牙鲆CA/GT微卫星标记的筛选

采用生物素选择杂交法与放射性同位素杂交法相结合的技术,成功地从牙鲆(Paralichthys olivaceus)基因组中分离出含有CA/GT重复类型的微卫星序列。通过两轮淘选,共获得526个阳性菌落。测序其中的119个菌落,结果获得133个含有微卫星座位的序列。除了两个复合型微卫星外(1.5%),完美型63个(47.37%),非完美型68个(51.13%)。设计并合成22对微卫星引物,对8个人工雌核发育家系的亲本进行遗传背景分析。PCR结果表明,4对引物无扩增带或者扩增带不是目的条带,1对引物表现为单态,其余17对引物均呈多态性,平均每个座位产生5.2个复等位基因,杂合度为0.375～0.846,多态信息含量为0.305～0.823。结果表明,所筛选的大部分微卫星标记能够用于牙鲆群体遗传学研究。     

史氏鲟免疫球蛋白重链可变区序列及多样性

为了研究史氏鲟免疫球蛋白重链可变区基因的组织结构和多样性,采用RTPCR技术从史氏鲟(Acipenserschrenckii)脾脏总RNA中获得了免疫球蛋白重链可变区cDNA克隆,随机挑取31个阳性克隆进行测序。结果表明:所有序列相同率高于75%,前导肽相同率高于90%,应属于同1个VH家族。其变异主要存在于互补性决定区,特别是CDR3区。在D片段序列中发现大量保守的基因序列(motif)。并发现多个VH基因片段可以共用一个J片段的现象。在基因组DNA重排过程中,VH片段可以与任意的D和J片段结合。此外,史氏鲟免疫球蛋白重链可变区的VH,D和J片段的随机重排外,外切核酸酶作用,以及在重排位点大量N,P片段的插入现象,都大大增加了鲟鱼免疫球蛋白的多样性。     

Characterization of a novel nm23 gene and its potential roles in gametogenesis in the prawn Macrobrachium rosenbergii (de Man, 1879) (Crustacea: Decapoda)

Nm23 is a family of genes encoding the nucleoside diphosphate (NDP) kinase, which functions in a wide variety of biological processes, including growth, development, differentiation and tumor metastasis. In this study, a novel nm23 gene, designated as Mrnm23, was identified from the freshwater giant prawn Macrobrachium rosenbergii. The full-length cDNA was 776 bp in length, encoding for a protein of 176 amino acids with one typical NDP kinase domain that harbored all the crucial residues for nucleotide binding and enzymatic activity. Like human novel nm23-H1B, the putative protein contained a unique 21-amino-acid NH₂-terminal extension as compared to human nm23 (nm23-H1) homologs. Further, 3 extra amino acid residues prolonged the COOH-terminus. The Mrnm23 was ubiquitously expressed in all tissues examined, including androgenic gland, gill, heart, liver, muscle, ovary, and testis. In situ hybridization to gonad sections indicated that the Mrnm23 mRNA was localized in the cytoplasm of cup-base of differentiating spermatids, in the spike of the umbrella-shaped spermatozoa and in the cytoplasm of the early previtellogenic oocytes, suggesting that the Mrnm23 has potential roles in spermiogenesis and early differentiation of oocyte.     

Molecular cloning and sequence analysis of methionine adenosyltransferase from the economic seaweed Undaria pinnatifida

The methionine adenosyltransferase gene (MAT) had been isolated from an economic seaweed Undaria pinnatifida by PCR using degenerate primers. The cDNA was 1,491 bp in length with an open reading frame of 1,194 nucleotides, encoding a deduced protein of 397 amino acids. The protein had a predicted molecular weight of 43.2 kDa, and the isoelectric point was 5.244. The sequence contains a 92 bp 5′-untranslated region (UTR) and a 205 bp 3′-UTR. The methionine adenosyltransferase (MAT) sequence of U. pinnatifida (UpMAT) shared 68–92 % identities with the previous published MAT sequences of other species. Phylogenetic analysis indicated that the phylogenetic relationship of UpMAT with some other seaweeds was closer than with those of higher plants. Under different stress conditions, the relative mRNA expression levels of the MAT of U. pinnatifida (UpMAT) were measured by real-time quantitative PCR, and the results demonstrated that the UpMAT might help to protect the alga against various abiotic stresses.     

A study on early tolerance of Mactra chinensis philippi to salinity

In order to evaluate the early tolerance of Mactra chinensis to salinity, the treatments of salinity gradients and salinity gradual changes were set in this study, and the post growth and development of juveniles were analyzed in recovery experiment, respectively. The result showed that the optimum hatching of zygotes was found at a salinity from 24 to 32, which is narrower than that of larvae (20–32); a slight of low salinity (16–32) will benefit the early growth and development of M. chinensis; at planktonic and creeping stages, low salinity stress (20) was conducive to promoting the growth of juvenile M. chinensis philippi; 4, 48 was the ultimate salinity of M. chinensis; The range of early tolerance of larvae M. chinensis philippi to salinity can be widened through a short period of salinity acclimation.     

