近江牡蛎3个种群生化遗传变异的研究

用同工酶电泳方法研究广东省官渡、阳江和汕尾的近江牡蛎（Ostea rivularis）的遗传变异。分析了11种同工酶20个的位点，其中出现13个位点显示为多态。结果表明官渡、阳江、汕尾种群的多态位点比例分别为0．200、0．200、0．500。官渡和阳江种群间，官渡和汕尾种群间以及阳江和汕尾种群间的遗传距离（D）分别为0．0451，0．1125,0．1088。     

近江牡蛎两个野生种群的遗传多样性分析

采用RAPD分子标记和rDNA-ITS1序列分析了近江牡蛎Crassostrea rivularis湛江官渡和阳江程村野生种群的遗传多样性。12个RAPD引物共扩增出8838条片段,157个位点,平均每个引物可产生13个位点,片段长度在200～2200bp之间。湛江种群和阳江种群的多态位点比例分别为89.62%和89.57%,遗传多样性指数分别为0.4170和0.4334。种群间平均遗传距离为0.0327,平均遗传相似性为0.9681,平均遗传分化系数为0.0437。得到近江牡蛎18S、5.8S部分序列和ITS1全部序列,其中ITS1序列片段长度为478～485bp,共有11个变异位点,两个为转换(A/G),其他为插入/缺失(A/-、T/-)。湛江和阳江种群各获得8个单倍型,其中有一个单倍型为两个种群共享。两个种群的CG碱基平均含量较高,分别为58.29%和58.41%。种群间的遗传分化系数0.0254。结果说明,近江牡蛎湛江种群和阳江种群间具有较高的遗传多样性和较低的遗传分化。     

褐菖鲉同工酶的组织特异性及群体遗传结构

用聚丙烯酰胺垂直板不连续凝胶电泳法对湛江海域褐菖鲉(Sebastiscus marmoratus)心、肝、肾、性腺、肌肉、鳃、眼、脑等8种组织的12种同工酶(ADH、a-AMY、CAT、EST、GDH、LDH、MDH、ME、POD、PPO、SDH、SOD)进行分析。结果表明,12种同工酶均有明显的组织特异性。经筛选,对肝脏和眼的8种同工酶(ADH、EST、LDH、MDH、ME、POD、SDH、SOD)进行群体遗传结构分析,共记录了18个基因位点,其中呈多态性的基因位点有5个(Adh-1、Est-2、m-Pod-1、s-Pod-1、s-Sod-1),多态位点比例(P)为27.78%,平均有效等位基因数(Ae)为1.238 8,平均观测杂合度(Ho)值为0.1401。     

运用RAPD技术分析3种罗非鱼的遗传多样性

应用RAPD技术,从7个系列140种随机引物中筛选出15个,对尼罗罗非鱼(Oreochromis niloticus)、奥利亚罗非鱼(O.aureus)和星洲银罗非鱼(O.sp.)进行遗传多样性分析。结果表明,尼罗罗非鱼、奥利亚罗非鱼、星洲银罗非鱼的多态位点比例(P)分别为54.7%、44.4%和56.0%,种内个体间遗传距离分别为0.145 0、0.115 6和0.141 3,种内遗传相似系数分别为0.855 0、0.884 6和0.858 7;种间遗传距离的聚类分析表明,星洲银罗非鱼与尼罗罗非鱼亲缘关系较近,一定程度上验证了星洲银罗非鱼的分类地位。     

翡翠贻贝三个野生种群形态性状的差异分析

运用差异系数检验、主成分分析、判别分析和聚类分析等4种多元统计分析方法,采用5个比例性状,比较研究了广西北海、广东湛江和汕尾的3个翡翠贻贝野生种群的形态性状差异。结果表明:(1)3个种群间5个比例性状的差异系数在0.024～1.134之间,种群间尚未达到亚种差异水平;(2)主成分分析构建了3个主成分,PC1、PC2和PC3贡献率分别为40.134%、27.956%和20.614%,累计贡献率为88.704%;(3)利用逐步判别法选择3个贡献率较大的性状进行判别分析,建立了3种群判别函数,判别准确率为P1为73.0%～88.0%,P2为74.6%～86.8%,综合判别率为79.7%;(4)聚类分析显示,北海种群与汕尾种群形态较为接近,湛江种群趋异程度最大。各种分析方法从不同角度反映了群体间的形态学差异,这种差异是环境和遗传共同作用的结果。     

