指数FST为0.011,但分歧程度不高,且近交程度系数均值F和FIS均大于自然群体的对应系数。【结论】黃鳍棘鲷放流苗种存在一定遗传质量缺陷,不能满足增殖放流要求,可能会自然群体产生负面影响。     

两个凡纳滨对虾亲本群体的RAPD研究

采用Operon公司的 16个引物对两批凡纳滨对虾亲虾的基因组DNA多态性进行了检测 ,结果表明 ,第一个亲本群体对 16个引物共检测到 78个位点 ,其中多态位点数为 4 8个 ,占 6 1.5 4% ;第二个亲本群体共获得 97个位点 ,其中多态位点数为 4 9个 ,占 5 0 .5 2 %。单个引物获得的标记数为 1～ 8个。第一个亲本群体个体间的平均遗传距离为 0 .196 0± 0 .0 392 ,第二个亲本群体个体间的平均遗传距离为 0 .0 92 2± 0 .0 189。两个亲本群体间的多态位点比例为 6 6 .98% ,遗传距离为0 345 5± 0 .0 795。在OPF 0 9、OPV 19、OPZ 113个引物的扩增结果中 ,发现两个亲本群体有明显差异。     

5种虾虎鱼类线粒体COI基因序列变异及系统进化

获得雷州半岛红树林海区5种虾虎鱼稚幼鱼的45条线粒体COI基因序列,其T、C、A、G碱基平均含量分别为29.9%、28.4%、23.9%、17.8%。45条序列共定义23个单倍型,其中检测到变异位点184个,双带缟虾虎鱼(Tridentiger bifasciatus)单倍型比例最高(80.0%),湖栖鳍虾虎鱼(Gobiopterus lacustris)单倍型比例最低(18.2%)。Kimura 2-parameter遗传距离显示,种间遗传距离为0.137 6~0.263 5,种内遗传距离为0.000 0~0.020 8。NJ进化树及ML进化树均表明,聚为5大支系的虾虎鱼类中,阿部氏鲻虾虎鱼(Mugilogobius abei)与诸氏鲻虾虎鱼(M.chulae)亲缘关系最近,湖栖鳍虾虎鱼与其他4种虾虎鱼的亲缘关系稍远。基于群体内等位基因频率分布的中性检验结果表明,双带缟虾虎鱼(Tridentiger bifasciatus)、小口拟虾虎鱼(Pseudogobius masago)、阿部氏鲻虾虎鱼、诸氏鲻虾虎鱼4个群体存在大量低频等位基因位点,而湖栖鳍虾虎鱼群体以中等频率等位基因为主。     

Analysis of genetic diversity and differentiation of seven stocks of Litopenaeus vannamei using microsatellite markers

Seven microsatellite markers were used to evaluate the genetic diversity and differentiation of seven stocks of Litopenaeus vannamei, which were introduced from Central and South America to China. All seven microsatellite loci were polymorphic, with polymorphism information content(PIC) values ranging from 0.593 to 0.952. Totally 92 alleles were identified, and the number of alleles(Na) and effective alleles(Ne) varied between 4 and 21 and 2.7 and 14.6, respectively. Observed heterozygosity(Ho) values were lower than the expected heterozygosity(He) values(0.526–0.754), which indicated that the seven stocks possessed a rich genetic diversity. Thirty-seven tests were detected for reasonable significant deviation from Hardy-Weinberg equilibrium. Fis values were positive at five loci, suggesting that there was a relatively high degree of inbreeding within stocks. Pairwise Fst values ranged from 0.0225 to 0.151, and most of the stock pairs were moderately differentiated. Genetic distance and cluster analysis using UPGMA revealed a close genetic relationship of L. vannamei between Pop2 and Pop3. AMOVA indicated that the genetic variation among stocks(11.3%) was much lower than that within stocks(88.7%). Although the seven stocks had a certain degree of genetic differentiation and a rich genetic diversity, there is an increasing risk of decreased performance due to inbreeding in subsequent generations.     

Genetic variation in two sea cucumber （Apostichopus japonicus） stocks revealed by ISSR markers

Sea cucumber Apostichopus japonicus samples were collected in Changdao, Penglai (PL), 27 individuals, and Lingshandao, Qingdao (QD), 30 individuals, in the Shandong Peninsula, China. Ten SSR primers were used to assess the genetic variation and relationship between and within the two stocks. Respectively, for each stock, the percentage of polymorphic bands was 85.2% and 86.9%; the gene diver- sity was 0.360 5 and 0.342 8; and the Shannon’s information index was 0.515 0 and 0.499 0. At species level, the percentage of polymorphic bands was 92.2%, the total gene diversity was 0.378 9 and the Shannon’s information index was 0.550 4. The coefficient of overall genetic differentiation and the ge- netic distances between the stocks were also calculated to be 0.073 0 and 0.079 6 using the POPGENE program. Results show that the genetic diversity of the two stocks is still large but the genetic distance between the two stocks is close. A dendrogram was constructed for the 57 individuals from the two stocks, showing that the genetic structure was unitary for PL stock but complex for QD stock.     