温度和流速对洛氏鱥呼吸代谢的影响

采用实验生态学的方法研究了不同温度和流速对两个年龄组洛氏鱥呼吸代谢的影响。结果表明,1龄和2龄洛氏鱥在4℃水体中基本不摄食生长十分缓慢,随着水体温度升高,鱼类摄食和生长情况出现好转。在温度4℃增高至28℃的过程中,除流速16cm/s和18cm/s条件下水温由24℃升28℃时耗氧率出现降低外,其余流速下洛氏鱥耗氧率均随着温度升高而增加,呈典型的指数增长趋势Y=aebx,R2值均高于0.90;在流速由8cm/s增至18cm/s的过程中,虽然1龄组在低于20℃和2龄组低于24℃水体中耗氧率随流速增加而增加,但1龄组在温度24、28℃和2龄组在28℃的水体中当流速增至16cm/s时耗氧率达到最大值,1龄组持续30min和2龄组持续1h后出现鱼体紧贴栏鱼网的现象。通过分析得出,洛氏鱥的呼吸代谢受到温度、水流和个体大小等因素的相互影响,个体较大的2龄洛氏鱥对水环境的适应范围明显强大于1龄鱼,流速为12—14cm/s温度为16—24℃和流速为16cm/s温度为16—20℃是1龄洛氏鱥理想养殖条件,流速12—16cm/s温度8—24℃是2龄洛氏鱥理想养殖条件。     

不同饵料对匙吻鲟仔鱼生长发育和消化酶活性的影响

本文研究了不同强化饵料对匙吻鲟(Polyodon spathala)仔鱼生长和发育的影响,实验采用蛋黄或鱼油强化卤虫无节幼体或桡足幼体为饵料的共6个实验组,即未强化卤虫组、蛋黄强化卤虫组、鱼油强化卤虫组和未强化桡足幼体组、蛋黄强化桡足幼体组、鱼油强化桡足幼体组,对匙吻鲟仔鱼开口驯化14 d,动态监测和统计不同饵料组匙吻鲟仔鱼的生长和存活情况,并对消化酶活性进行相关性分析。体重、体长、日增重和存活率4个指标,均为桡足幼体组显著高于卤虫组(P0.05),尤其在存活率方面,鱼油强化卤虫组不足50%,而未强化桡足幼体组最高可达86.59%;不同饵料组生长模式方程都获得较好的拟合,从体重和体长生长曲线看,桡足幼体组从饲喂8天起体长和体重进入快速生长期,而卤虫组生长一直相对缓慢;胃蛋白酶活性在未强化桡足幼体组显著高于其他组(P0.05),不同饵料组对仔鱼的淀粉酶活性无显著性影响。结果表明,在匙吻鲟仔鱼开口期以桡足幼体开口饵料驯化效果较好,特别是未强化桡足幼体组仔鱼的存活率高,鱼油强化桡足幼体组仔鱼生长速度较快,而以卤虫饲喂效果相对较差。     

NaCl盐度和NaHCO3碱度对松浦镜鲤、方正鲫的孵化及其仔鱼存活的影响

研究了NaCl盐度(0、1、3、5、7、9)和NaHCO3碱度(1.00 mmol/L、10.00 mmol/L、15.85 mmol/L、25.12 mmol/L、39.81 mmol/L、63.10 mmol/L)对松浦镜鲤(Cynipus carpio)与方正鲫(Carassius auratus gibelio)的孵化率、畸形率以及仔鱼24 h和48 h存活率的影响。1盐度实验结果,松浦镜鲤在盐度为0、1时,受精卵的孵化率较高,平均为72.40%±6.09%;在盐度0~3时,孵出仔鱼畸形率较低,平均为6.02%±1.87%,48 h存活率较高,平均为94.99%±6.44%。方正鲫在盐度0~3时,受精卵的孵化率较高,平均为68.03%±15.44%;在盐度0~5时,孵出仔鱼畸形率较低,平均为7.95%±3.24%,48 h存活率较高,平均为88.52%±9.19%。2碱度实验结果,这两种鱼的孵化率在碱度1.00~25.12 mmol/L时较高,松浦镜鲤平均为47.96%±8.43%,方正鲫平均为66.23%±18.11%。畸形率在碱度1.00~15.85 mmol/L范围内较低,松浦镜鲤平均为12.47%±3.77%,方正鲫平均为12.46%±8.44%。48 h存活率在碱度1.00~15.85 mmol/L时较高,松浦镜鲤平均为96.16%±5.04%,方正鲫平均为85.62%±3.73%。综合各项指标,松浦镜鲤和方正鲫人工孵化用水建议盐度控制在3以内,碱度控制在15 mmol/L以内。     

大黄鱼耐低温性状相关微卫星标记的筛选

鱼类的耐低温性状是一种重要的经济性状。为了初步分析大黄鱼的耐低温性状, 文章采用15对荧光微卫星标记, 以SSR-PCR方法对大黄鱼低温耐受组和正常对照组F₁代共40个个体进行了耐低温性状遗传差异分析。结果显示, 标记LYC0002在两组样品中共扩增出5个等位基因(片段大小分别为112 bp、110 bp、108 bp、106 bp和104 bp), 其中LYC00021_{12 bp}等位基因在低温耐受组的出现频率达60%, 而在正常对照组中的频率为零, 表明该等位基因对大黄鱼的温度敏感特性有较明显的偏好性, 可能与某种耐低温基因存在一定的连锁关系。此外, 对LYC0002_{106 bp、}LYC0002_{108 bp}、LYC0002_{110 bp} 和 LYC0002_{112 bp} 4个等位基因分别进行了回收、克隆及测序。序列比对结果显示, LYC0002_{112 bp}等位基因含有10个(CA)重复单元, 而其他3个等位基因依次缺失1个(CA)重复单元, 说明LYC0002在本研究样本中的突变方式为微卫星逐步突变模型(Stepwise mutation model, SMM)。     

大西洋鲑杀鲑气单胞菌无色亚种的分离鉴定和致病性研究

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