运用RAPD技术对笛鲷属3种鱼的群体遗传学分析

应用RAPD技术,从9个系列180种随机引物中筛选出16种,对笛(鱼周)属的画眉笛(鱼周)(Lutja-nus vitta)、金焰笛(鱼周)(Lutjanus fulviflamma)、金带笛(鱼周)(Lutjanus vaigiensis)进行种群内及种群间遗传学分析,并利用UPGMA确定了它们之间的亲缘关系。结果表明,三种鱼的种内遗传多样性指数(H)分别为,画眉笛(鱼周)0.1949,金带笛(鱼周)0.1107,金焰笛(鱼周)0.1673;三者的多态位点比例(P)分别为69.4％、47.8％和59.0％;金带笛(鱼周)具有较低的遗传多样性,且全带笛(鱼周)与金焰笛(鱼周)有较近的亲缘关系。三种鱼共检出10个可作为种的特异性鉴定的条带,可用于种质鉴定。     

军曹鱼群体遗传结构的AFLP分析

筛选出10对扩增片段长度多态分析技术(Amplified fragment length polymorphism,AFLP)的选择性扩增引物(E-aga/M-cgt、E-aga/M-cga、E-agc/M-cga、E-agg/M-cga、E-act/M-cgc、E-aag/M-cgc、E-aca/M-cga、E-aac/M-cgc、E-aac/M-cac和E-aac/M-cat),分析了4个军曹鱼(Rachycentron canadum)全人工繁育群体海南1(HN1)、海南2(HN2)、湛江(ZJ)和流沙湾(LS)的遗传结构。结果表明:总体多态位点比例(83.9%)、分化指数(0.427 8)和Shannon信息指数(0.614 9)均处于较高水平,遗传多样性丰富;主坐标分析和邻位连接树中LS-HN2支与ZJ-HN1支明显分离,各群体内多样性水平存在差异(遗传多样性大小依次为HN1、HN2、LS、ZJ),但总体基因流仍然较大(Nm=10.327),分子方差分析(AMOVA)获得的群内方差(53%)也略大于群间(47%)。     

似刺鳊鮈发育过程中同工酶变化及组织分布特征分析

运用高pH不连续聚丙烯酰胺凝胶(PAGE)电泳技术分析了似刺鳊鮈胚胎、仔鱼发育过程中5种同工酶变化,并对成鱼的9种组织同工酶分布特征进行了检测。结果发现:除醇脱氢酶(ADH)外,其他4种酶(EST、LDH、MDH、SOD)在发育过程中均被检出,且呈现明显的阶段性特征;在成鱼的4种同工酶也存在明显的组织特异性。4种酶共检测到9个基因位点。     

运用RAPD技术对笛鲷属3种鱼的群体遗传学分析

应用RAPD技术，从9个系列180种随机引物中筛选出16种，对笛鲷属的画眉笛鲷(Lutja-nus vitta）、金焰笛鲷(Lutjanus fulviflamma)_，金带笛鲷(Lutjanus vaigiensis)进行种群内及种群间遗传学分析，并利用UPGMA确定了它们之间的亲缘关系。结果表明，三种鱼的种内遗传多样性指数(H)分别为，画眉笛鲷0．1949，金带笛鲷0．1107，金焰笛鲷0．1673；三者的多态位点比例(P)分别为69．4％、47．8％和59．0％；金带笛鲷具有较低的遗传多样性，且金带笛鲷与金焰笛鲷有较近的亲缘关系。三种鱼共检出10个可作为种的特异性鉴定的条带，可用于种质鉴定。     

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。     

GENETIC DIVERSITY OF THE WILD AND REARED PSEUDOSCIAENA CROCEA

ＩＮＴＲＯＤＵＣＴＩＯＮＴｈｅｌａｒｇｅｙｅｌｌｏｗｃｒｏａｋｅｒＰｓｅｕｄｏｓｃｉａｅｎａｃｒｏｃｅａｏｎｃｅｗａｓｏｎｅｏｆｔｈｅｆｏｕｒｉｍｐｏｒｔａｎｔｍａｒｉｎｅｆｉｓｈ ｅｒｙｏｂｊｅｃｔｓｉｎＣｈｉｎａ (Ｈｏｎｇｅｔａｌ.,1 983;ＨｕａｎｇａｎｄＣａｒｌ,1 983;ＣｈｅｎａｎｄＧｏｎｇ ,1 983) .ＴｈｅｐｒｏｄｕｃｔｉｏｎｏｆｔｈｅｗｉｌｄＰ .ｃｒｏｃｅａｉｎ 1 974ｒｅａｃｈｅｄ 1 971 78ｍｅｔｒｉｃｔｏｎｓ (ＭＴ)ｉｎＣｈｉｎａ (Ｌｉ,1 998) .Ｂｅｃａｕｓｅｏｆｏｖｅｒｆｉｓｈｉｎｇ ,ｔｈ…     

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.     