Genetic diversity in two Japanese flounder populations from China seas inferred using microsatellite markers and COI sequences

Japanese flounder is one of the most important commercial species in China; however, information on the genetic background of natural populations in China seas is scarce. The lack of genetic data has hampered fishery management and aquaculture development programs for this species. In the present study, we have analyzed the genetic diversity in natural populations of Japanese flounder sampled from the Yellow Sea (Qingdao population, QD) and East China Sea (Zhoushan population, ZS) using 10 polymorphic microsatellite loci and cytochrome c oxidase subunit I (COI) sequencing data. A total of 68 different alleles were observed over 10 microsatellite loci. The total number of alleles per locus ranged from 2 to 9, and the number of genotypes per locus ranged from 3 to 45. The observed heterozygosity and expected heterozygosity in QD were 0.733 and 0.779, respectively, and in ZS the heterozygosity values were 0.708 and 0.783, respectively. Significant departures from Hardy-Weinberg equilibrium were observed in 7 of the 10 microsatellite loci in each of the two populations. The COI sequencing analysis revealed 25 polymorphic sites and 15 haplotypes in the two populations. The haplotype diversity and nucleotide diversity in the QD population were 0.746±0.072 8 and 0.003 34±0.001 03 respectively, and in ZS population the genetic diversity values were 0.712±0.047 0 and 0.003 18±0.000 49, respectively. The microsatellite data (F_st =0.048 7, P<0.001) and mitochondrial DNA data (F_st =0.128, P<0.001) both revealed significant genetic differentiation between the two populations. The information on the genetic variation and differentiation in Japanese flounder obtained in this study could be used to set up suitable guidelines for the management and conservation of this species, as well as for managing artificial selection programs. In future studies, more geographically diverse stocks should be used to obtain a deeper understanding of the population structure of Japanese flounder in the China seas and adjacent regions.     

Genetic diversity in old populations of sessile oak from Calabria assessed by nuclear and chloroplast SSR

In the framework of forest resources conservation, this study aims to understand the dynamic and the genetic structure of sessile oak forests in Calabria, Italy. Two old populations of sessile oak(Quercus petraea(Mattuschka) Liebl.) from two areas of Sila and Aspromonte massifs in Calabria were analyzed for genetic diversity and population structure based on 6 nuclear simple sequence repeat(nSSR) and 4 chloroplastic SSR(cpSSR) loci. The populations displayed high amount of genetic diversity, which was toughly structured according to their geographical origins. Number of alleles at SSR loci ranged from 11 to 20 with an average of 13.5 per locus. Gene diversity(expected heterozygosity, He) estimates ranged from 0.575 to 0.834 with a mean of 0.749. The observed heterozygosity(Ho) was on average 0.458 ranging from 0.150 to 0.682. Polymorphism information content(PIC) values ranged from 0.625 to 0.865 with an average of 0.787. The analysis of molecular variance(AMOVA) highlighted a significant higher estimated variance within populations compared to among populations. Finally, the analysis of haplotypes by using cpSSR suggested a higher diversification in the population from Sila. Hierarchical clustering analysis grouped the genotypes into two major clusters, which agreed with the geographic origin of populations, and was confirmed by the Discriminant Analysis of Principal Components(DAPC). The first cluster included plants/population from Sila massif, while the second encompassed mostly plants/population sampled in Aspromonte massif. Finally, model-based clustering by STRUCTURE analysis also supported the presence of clear genetic structuring in the collection with two major populations(K=2) supported to PCoA analysis as well. Finally, our data indicated the Aspromonte population as a marginal forest with fragmented distribution suggesting different strategies of preservation than in Sila massif.     

Hybrids between olive flounder Paralichthys olivaceus and stone flounder Kareius bicoloratus: karyotype, allozyme and RAPD analyses

The hybrid between olive flounder Paralichthys olivaceus and stone flounder Kareius bicoloratus was produced by artificial insemination of olive flounder eggs with stone flounder Sperm. Sinistral and dextral are two types of hybrid progeny after metamorphosis. Karyotypes of both hybrid flounders are the same as those of the two parental species. Of the 22 loci examined from 12 allozymes,12 confirmed hybridization of the paternal and matemal loci in hybrids and no difference was found in allozyme patterns of sinistral and dextral hybrid fishes. RAPD patterns of these specimens were also studied with 38 primers selected from 104 tested. Among them, the PCR products of 30 primers showed hybridization of the paternal and maternal bands. Genetic variation between hybrids and their parental stocks was analyzed by RAPD using 10 of the above 38 primers. The average heterozygosity and genetic distance were calculated. The results suggested that the filial generation could inherit a little more genetic materials from paternal fish than that from maternal fish.