中国南方沿海13种石斑鱼类的分子系统进化关系分析

石斑鱼类(Groupers)是世界性海洋经济鱼类,也是中国南方沿海省份重要的海水增养殖对象,其种类繁多,由于缺乏明显的种间外部形态区别特征,石斑鱼的分类一直是鱼类系统分类学的一个难题,存在着较多的争议和混淆。为了从分子水平揭示石斑鱼类的系统进化关系,应用线粒体细胞色素b基因(Cyt b)及两个核糖体RNA基因(16S和12S)序列联合构建系统进化树对中国近海石斑鱼亚科3属13种进行种类鉴定及亲缘关系分析,探讨了石斑鱼类的分子系统进化。结果表明:①线粒体基因序列在石斑鱼类种间存在较大的变异,种内变异极小,是进行石斑鱼类种类鉴定较为适宜的分子标记;②所研究的3属13种石斑鱼类中,石斑鱼属与九棘鲈属的亲缘关系较近,与鳃棘鲈属的较远;③赤点石斑鱼和青石斑鱼、蜂巢石斑鱼与鲑点石斑鱼、斜带石斑鱼与棕斑石斑鱼两两间具有较近的亲缘关系。     

石斑鱼肌肉氨基酸组成的初步研究

本文测定了花点石斑鱼和青石斑鱼肌肉中18种氨基酸的含量。测定结果表明:石斑鱼肌肉中氨基酸含量较高,花点石斑鱼肌肉中氨基酸总含量为764.98mg/g,青石斑鱼的为743.78mg/g,这两种石斑鱼肌肉的氨基酸总舍量中,人体的8种必需氨基酸含量之和分别为40.87%和41,99%;谷氨酸、天门冬氨酸,精氨酸、丙氨酸和甘氨酸等5种氨基酸系的鲜味物质的含量之和分别为45.03%和44.37%.石斑鱼肌肉所含必需氨基酸的比值与人体所需氨基酸的组成模式比较接近.所以,对人体来说,石斑鱼肌肉是一种营养价值较高的鲜美的蛋白源.最后,本文提出了石斑鱼必需氨基酸的组成模式。     

Differentiation of Epinephelus moara from E. bruneus by improved nest-tetra-primer-specific PCR

Epinephelus moara and E. bruneus are closely related species in the genus Epinephelus (Perciformes, Serranidae). Their morphological similarity, changing color pattern at different stages and living conditions make them difficult to be differentiated. To identify these two species, an improved nest-tetra-primer-specific PCR assay was developed. Three specific molecular markers, the control region NC1 (394 bp), species-specific internal region ND2-M (268 bp) and ND2-B (122 bp), were identified in the mitochondrial ND2 gene from these two grouper species. Five markers were also discovered in the ITS1 regions of their nuclear ribosomal DNA, which were the control regions NC2 (588 bp) and NC3 (563 bp), and species-specific internal regions rDNA-M (426 bp), ITS1-M (488 bp) and ITS1-B (304 bp). This method provided a highly specific, precise, reliable and rapid molecular marker technique to discriminate between the two grouper species, as well as a new way of DNA identification to differentiate closely related species in fishes.     

一例引起青石斑鱼严重死亡的病因探讨

在福建罗源一例寄生虫,细胞,病毒综合感染引起的青石斑鱼Epinephelus awoara严重死亡的病例中,发现一种寄生于病鱼脾脏的病毒可能是引起死亡的主要致病因子,经电镜观察,这种病毒粒子为多面球状体,其平面观为六角形,大小约200nm,类似于真鲷虹彩病毒（Iridovirus)和鳜病毒（Siniperca chuastsi virus),青石斑鱼感染该病毒而引起严重死亡的病例的目前尚未见报导。     

斜带石斑鱼、赤点石斑鱼RAPD和线粒体Cyt b基因序列变异分析

斜带石斑鱼和赤点石斑鱼为具有重要经济价值的海水鱼类。对这两种石斑鱼进行了随机扩增多态DNA(RAPD)和线粒体细胞色素b(Cyt b)基因序列变异分析。计算多态位点百分率、基因多样性和香农信息指数等遗传参数,以此评估种内遗传变异水平;通过统计变异位点、平均核苷酸差异数、核苷酸多样性以及种间平均每位点核苷酸替代数进行基因序列变异分析,并构建UPGMA系统树。结果表明:斜带石斑鱼的遗传多样性水平高于赤点石斑鱼,这可能与它们在种内特定遗传结构、分布范围大小、自然资源状况的差异有关;斜带石斑鱼和赤点石斑鱼之间检测到的种特异RAPD条带以及基因序列的变异,可作为种间分子鉴定标记。     

