细基江篱及坛紫菜太空诱变遗传变异的RAPD分析

细基江蓠(Gracilaria tsengiana)及坛紫菜(Porphyra haitanensis)经过卫星搭载送入太空,进行太空诱变试验,返回地面后采用RAPD技术对其基因组DNA多态性进行了检测.筛选出16个随机寡核苷酸引物进行扩增,其中细基江蓠检测出93个位点,多态位点31个,占33.3%;坛紫菜检测出84个位点,多态位点19个,占22.6%;分子量在150～2500 bp之间.遗传相似系数用Nei的计算方法进行计算,遗传距离则用Lynch的计算方法进行计算.实验数据表明:经过太空诱变的群体与正常对照组相比,从整个基因组角度来看,其遗传多样性差异并不明显,但是太空诱变使部分位点的基因产生了较为显著的变异.研究结果证实:太空诱变能够不定向地促进基因的变异,是水产新品种选育的途径之一.     

Genetic variation in different varieties of Siamese fighting fish using isoelectric focusing of sarcoplasmic proteins

Sarcoplasmic proteins of four colour varieties of Siamese fighting fish, Betta splendens Regan, were analysed by isoelectric focusing (IEF). Genetic polymorphisms were detected in six loci having isoelectric points: 4.92-5.00 (Sp-1), 5.33-5.48 (Sp-2), 5.90 (Sp-3), 6.31-6.35 (Sp-4), 6.58-6.70 (Sp-5) and 6.90 (Sp-6). For the Sp-1, Sp-2, Sp-4 and Sp-5 loci, there were two co-dominant alleles. At Sp-3 and Sp-6, one allele was dominant and the other null. Individuals within each locus were classified into phenotypes A, AB and B. Only homozygotes were found at Sp-2. Allele frequencies were calculated for each locus and tested for accordance with the Hardy-Weinberg equilibrium. Observed values for each phenotype at Sp-1, Sp-3, Sp-4, Sp-5 and Sp-6 agreed well with the expected ones. At Sp-2, genotypic frequencies for each variety deviated markedly (p < 0.01) from the equilibrium. When tested for homogeneity, no significant differences (p 0.95) in gene frequencies were found among the varieties except at Sp-2. Band sharing index (BSI), computed using the BIO-GENE v5.04 program, revealed high intravariety genetic similarities (0.701-0.740) and slightly above-average intervariety values (0.550-0.583). In this study, we estimated the levels of genetic variability in B. splendens, and proposed a genetic model for each polymorphic locus.     

漠斑牙鲆引进群体子一代遗传多样性的RAPD分析

运用随机扩增多态性DNA（RAPD）技术对引进群体漠斑牙鲆Paralichthys lethostigma 子一代48个个体的遗传多样性进行了分析,筛选的40个10bp的随机引物中,25个引物产生了稳定性好,可重复性强的谱带。共检测到154个位点,其中多态位点有122个,多态位点的比例为79．22％,有效等位基因数为1．5629±0．3631,Nei＇s基因多样性指数为0．3184±0．1843,Shannon信息指数是0．4651±0．2574。结果表明,漠斑牙鲆处于较高的遗传多样性水平。     

平胸龟遗传多样性的RAPD分析

应用RAPD技术对平胸龟(Platysternon megacephalum Gay)的遗传多样性进行了分析,用27个随机引物对福建寿宁平胸龟21个个体的基因组DNA进行了PCR扩增,共扩增出5187条DNA片段,平均每个个体扩增出247条带。在检测到的247条带中,多态性条带为141条,多态性条带百分比为55.97%(25.0%~88.89%),条带大小在100bp~2000bp之间,21个个体间遗传距离为0.0972~0.3198,平均遗传距离为0.1750±0.0387,表明平胸龟具有较高的遗传多样性水平。采用类平均聚类法(NJTREE)构建了21个个体相互关系的分子聚类图,表明该地区平胸龟并没有形成种群的分化。     

应用RAPD标记对东方(鱼屯)属进行种类鉴别及其聚类分析

利用RAPD技术对红鳍东方、假睛东方、暗纹东方和从日本引进的红鳍东方4个种进行了遗传标记鉴别研究.在事先优化的条件下,20个随机引物共扩增出134条谱带清晰、重复性高的DNA片段,其中多态性片段77条.结果证实每一种均有其特异性扩增图谱,可作为种鉴定的依据;用UPGMA和NJ等方法进行聚类,构建的系统树与借助形态学和生化特征进行的传统分类结果一致.研究表明,RAPD作为一种新的分子标记,在海洋动物遗传鉴定方面具有巨大的潜力.     

