养殖大黄鱼(Pseudosciaena crocea)3种致病弧菌的分子鉴定及其系统发育学分析

从患病养殖大黄鱼分离到3株病原菌H040823-1、H050704-1、H050815-1,经常规生理生化鉴定均属于弧菌属的种类,API20E快速鉴定菌株H040823-1为副溶血弧菌(Vibrio parahaemolyti-cus),菌株H050815-1为溶藻弧菌(V.alginolyticus),菌株H050704-1不在API20E鉴定谱内。为了进一步确定其分类地位,测定了3株病原菌的16S rRNA和HSP60(heat shock protein,HSP60)基因部分序列。16S RNA基因系统进化分析表明,3株病原菌与副溶血弧菌、哈维氏弧菌、溶藻弧菌亲缘关系较近,相互之间同源性均大于96.9%,差异不明显。HSP60基因序列分析表明,菌株H040823-1、H050704-1、H050815-1的HSP60基因序列分别与V.parahaemolyticus(AF230951)、V.harveyi(EU036994)、V.alginolyticus(DQ664545)的同源性最高,分别为96.7%、99.8%和98.0%,而与其他弧菌HSP60基因的同源性均低于91.9%,3株病原菌相互之间同源性低于92.3%,差异显著。HSP60基因构建的系统进化树表明,H040823-1、H050704-1、H050815-1分别与V.parahaemolyticus、V.harveyi、V.alginolyticus聚类。综合以上结果,菌株H040823-1、H050704-1、H050815-1可分别鉴定为副溶血弧菌、哈维氏弧菌、溶藻弧菌。结果表明,HSP60基因比16S rRNA基因更适合用于海水鱼类致病性弧菌种间的分类研究。     

诱导子对海洋真菌Penicillium crustosum 2A4-Z15菌株生物碱代谢的调节作用

分别采用4种生物源诱导子和5种非生物源诱导子,对具有较强抑菌活性的海洋真菌壳青霉Penicillium crustosum2A4-Z15菌株进行诱导,考察诱导子对于菌株产生物碱的代谢调节作用。结果表明,不同来源的诱导子对菌株的生物碱产量和生物量影响不同,生物源诱导子的正向调节作用较明显,且其中以细菌源诱导子最为明显。当海洋细菌源诱导子在2A4-Z15菌株发酵对数生长期末,以10μg/cm3的浓度加入时,发酵物生物碱产量最高,较对照提高44.1%。     

白斑综合症病毒膜蛋白VP124参与病毒的感染

白斑综合症病毒(white spot syndrome virus,WSSV)是对虾的最主要病毒病原之一,给对虾养殖造成了巨大的经济损失。WSSV分别与三种该病毒膜蛋白VP39,VP124和VP187的特异性抗血清在体外作用后,再注射到螯虾体内,进行中和试验。结果显示,注射同或未同VP124抗血清作用的WSSV,8 d后螯虾的死亡率分别是100%和60%,WSSV的感染能够被抗VP124抗体中和。进一步利用定量PCR分析上述三种特异性抗血清对病毒感染的影响,结果显示,VP124抗血清可抑制WSSV在螯虾体内的增殖。所有结果表明,VP124在病毒的感染中起作用。     

Underwater measurements of carbon dioxide evolution in marine plant communities: A new method

We developed a new methodology to determine CO₂ fluxes in intertidal and shallow subtidal plant communities, namely seagrasses, both when the plants are submerged and when they are air-exposed. The apparatus comprises closed incubation chambers and a gas exchange column, designed to remove carbon dioxide from the water. Different types of incubation chambers were designed and built to adapt the system to distinct environments and incubation requirements. The methodology was tested under a comprehensive range of situations and its advantages and limitations are discussed. Overall, the method provides precise measurements of community carbon dioxide fluxes, through a fast and non-intrusive process, allowing repeatable in situ measurements of carbon uptake both in submerged and air-exposed conditions. As the experimental apparatus is identical, directly comparable measurements of air-exposed and submerged community production may be obtained, allowing sound estimates of daily carbon budgets of intertidal and shallow subtidal communities.     