东、黄海鲐鱼群体遗传差异的RAPD分析

利用随机扩增多态DNA(RAPD)技术分别分析了五岛西部日本鲐、东海西部日本鲐和澳洲鲐三群体的样本,共产生226个分子标记,其中143个为多态性标记,多态百分率达到63.3%,每条引物产生的平均标记数为7.29。五岛西部日本鲐、东海西部日本鲐和澳洲鲐群体内遗传相似度分别为0.949、0.877和0.936,而群体间遗传距离依次为:五岛西部日本鲐-东海西部日本鲐(0.2357)<东海西部日本鲐-澳洲鲐(0.3030)<五岛西部日本鲐-澳洲鲐(0.3528),五岛西部日本鲐与东海西部日本鲐个体在UPGMA系统树独立聚类为两个类群,初步支持东、黄海的日本鲐存在东海西部和五岛西部的两个种群的观点。Shannon多样性指数表明,其遗传多样性大小依次为东海西部日本鲐、澳洲鲐、五岛西部日本鲐。     

Genetic analysis of four populations of redlip mullet (Chelon haematocheilus) collected in China seas

Horizontal starch gel electrophoresis was used to investigate genetic structures of four populations (Dandong, Rizhao, Ningbo and Guangzhou) of redlip mullet (Chelon haematocheilus). Fourteen loci of ten enzymes (G3PDH, IDHP, LAP, LDH, MDH, MPI, PGDH, PGM, SDH, and SOD) were identified. The proportions of polymorphic loci varied from 0.07 to 0.14. The average observed and expected heterozygosities ranged from 0.02 to 0.05 and from 0.02 to 0.04 respectively among the 4 populations. The average number of efficient alleles of each locus varied from 1.02 to 1.06. The genetic distance among populations was from 0.0004 to 0.0021. The genetic distances between Dandong, Rizhao and Ningbo populations were small, while those between Guangzhou population and other three populations were relatively high. The result indicated the possible divergence of redlip mullet between the South China Sea and the other sea areas of China due to geographic isolation.     

Genetic Analysis of Four Populations of Redlip Mullet(Chelon haematocheilus) Collected in China Seas

1 Introduction The redlip mullet, (Chelon haematocheilus) (Tem-minck et Schlegel, 1845), belongs to the family Mugili-dae. It is mainly distributed in the coastal areas of Korea, Japan and China (Wang et al., 2000). Because of the good adaptation to envir…     

南海海域4种笛鲷的微卫星DNA分析

采用聚丙烯酰胺凝胶电泳技术,从20对西大西洋笛鲷(Lutjanus campechanus)的微卫星引物中筛选适用于红鳍笛鲷(L.erythopterus)、勒氏笛鲷(L.russellii)、紫红笛鲷(L.argentimaculatus)和约氏笛鲷(L.johnii)基因组分析的微卫星引物,分析4种笛鲷的遗传多样性及系统发生。经反应条件的优化,从20对引物中筛选出17对可稳定扩增出特异片段的引物。通过测序证实扩增产物含微卫星位点后,以该17对引物对4个种的群体进行遗传多样性分析。其中16对可在约氏笛鲷、勒氏笛鲷、紫红笛鲷基因组中扩增出重复性好的特异性条带,分别有65%、65%、60%呈现出种内多态;15对可在红鳍笛鲷中扩增出重复性好的特异性条带,并且全部呈现出种内多态。四种笛鲷中,红鳍笛鲷的多态性最高,高度多态基因座占检测座位的66.67%;勒氏笛鲷的多态性最低,低度多态基因座占43.75%。获得3个种间特异性分子标记。用DS和DA两种方法计算4种笛鲷的遗传距离,构建了系统发生树。     

Development and Validation of 89 Novel Expressed Sequence Tag-Derived Microsatellite Markers in Blood Clam,Tegillarca granosa

