Genetic differentiation withinEriochoir sinensis (milne, edwards) revealed by allozyme and RAPD

We analyzed 17 allozymes, and 20 primers in order to detect the genetic differentiation between commercial populations (Changjiang River, Liaohe River) ofEriochoir sinensis. Ten allozymes (LDH, MDH, ME, IDH, EST, ALP, AAT, CTL, POD, SOD) showed 21 loci by vertically discontinuos buffer system polyacrylamide gel electrophoresis. RAPD profiles generated by 12 ten-base primers showed 63 loci. The percentage of polymorphic loci and the expected heterozygosity obtained by using allozyme analysis were lower than those obtained by RAPD. The index of similarity between these two populations were 0.955 and 0.932 as revealed by allozyme analysis and RAPD technology. There was gene flow between the above populations. Contribution No. 4025 from the Institute of Oceanology and Limnology, Chinese Academy of Sciences. Project 39470141 supported by NSFC. Work also supported by Climbing Program B (PDB 6-5-3) and No. 863 High-technology Program 863-819-01-05.     

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.     

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 maternal 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. Supported by National Natural Science Foundation of China (No. 30571445), National High-Technology Research and Development Program (863 Program, No. 2006AA10A404), and Project from the Ministry of Science and Technology of China (No. 2006DKA30470-017)     

The comparison between allozyme and RAPD makers for the population genetic structure analysis of scallop Chlamys farreri

1INTRODUCTION Since the first use in human genetic research in1966,allozyme has become the most popular genetic marker for population genetics studies for almost all groups of animals and plants(Lewontin and Hubby,1966;Murphy et al.,1996).In the past seve…     

Genetic variation of natural and cultured stocks of Paralichthys olivaceus by allozyme and RAPD

Population genetics of the left-eyed flounder, Paralichthys olivaceus, including natural and cultured stocks distributed in the coastal waters near Qingdao of eastern maritime China, was analyzed in allozyme and RAPD. The results showed that among total 29 gene loci of 15 isozymes, 9 and 7 were po- lymorphic in natural and cultured stocks, respectively. The status of genetic diversity in P. olivaceus is low in terms of polymorphic loci in chi-square test and genetic departure index of Hardy-Weinberg equi- librium. More alleles in IDHP, CAT, GDH and Ldh-C allozymes were found in the fish, which could be used as markers in assortive breeding and distinguishing stock, population or species evolution. Total 88 and 86 RAPD bands ranging from 200 to 2 500 bp were recognized individually in average of 7.8–8.0 bands per primer. The genetic diversity in cultured stock is lower than that in natural ones showing an ob- viously decreasing genetic divergence. Therefore, effective countermeasures must be taken to protect ge- netic resources of marine cultured fishes. The 2 markers have their own pros and cons. Combining the 2 markers to investigate the genetic variation of populations is suggested. The results provide basic data of this flounder and they are useful for studying genetic improvement and genetic resources of the fish.     

紫红笛鲷随机扩增多态性DNA及遗传多样性分析

采用RAPD技术分析了湛江近海紫红笛鲷 (LutjanusargentimaculatusForsk l)的遗传多样性。从 12 0个随机引物中筛选出了 14个引物。 14个引物共检测到 173个位点 ,其中多态位点比例 (P)为 6 8.79%,遗传距离 (D)为 0 .2 2 2 8,遗传多样性指数 (H)为 0 .190 4。结果表明 ,目前湛江近海的紫红笛鲷自然群体的遗传多样性仍然维持在良好水平 ,捕捞尚未对其造成明显影响。     

Genetic study of Kelp"901"strain

Based on DNA extraction and optimization of random amplified reaction (RAPD) to the gametophytes and sporophytes of Kelp “901” strain, genetic study on variation was conducted to its parents and offsprings of F₆, F₇, F₈, and F₉ generation. RAPD results have shown that among 30 selected primers for gametophytes, 297 loci ranging from 200 to 3 000 bp were obtained in the average of 9.9 loci for each primer. This indicated a high polymorphic rate with RAPD detection. UPGMA (unweighted pair-group method arithmetic average) analysis showed that each male and female gametophyte of a generation could be clustered into one pair separately. The genetic distances of the Kelp 901 generation were 0.3212–0.4767, and the maximum was between F₇ and F₈ (0.4767). Identity analysis showed that F₆ generation was more close to the female parent (0.6593), and F₇ generation was more close to the male parent (0.578 8). To the sporophytes study in 24 selected primers for RAPD amplification, 191 loci ranging from 230–2800 bp were obtained, in the average to each primer of 8.0 loci. The heterozygosity to six populations were male parent (0.2239), female parent (0.1072), F₆ (0.2164), F₇(0.2286), F₈(0.2296) and F₉(0.3172). The nearest genetic distance was 0.083 5(F₈, F₉). Total heterozygosity (H_T) of F₆, F₇, F₈ and F₉ generations was 0.3186, the average heterozygosity (H_S) for F₆, F₇, F₈ and F₉ generations was 0.2480, and deduced coefficient of population differentiation (G_st) was 22.2%. Six sequence characterized amplified regions (SCAR) were preliminary screened through RAPD analysis. It needed to be verified in detail as they are significant for molecular marker assistance in breeding and selectingLaminaria.     