应用RAPD标记对东方鲀属进行种类鉴别及其聚类分析

利用RAPD技术对红鳍东方鲀、假晴东方鲀、暗纹东方鲀和从日本引进的红鳍东方鲀4个种进行了遗传标记鉴别研究。在事先优化的条件下，20个随机引物共扩增出134条谱带清晰、重复性高的DNA片段，其中多态性片段77条。结果证实每一种均有其特异性扩增图谱，可作为种鉴定的依据；用UPGMA和NJ等方法进行聚类，构建的系统树与借助形态学和生化特征进行的传统分类结果一致。研究表明，RAPD作为一种新的分子标记，在海洋动物遗传鉴定方面具有巨大的潜力。     

7种珍珠贝RAPD鉴别标记的初步研究

采用RAPD分子标记技术对珠母贝属的大珠母贝、珠母贝、黑珠母贝、白珠母贝、合浦珠母贝、长耳珠母贝和珍珠贝属的企鹅珍珠贝的基因组DNA的特异性遗传标记进行分析。从21个OPM和S系列中筛选出4个引物,共扩增出57个位点,每条引物平均产生14·3个位点。扩增片段大小在250~2000bp间,平均每种贝每条引物产生4·9条带。其中引物S10对7种珍珠贝的RAPD产物呈现出物种的特异性,可同时将7种珍珠贝分开,其余引物可以将2种或2种以上的珍珠贝区别开来。引物S10可以作为种间鉴定的标记。     

用同工酶和RAPD技术分析黑斑口虾蛄的遗传多样性

采用同工酶技术和RAPD技术对宁波黑斑口虾蛄的遗传多样性进行了分析,采用聚丙烯酰胺凝胶电泳方法(PAGE)对LDH、MDH、MEP、EST、ADH、GDH、GLDH、AMY、POD、SDH等10种同工酶进行了群体生化遗传分析,共检测了29个基因座,其中11个基因座表现为多态,其多态位点比例为37.93%,平均杂合度为0.1470;采用RAPD技术对其遗传多样性进行检测,用24个随机引物共检测出129个位点,其中多态位点为102个,占79.06%,个体间遗传相似系数为0.3106,遗传变异度为0.6894,表明黑斑口虾蛄的遗传多样性状况良好。     

罗氏沼虾浙江养殖群体与缅甸自然群体遗传差异的RAPD分析

利用随机扩增多态DNA(RAPD)技术，对浙江省罗氏沼虾养殖群体和新引进的缅甸自然群体的遗传差异进行了比较分析，以期从分子水平了解罗氏沼虾的种群遗传多样性背景及与引种的关系。采用经筛选的22个10bp的随机引物对罗氏沼虾两群体各20尾进行群体RAPD分析。22个引物共检测到139个位点。养殖群体的多态位点比例为30．22％，群体平均杂合度为0．2646，Shannon多样性指数为0．0780，群体内各个体之间的遗传共享度为0．9353，遗传距离为0．0647；自然群体的多态位点比例为33．81％，群体的平均杂合度和Shannon多样性指数分别为0．2888和0．0940，群体内各个体之间的遗传共享度为0．9201，遗传距离为0．0799；两群体之间的遗传距离为0．1845。自然群体的遗传多样性水平比养殖群体要高。利用随机引物S9和S52，在罗氏沼虾两群体间扩增出2条稳定、明显的群体间特异性标记带。     

中国太湖和荷兰的中华绒螯蟹随机扩增多态性DNA分析

用随机扩增多态DNA(RAPD)技术对中华绒螯蟹两个群体(荷兰盐城湖和中国太湖)进行了检测,从40个随机引物中筛选出14个对两群体进行扩增分析。结果为:14个引物共检测到100条清晰且重复性好的条带,分子量在200~2000bp之间,太湖和盐城湖两群体的多态位点比例、Shannon多样性指数、Ne i基因多样性指数分别为:46%和44%、0.2195和0.1976、0.1446和0.1270,两群体的遗传距离为0.013。据此结果推断,中华绒螯蟹群体遗传多样性不高,种内群体间的遗传变异较低,其遗传资源亟待保护,同时验证了荷兰的中华绒螯蟹是从中国引进繁衍的推论。     

RAPD fingerprinting of the eel-loaches Pangio filinaris and Pangio piperata: preliminary evaluation