Blood clam, Tegillarca granosa, is an important shellfish in Chinese mariculture industry. Investigative research in this species, such as genetic linkage mapping, requires a large panel of molecular markers. In present study, a total of 89 polymorphic microsatellite markers were developed in T. granosa using the sequence database of Life Sciences Technology 454 next generation sequencing technology. All 89 loci were characterized in 20 individual clams from a natural population inhabiting Yueqing Gulf, Zhejiang Province, China. The number of alleles per polymorphic locus varied between 2 and 15, while the observed heterozygosity, expected heterozygosity and polymorphic information content varied between 0.000 and 1.000, 0.102 and 0.921, and 0.048 and 0.886, respectively. Of the 89 loci identified, 32 loci deviated significantly from Hardy-Weinberg equilibrium following Bonferroni correction. Thirty nine markers, which were shown to be polymorphic in a full-sibling family, were tested in Mendelian segregations. As expected, 32 loci were co-dominantly segregated in a Mendelian fashion. These novel developed microsatellite markers represent useful research tools for investigation of population genetic structure and genetic diversity in this species.     

Development and validation of 89 novel expressed sequence tag-derived microsatellite markers in blood clam, <Emphasis Type="Italic">Tegillarca granosa</Emphasis>

Blood clam, Tegillarca granosa, is an important shellfish in Chinese mariculture industry. Investigative research in this species, such as genetic linkage mapping, requires a large panel of molecular markers. In present study, a total of 89 polymorphic microsatellite markers were developed in T. granosa using the sequence database of Life Sciences Technology 454 next generation sequencing technology. All 89 loci were characterized in 20 individual clams from a natural population inhabiting Yueqing Gulf, Zhejiang Province, China. The number of alleles per polymorphic locus varied between 2 and 15, while the observed heterozygosity, expected heterozygosity and polymorphic information content varied between 0.000 and 1.000, 0.102 and 0.921, and 0.048 and 0.886, respectively. Of the 89 loci identified, 32 loci deviated significantly from Hardy-Weinberg equilibrium following Bonferroni correction. Thirty nine markers, which were shown to be polymorphic in a full-sibling family, were tested in Mendelian segregations. As expected, 32 loci were co-dominantly segregated in a Mendelian fashion. These novel developed microsatellite markers represent useful research tools for investigation of population genetic structure and genetic diversity in this species.     

Isozyme analysis of spotted halibut Verasper variegatus temminck et schlegel

To investigate the tissue-specificities of isozymes and the genetic structure of wild spotted halibut (Verasper variegates) population, horizontal starch gel electrophoresis was performed on 45 individuals collected in part of the Yellow Sea. The performances of 17 isozymes in 8 kinds of tissues or organs were screened preliminarily in a TC-7.0 buffer system. The results showed that the screened isozymes displayed remarkable tissue-specificities. Finally, 14 enzymes (AAT, ADH, EST, GPI, G3PDH, IDHP, LAP, LDH, MDH, MPI, PGDH, PGM, SDH and SOD) and 4 kinds of tissues (eye, skeleton muscle, liver and heart) were selected for genetic analysis. Fourteen isozymes are encoded by 20 loci, and 9 of them are polymorphic. The polymorphic loci are AAT-1*. GPI-2* G3PDH*, IDHP-1*, LDH*, MPI*, PGM-1*. PGM-2* and SDH*, and the proportion of polymorphic loci is 0.4500 (P0.99). The mean values of observed and expected heterozygosities are 0.027 8 and 0.026 5, respectively and the average effective number of alleles is 1.067 5.     

Isozyme Analysis of Spotted Halibut Verasper variegatus Temminck et Schlegel

To investigate the tissue-specificities of isozymes and the genetic structure of wild spotted halibut ( Verasper variegatus) population, horizontal starch gel electrophoresis was performed on 45 individuals collected in part of the Yellow Sea. The performances of 17 isozymes in 8 kinds of tissues or organs were screened preliminarily in a TC-7.0 buffer system. The results showed that the screened isozymes displayed remarkable tissue-specificities. Finally, 14 enzymes (AAT, ADH, EST, GPI, G3PDH, IDHP, LAP, LDH, MDH, MPI, PGDH, PGM, SDH and SOD) and 4 kinds of tissues (eye, skeleton muscle, liver and heart) were selected for genetic analysis. Fourteen isozymes are encoded by 20 loci, and 9 of them are poly-morphic. The polymorphic loci are AAT-1*, GPI-2*, G3PDH*, IDHP-1*, LDH*, MPI*, PGM-1*, PGM-2* and SDH*, and the proportion of polymorphic loci is 0.450 0 (P0.99). The mean values of observed and expected heterozygosities are 0.027 8 and 0.026 5, respectively and the average effective number of alleles is 1.067 5.