海水鱼类寄生本尼登虫的ITS1 序列

从不同的海水鱼上采获一批本尼登虫(单殖纲、分室科、本尼登亚科)，其中部分标本经形态学研究后仍不能定种．本文采用PCR扩增及DNA序列测定的分子生物学方法，对棘鳞鳗本尼登虫、玫氏新本尼登虫、新本尼登虫未定种1和新本尼登虫未定种2的rDNA基因的TIS1序列进行了研究．结果表明：新本尼登虫未定种1和新本尼登虫未定种2与玫氏新本尼登虫的TIS1序列非常相似，差异率为0．7％(小于1％)，应为同一个种．而本尼登虫与新本尼登虫间的差异在33％以上．本研穷弥补了本尼得类单殖吸虫形态分类的某些不足．     

几种海水养殖鱼类化学组成的比较

对广东地区主要海水养殖鱼类青石斑鱼Epinephelus awoara、黄鳍鲷Sparus latus、银鲳Pampus argenteus、美国红鱼Sciaeunops ocellatus的全鱼及肌肉化学组成进行了测定.比较了几种鱼的肌肉水分、蛋白质、脂肪和灰分含量的差异,同时进一步检测了这几种鱼肌肉结构氨基酸的组成,为今后的营养研究提供必要的数据.     

RAPD Analysis for Genetic Variation within the Endangered Quillwort Isoetes hypsophila (Isoetaceae)

0　IntroductionI soetes hypsophila, a Chinese endemic species, is a memberof the heterosporous aquatic quillworts, and is a perennialdistributed in shallow zones of plateau lake or plateau marsh ofSichuan and Yunnan Province in China. In recent years, I.hypsophilahas declined rapidly in population quantity (num ber) and has even disappeared from many locations. The spe cies is now considered to be rare and threatened or endangeredin China and is listed among first category of protected wil…     

自然产卵的赤点石斑鱼胚胎及仔鱼形态发育研究

介绍赤点石斑鱼（Epinephelus akaara)人工种苗繁殖生产中,以自然产出的受精卵为材料,进行孵化、培育,连续观察该种鱼的胚胎发育过程及仔鱼的形态,并对其作的详细描述。     

南海北部近海5种石斑鱼的骨骼系统比较研究

为揭示骨骼系统在石斑鱼类的物种鉴定和系统进化关系研究中的应用潜力,本研究采用传统的骨骼系统比较法分析南海北部近海的尾纹九棘鲈(Cephalopholis urodeta)、鸢鮨(Triso dermopterus)、青石斑鱼(Epinephelus awoara)、拟青石斑鱼(E.fasciatomaculosus)和断带石斑鱼(E.craigi)等3属5种石斑鱼类的骨骼特征。结果显示,在分析的15种骨片中,中筛骨、额骨、翼耳骨、上枕骨、前鳃盖骨、上匙骨、尾舌骨等7种骨片显示出明显的属特征。尾纹九棘鲈的骨片形态结构总体较简单,而鸢鮨和石斑鱼属(Epinephelus)石斑鱼的骨片形态结构更为分化且复杂,并且二者具有相似类型的骨片数最多(8种)。有10种骨片可将断带石斑鱼与青石斑鱼、拟青石斑鱼有效区分,其中翼蝶骨、蝶耳骨、翼耳骨、上枕骨和前鳃盖骨等5种骨片在断带石斑鱼中显示出显著的种类特性。青石斑鱼和拟青石斑鱼的主要骨骼差异在于额骨、前鳃盖骨、上匙骨。本研究的结果支持了鸢鮨属(Triso)与石斑鱼属有较为密切的亲缘关系以及九棘鲈属(Cephalopholis)是较石斑鱼属更为原始的一个独...     

深圳梅林仙湖苏铁野生种群遗传多样性ISSR分析

仙湖苏铁Cycas fairylakeaD.Y.W ang是国家濒危保护植物,目前仅在我国广东省深圳市和曲江县发现有自然分布,个体总数已不足2 000株。本文应用ISSR标记对深圳梅林郊野公园仙湖苏铁野生种群的遗传多样性进行了研究。从100个引物中筛选出10个用于正式扩增,在14个个体中共检测到77个清晰的扩增位点,其中多态性位点67个,多态位点百分率为87.01%。根据所得数据求出的观察等位基因数为1.870 1,有效等位基因数为1.356 8,Ne i基因多样性指数为0.219 6,Shannon多样性指数为0.344 5。与其他苏铁类植物和濒危植物相比,仙湖苏铁仍然保持很高的遗传多样性水平。基于Jaccard相似性系数的UPGMA树系图表明,梅林郊野公园的仙湖苏铁个体明显地分为2个亚群,ISSR表型特征的主成分分析(PCA)也支持了聚类分析结果。研究结果对仙湖苏铁的保育和管理具有重要意义。     