Random amplified polymorphic DNA (RAPD) analysis was applied to two morphologically similar species of the increasingly popular ornamental eel‐loaches, Pangio filinaris and P. piperata, collected from Kemat River, Terengganu and Padang Sanai River, Kedah, West Malaysia. Forty primers were tested in a preliminary screening. Of these, five (OPA‐03, OPA‐09, OPA‐11, OPA‐13 and OPA‐18) were chosen for their ability to provide consistent amplification. RAPD analysis differentiated the two species, with each showing different RAPD patterns for each primer used. The five primers generated a total of 82 scorable loci with up to 83% and 60% polymorphism in P. piperata and P. filinaris respectively. The RAPD banding patterns of these two species ranged from 2 to 14 fragments, with a size range of 250–2000 bp. Genetic similarity within species ranged from 0.610 to 0.985 (mean 0.799±0.14) for P. piperata and from 0.824 to 0.934 (mean 0.885±0.04) for P. filinaris respectively. The interspecies genetic similarity ranged from 0.386 to 0.486 with an average value of 0.434±0.040. Our study revealed RAPDs as useful genetic markers for the taxonomic clarification and assessment of genetic variability of the eel‐loaches.     

Genetic diversity and species identification of Argulus parasites collected from major aquaculture regions of India using RAPD‐PCR

Argulus is one of the most important fish parasites that cause heavy economic loss to aquaculture industry. The present investigation was undertaken to study the genetic diversity of the Argulus sp. collected from 13 locations representing major aquaculture zones in India by RAPD analysis and to develop species‐specific markers. Thirteen random decamer primers were used to amplify DNA fragments from three individual parasites of each location. Of the 172 bands scored by the primers, 168 were polymorphic. The per cent polymorphic loci and gene diversity values varied within a range of 8.14–43.02 and 0.0342–0.1727 respectively. Nei's genetic similarity between populations across all the primers ranged from 0.363 to 0.969. The dendrogram based on Nei's genetic distance showed two clusters; Bangalore and Mandi populations forming one cluster, and the rest in another cluster. The clusters also revealed strong correlation with the species identified as A. japonicus and A. siamensis respectively by morphological method. The study thus indicated A. siamensis as the major prevalent species in carp culture farms in India. Species‐specific primers were designed from unique sequences cloned from RAPD fragments that could able to identify A. siamensis and A. japonicus separately.     

Population genetic structure and variability of Pacific herring <Emphasis Type="Italic">Clupea pallasii</Emphasis> in the stocking area along the Pacific coast of northern Japan

ABSTRACT:   The genetic diversity of wild and hatchery-released Pacific herring Clupea pallasii collected from three brackish lakes and two bays in Honshu and Hokkaido, Japan was examined with five microsatellite loci. All loci showed high genetic variability with expected heterozygosities ranging from 0.815 to 0.945. Significant differences in genotypic and allelic distributions were detected among all locations except for between the two bays in Honshu Island. Pairwise population analysis based on the F _ST values showed close genetic relationships among the locations in Hokkaido Island, and the hierarchical analyses of molecular variance showed significant genetic difference between the two islands. Those results suggest the existence of subpopulations due to natal homing. In addition, stocked fish showed as much genetic diversity as the wild fish. The pairwise population analyses also showed close relationships between the hatchery fish and the wild fish in respective stocking areas, showing that no effects of stocking programs on genetic diversity of wild populations were detected.     

~(60)Co-γ射线对长毛明对虾诱变后子代幼体遗传变异的RAPD分析

本文首次使用60Co-γ射线对长毛明对虾进行诱变,将性腺成熟的长毛明对虾(Fenneropenaeus penicillatus)分为四组进行不同剂量的60Co-γ射线照射,采用RAPD技术对诱变子代的无节幼体基因组DNA多态性进行了检测。遗传相似系数用Nei的计算方法进行计算,遗传距离则用Lynch的计算方法进行计算。从20个随机寡核苷酸引物共检测154个位点,其中多态位点103个,占66.9%。单个引物获得的标记为4~11个,平均7.7个,分子量在100~2500bp之间。计算得到的遗传相似系数变化范围在0.7995~0.8696之间,相应的遗传距离变化范围则在0.1397~0.2288之间。实验表明:诱变子代与正常对照组相比,遗传多样性水平较高,诱变产生了较为显著的变异。     

Wild stock structure of <Emphasis Type="Italic">Girella punctata</Emphasis> in Japan revealed shallow genetic differentiation but subtle substructure in subsidiary distributions

Twelve populations of Girella punctata, from widespread locations of the species’ range in Japan and Korea, were screened for sequence variability within the mitochondrial DNA (mtDNA) control region (n = 128) and at five polymorphic microsatellite loci (n = 547) to determine the genetic structure maintaining population integrity. mtDNA variability of 132 variable sites within a 334-bp region reveals shallow genetic differentiation across populations. The weak differentiation of G. punctata was partly supported by the screening of five polymorphic microsatellite loci. However, hierarchical analysis of molecular variance and principal component analysis on the basis of allele frequencies in microsatellite loci extracted a subtle substructure in a subsidiary population and in near-subsidiary populations in the semi-enclosed Seto Inland Sea.     