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。     

长江下游鳊鱼遗传多样性的RAPD分析

应用RAPD技术对长江下游鳊鱼群体的遗传多样性进行分析,从40个随机引物中筛选出28个引物对鳊鱼24个个体的基因组DNA进行扩增,共检测到178个位点,分子片段在0.2～2.0kb之间,其中多态位点83个,占46.63%;个体间最大的遗传距离为0.1326,最小的遗传距离为0.0476,平均遗传距离为0.0885;群体的Nei基因多样性指数为0.2593,Shannon多样性指值为0.0986。与其他鱼类的遗传多样性研究结果相比,长江下游鳊鱼群体的遗传多样性处于中等水平。     

RAPD分析中最适样本量和位点数的研究

用25个引物,对40个样本扩增,研究了勒氏笛鲷随机扩增多态性DNA(RAPD)分析中的最适样本量和位点数。结果显示:在样本量达到20且位点数达到70个以上时,遗传距离趋于一个稳定的数值,即在RAPD分析中,样本数应达到20且位点数应达到70才能保证结果的可靠性。     

蜂巢石斑鱼随机扩增多态性DNA的初步研究

采用RAPD技术分析了湛江沿海野生蜂巢石斑鱼的遗传多样性 ,从 6个系列 1 2 0个引物筛选出1 6个引物 ,对 1 0尾蜂巢石斑鱼进行RAPD分析 ,1 6个引物共检测到 1 5 4个位点 ,其中 ,多态位点比例 (P)为 6 5 5 8% ,平均个体间遗传相似系数 (S)为 0 7887,平均个体间遗传距离 (D)为 0 2 1 1 3,遗传多样性指数 (H)为 0 1 776。研究结果表明 ,湛江沿海的野生蜂巢石斑鱼的遗传多样性较高。     

广东水稻纹枯病菌RAPD基因型与致病型关系

分析了广东水稻纹枯病菌(Rhizoctonia solani AG-1IA)致病型多样性和RAPD多态性之间的相互关系。结果表明,供试水稻纹枯病菌表现出较为丰富的致病型多样性(H=3.485)和遗传多样性,55个菌株接种9个水稻品种表现出39种致病类型。11条RAPD随机引物共扩增出91条谱带,其中多态性条带86条,多态率为94.51%。扩增出的22个选择性RAPD位点中,每个菌株都存在特有的RAPD分子指纹图谱。55个菌株在毒力数据、RAPD数据以及两者结合的数据聚类分析UPGMA树形图上形成几个不同的聚类类群,其中一个类群包括了大部分的菌株。毒力数据与RAPD分子数据的Jacarrd相似性距阵间的相关性系数比较低(r=0.0137),说明致病型与RAPD基因型性之间为相互独立的关系。     

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。     

Assessing the Germplasm of Laminaria （Phaeophyceae） with Random Amplified Polymorphic DNA （RAPD） Method

Eighteen gametophytes including L.japonica ,L.ochotensis and L.longissima,were verified with random amplified polymorphic DNA (RAPD) technique.Eighteen ten-base primers were chosen from 100 primers selected for final amplification test.Among the total of 205 bands amplified ,181(88.3%) were polymorphic ,The genetic distance among different strains ranged from 0.072 to 0.391,The dendrogram constructed by unweighted pair -group method with arithmetic(UPGMA) method showed that the female and male gamtophytes of the same cell lines could be grouped in pairs respectively,It indicated that RAPD analysis could be used not only to distin-gusih different strains of Laminariz ,but also to distinguish male and female gametophyte within the same cell lines,There is ambiguous systematic relationship if judged mereely by the present data It seems that the use of RAPD marker is limited to elucidstion of the phylogenetic relationship among the species of Laminaria.     

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。     

两种体色画眉笛雕的遗传多样性及分子标记

根据体侧纵带颜色差异,暂将画眉笛鲷(Lutjanus vitta)分为黄带画眉笛鲷和褐带画眉笛鲷,应用RAPD技术,选用22个引物对黄带画眉笛鲷与褐带画眉笛鲷进行遗传多样性及分子标记研究。结果表明,黄带画眉笛鲷与褐带画眉笛鲷的遗传距离为0.832 9,其多态位点比例(P)分别为66.99%和85.28%,遗传多样性指数(H)分别为0.161 7和0.289 3,种内遗传距离(D)分别为0.243 5和0.431 9,表明褐带画眉笛鲷的遗传多样性比黄带画眉笛鲷丰富。共得到OPP14-629bp和OPP14-674bp等11个特异性分子标记,可用于两种笛鲷的鉴定。     

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。     

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 differentiation of Japanese sardinella (Sardinella zunasi) populations in the Northwest Pacific revealed by ISSR analysis

Knowledge of population genetic structure plays an important role in fisheries management.In this research,In-ter-Simple-Sequence-Repeat(ISSR)markers were employed to evaluate the genetic structure of Japanese sardinella(Sardinella zun-asi)populations in the Northwest Pacific.Eighty seven individuals from 5 locations were screened using 4 highly polymorphic prim-ers.A total of 173 polymorphic loci were detected out of 191 loci amplified.Small but significant genetic differentiation was de-tected between the Chinese and Japanese populations by both AMOVA and pairwise FST analyses,which was further supported by cluster analysis.We consider that climate change during glaciations should be responsible for the genetic differentiation.Isolation by geographic distance among populations was observed,indicating that the distance might also lead to the genetic differentiation.However,no genetic structure was found within the populations off both the Chinese and Japanese coasts,indicating a high-level along-coast gene flow,which might result from ocean current transport and common ground for over-wintering.