蓝鳍金枪鱼和黄鳍金枪鱼遗传多样性的AFLP分析

利用选择性扩增片断长度多态性分子标记技术(Amplified fragment length polymorphism,AFLP),分析了采自日本九州海域蓝鳍金枪鱼(Thunnus thynnus)和台湾海域黄鳍金枪鱼(Thunnus albacares)野生群体的遗传多样性水平.结果表明:9 对选择性引物在 2 种金枪鱼中共扩增出675个位点(100～750 bp),其中蓝鳍金枪鱼和黄鳍金枪鱼多态位点分别为388个和368个.蓝鳍金枪鱼和黄鳍金枪鱼的多态位点比例、Shannon 遗传多样性指数分别为57.48%和54.52%,0.330 1和0.301 8.AMOVA分子方差分析显示:种间遗传分化中,83.79%的遗传变异由种间贡献,而16.21%的变异分布于种内个体之间.同其他鱼类比较,2 种金枪鱼显示了较为丰富的遗传多样性水平,其种质资源处于较好的水平.研究结果将为我国蓝鳍金枪鱼和黄鳍金枪鱼的资源保护与合理利用提供重要的理论依据.     

MHC II DRB variation and trans-species polymorphism in the golden snub-nosed monkey (Rhinopithecus roxellana)

Genetic variation is generally believed to be important in studying endangered species’ adaptive potential.Early studies assessed genetic diversity using nearly neutral markers,such as microsatellite loci and mitochondrial DNA(mtDNA),which are very informative for phylogenetic and phylogeographic reconstructions.However,the variation at these loci cannot provide direct information on selective processes involving the interaction of individuals with their environment,or on the capability to resist continuously evolving pathogens and parasites.The importance of genetic diversity at informative adaptive markers,such as major histocompatibility complex(MHC) genes,is increasingly being realized,especially in endangered,isolated species.Small population size and isolation make the golden snub-nosed monkey(Rhinopithecus roxellana) particularly susceptible to genetic variation losses through inbreeding and restricted gene flow.In this study,we compared the genetic variation and population structure of microsatellites,mtDNA,and the most relevant adaptive region of the MHC II-DRB genes in the golden snub-nosed monkey.We examined three Chinese R.roxellana populations and found the same variation patterns in all gene regions,with the population from Shennongjia population,Hubei Province,showing the lowest polymorphism among three populations.Genetic drift that outweighed balancing selection and the founder effect in these populations may explain the similar genetic variation pattern found in these neutral and adaptive genes.     

不同培水方式对斜带石斑鱼育苗池中浮游生物群落的影响

 斜带石斑鱼在中国的养殖主要位于南方,养殖规模仍不及其他海水养殖鱼类,如卵形鲳鲹、海鲈、军曹鱼等。苗种培育成活率低是造成斜带石斑鱼养殖业发展滞后的主要“瓶颈”之一,这与石斑鱼仔鱼开口期缺乏适宜的生物饵料有关,尤其是浮游动物的缺乏。为此,文中设计了4种不同培水方式,研究其对斜带石斑鱼育苗池中浮游生物群落的影响。方式1中分别添加益生菌、虾片40 mL·m^-3和4 g·m^-3（组1）,方式2中分别添加益生菌、虾片80 mL·m^-3和8 g·m^-3（组2）,方式3中添加小球藻液2.5 L·m^-3（组3）,方式4中添加小球藻液5 L·m^-3（组4）。每组设3个重复。育苗池规格2.5 m ×4.0 m×1.1 m（宽×长×高）。结果表明,组1和组2中浮游植物、浮游动物种类数高于组3和组4。益生菌和虾片的添加可显著提高育苗池中浮游植物及浮游动物密度。整个育苗过程中。组2所含浮游动物密度维持在较高水平(1.3 × 10.5~2.1 × 10.5 ind·L^-1),组1和组2中浮游动物密度要显著高于组3和组4(P≤0.05)。从实验中可看出石斑鱼育苗池中添加一定量的益生菌和虾片,可获得较丰富的浮游动物群落,为石斑鱼仔稚鱼生长提供活饵料。     

Development, chromosome location and genetic mapping of EST-SSR markers in wheat

A number of 151695 wheat expression sequence tags （ESTs） that originated from GenBank/dbEST from July 14, 2003 to August 24, 2004 were used to search for simple sequence repeats （SSRs） with motif 2-5 bp, and 2038 simple sequence repeats （EST-SSRs）, which accounted for 1.34% of EST database, were identified. Based on these SSR sequences, 249 EST-SSR primer pairs and 166 amplified clear bands in various wheat cultivars were designed. These EST-SSR markers can be used as new molecular markers in wheat and related species. Using Chinese Spring nulli-tetrasomic lines, 93 EST-SSR primer pairs and 193 EST-SSR loci were located on 19 wheat chromosomes except for 4A and 4B. Forty-three loci were mapped on 11 chromosomes of the genetic framework map previously constructed using recombinant inbred lines.     