镜鲤与建鲤生长性状共享 QTL 标记及优势基因型

本研究以1个镜鲤(Cyprinus carpio L.)全同胞家系(190个个体)构建的微卫星遗传图谱(992个标记)为基础,从体重、体长、体高和体厚的QTL区间内发掘了54个标记,其与性状具有显著相关性,进而通过对不同基因型性状间的比较,筛选出83个优势基因型。在此基础上,用54个镜鲤QTL标记分析了建鲤(Cyprinus carpio var.jian)作图群体,其中40个标记在建鲤中表现出多态性,比例为74.07%;相关性分析结果显示,其中22个标记与建鲤家系的体重、体长、体高或体厚性状具有显著相关性(P0.05),占多态标记的55.00%;镜鲤与建鲤共享的22个QTL标记中,18个标记与至少1个相同的性状具有显著相关性(P0.05),从中筛选出建鲤性状具有优势的基因型30个,可用于指导建鲤的选育。品种间共享QTL的发掘能够扩展QTL标记的使用空间,减少新品种重新构建图谱进行QTL标记定位的工作量和成本。     

中国对虾5个地理群体的RAPD分析

采用随机扩增多态DNA(RAPD)技术对取自辽东湾、渤海湾、海州湾、乳山湾及海洋岛5个群体的中国对虾共95个个体的遗传多样性进行分析。14个随机引物共检测出103个位点(200～2500bp),各群体的多态位点比例在31．07％～34．95％之间。遗传距离显示中国对虾群体间有一定遗传分化,其中辽东湾和乳山湾群体问的遗传距离最大(0．0742),而辽东湾和渤海湾群体间的遗传距离最小(为0．0243),群体间的遗传分化系数为0．2083。整体来看,中国对虾的遗传变异水平较低,遗传多态度为0．1621。用UPGMA对5个群体进行聚类分析,构建谱系关系图。     

江苏及安徽地区十个中华绒螯蟹成蟹群体的RAPD分析

采用随机扩增多态DNA(RAPD)技术对取自江苏及安徽地区的10个群体的中华绒螯蟹共64个个体进行了遗传多样性分析.从24个随机引物中筛选出15个扩增重复性好、条带清晰、特异性强的引物,共检测出88个位点(100～2000 bp),各群体的多态位点比例为3.45%～26.19%,遗传多态度为0.0141～0.1036,说明该蟹类基因组DNA多态度较为贫乏,遗传变异水平较低;各群体间的遗传距离为0.0237～0.2466,遗传相似度为0.7815～0.9766,该蟹类群体之间发生了一定程度的分化;分化系数为0.4283～0.6632,Nm为0.2539～0.6674,表明该蟹类群体之间发生了不同程度的遗传变异和遗传交换.     

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.     

Validation of 12 species-specific,tetrasomic microsatellite loci from the Russian sturgeon, <Emphasis Type="Italic">Acipenser gueldenstaedtii</Emphasis>, for genetic broodstock management

The Russian sturgeon, Acipenser gueldenstaedtii, is a critically endangered fish species. Hatcheries are operated in several countries within its natural range to produce stocking material for release into the wild and also for aquaculture purposes (caviar and meat production). An appropriate genetic broodstock management (plan or strategy) is required to avoid negative effects, e.g., admixture and hybridization of genetically differing stocks or loss of genetic variability due to inbreeding and genetic drift. Therefore, 11 tetrasomic microsatellite loci were newly isolated from the Russian sturgeon genome and arranged together with an already known locus into four multiplex PCR sets. These microsatellites were used to characterize three groups of hatchery juveniles from Germany (aquaculture production), Turkey, and Romania (production of stocking material) as well as a group of wild-caught adults from the Danube River, Romania. Based on the variability within groups, measured by the mean number of alleles per locus and expected heterozygosity, and the differentiation between groups, measured by Nei’s G_ST and genetic distance D, the ability of the 12 loci to detect unwanted reductions in genetic variability within hatchery juveniles and to differentiate between groups could be demonstrated. This set of loci can also be used to identify those pairs of spawners that transmit the highest possible genetic variability to the next generation.