赤点石斑鱼群体遗传结构的微卫星分析

 赤点石斑鱼Epinephelus akaara作为我国南方海域的经济水产鱼类,已面临着种质资源匮乏,过度开发的局面。为了开展其保护生物学研究和促进养殖业的健康发展,开展了赤点石斑鱼群体的微卫星分析,对中国东南沿海7个赤点石斑鱼群体的遗传变异进行了研究,发现在不同渔场之间的种群存在着丰富的遗传多样性,并且具有明显的地理区域分布特征,这为赤点石斑鱼的种质资源研究和遗传育种提供了遗传学基础。     

Mining,characterization, and exploitation of EST-derived microsatellites in <Emphasis Type="Italic">Gossypium barbadense</Emphasis>

Simple sequence repeats (SSRs) have been widely applied as molecular markers in genetic studies. However, the number of ex-pressed sequence tags (ESTs) and SSR markers from Gossypium barbadense is fewer than those from other cotton species. In this study, EST-SSR distribution from G. barbadense was characterized and new G. barbadense-derived EST-SSR markers were de-termined on the basis of the ESTs obtained by randomly sequencing 2 cDNA libraries associated with fiber development in G. barbadense. By mining 9697 non-redundant ESTs, a total of 638 SSR loci derived from 595 ESTs were observed. In G. barba-dense, the frequency of ESTs containing SSRs was 6.13%, with an average of 1 SSR in every 10.4 kb of EST sequence. Further-more, trinucleotide was found to be the most abundant repeat type among 2–6-nucleotide repeat types. It accounted for 26.6% of the total, followed by the hexanucleotide (26.0%) and pentanucleotide repeats (25.9%). Among all the repeat motifs, (AAG)n accounted for the highest proportion. EST-SSR primer pairs were developed using the Primer3 program, and the redundant primers were removed using the virtual PCR approach. As a result, 380 non-redundant EST-SSR primer pairs were developed and used to detect polymorphisms between the mapping parents G. hirsutum ‘TM-1’ and G. barbadense ‘Hai7124’ for constructing linkage groups in cultivated allotetraploid cotton. Out of these, 98 (25.8%) primer pairs detected polymorphisms. Finally, 95 polymorphic loci from 82 primer pairs were integrated into the backbone genetic map; of these, 42 were mapped into the A subgenome and 53 into the D subgenome. The present work provided the foundation for constructing saturated genetic maps and conducting comparative genomic studies on different cotton species.     

藏羚羊5个微卫星标记的多态性分析

藏羚羊是我国青藏高原的特有物种.为分析藏羚羊独特的遗传变异特征和核DNA遗传多样性,以5个微卫星标记对50只藏羚羊的等位基因多态性进行了研究.研究结果表明:5个微卫星标记共检测到92个等位基因,每个标记的平均等位基因数为18.4个(在16~23之间),平均有效等位基因数为11.1;5个微卫星标记的多态信息含量均在0.8573以上,为高度多态标记,其中L03标记的多态信息含量最高,达0.9372;各标记的观测杂合度在0.4898~0.9091之间,期望杂合度在0.8770~0.9504之间,平均期望杂合度为0.8990,属于高度杂合标记,遗传变异丰富.这些筛选出的多态性微卫星标记可应用于藏羚羊遗传多样性、遗传结构分析及遗传图谱的构建等工作.     

Development of simple sequence repeats (SSR) markers of ramie and comparison of SSR and inter-SSR marker systems

Ramie (Boehmeria nivea L. ) is an important bast fiber crop. To study genetic background of this species, we isolated and characterized microsatellite markers of ramie. A genomic library containing inserts of rapid amplification of polymorphic DNA (RAPD)fragments was constructed, and screened by PCR amplification using anchored simple sequence repeats as primers. A total of 26 clones were identified as positives, and 13 microsatellite loci were found after sequencing. The polymorphism of these 13 microsatellite loci was examined and the utility of simple sequence repeats (SSR) and inter-SSR (ISSR) marker systems for genetic characterization compared using 19 selected ramie cultivars. Both approaches successfully discriminated the 19 cultivars which differed in the amount of polymorphism detected. The level of polymorphism detected by SSR was 95.0 %, higher than that by ISSR (72.3 % ), but the average polymorphism information content (PIC) of ISSR (0. 651) was higher than that of SSR (0. 441). The higher PIC value of ISSR suggests that ISSR is more efficient for fingerprinting ramie cultivars than SSR markers. However, because the SSR loci are codominant, they are more suitable for determining the homozygosity levels of ramie, constructing linkage map, quantitative trait loci study of complex traits and marker-as-sisted selection.