Organic matter dynamics and budgets in the turbidity maximum zone of the Seine Estuary (France)

Organic matter was studied in the turbidity maximum zone (TMZ) of the Seine Estuary during 8 tidal cycles from April to October in 2001, 2002 and 2003, covering a salinity range from 0 to 27. The hydrological conditions were quite varied (extremely wet in 2001, unusually dry in 2003). A particularly striking feature is the high organic matter content in the suspended solids (SS) of the Seine estuary (4–5%).     

Modelling ecosystem functions and properties at different time and spatial scales in shallow coastal lagoons: An application of the LOICZ biogeochemical model

The Land-Ocean Interactions in the Coastal Zone (LOICZ) biogeochemical model (LBM) was applied at different temporal and spatial scales in 17 Italian lagoons of the LaguNet network (http://www.dsa.unipr.it/lagunet). A series of alternative assumptions taking into account benthic vegetation and sedimentary fluxes were introduced and compared with the classical LBM approach at various time scales. The reliability of the LBM application to the seventeen Italian lagoons was tested by comparison to a pool of shallow coastal systems from the global LOICZ database with comparable depths and sizes. The nutrient loads of the Italian sites can be considered relatively low, particularly for dissolved inorganic phosphorus (DIP). Although the median values of estimated internal transformations (source-sink) of both dissolved inorganic phosphorous and nitrogen at the LaguNet sites were comparable with the selected LOICZ sites, the positive and negative extreme values were one order of magnitude lower. Overall, the LBM applications to the Italian sites gave good quality budgets for shallow systems subjected to relatively low nutrient inputs and with a wide range of primary producer communities, including seagrass, macroalgae and phytoplankton. Furthermore, stoichiometry of Carbon:Nitrogen:Phosphorous for the different primary producer groups allowed the integration of previous studies by identifying a series of relationships between nutrient loads and ecosystem functions.     

S7核糖体蛋白基因片段序列变异与花鲈属鱼类系统发育研究

对中国和日本海域花鲈属的3个种:花鲈(Lateolabrax maculatus)、鲈鱼(L. japoni-cus)和高体鲈(L. latus)的S7核糖体蛋白基因片段序列进行了扩增和序列测定,分析比较了3个种23个个体间的序列差异。在456bp的S7核糖体蛋白基因片段中,3个高体鲈个体中出现了3种单倍型,种内个体间有2个碱基差异;7个花鲈个体中出现了7种单倍型,种内个体间有21个碱基差异;13个鲈鱼个体中出现了12种单倍型,种内个体间有22个碱基差异。结果表明,花鲈属S7核糖体蛋白基因片段序列具有丰富的遗传信息,在亲缘关系较近的物种间也存在显著的序列差异,基于S7核糖体蛋白基因片段序列的NJ和ME系统树经1000次重复抽样检验后得到相似结果,表明S7基因内含子2是适合于研究分子系统发育的分子标记。利用Modeltest选取最佳核苷酸替代模型构建的NJ树表明:花鲈属鱼类由高体鲈分化,花鲈与鲈鱼的亲缘关系很近,而与高体鲈的亲缘关系较远。     

Cloning and sequence analysis of a full-length cDNA of SmPP1cb encoding turbot protein phosphatase 1 beta catalytic subunit

Reversible protein phosphorylation, catalyzed by protein kinases and phosphatases, is an important and versatile mechanism by which eukaryotic cells regulate almost all the signaling processes. Protein phosphatase 1 (PP1) is the first and well-characterized member of the protein serine/threonine phosphatase family. In the present study, a full-length cDNA encoding the beta isoform of the catalytic subunit of protein phosphatase 1(PP1cb), was for the first time isolated and sequenced from the skin tissue of flatfish turbot Scophthalmus maximus, designated SmPP1cb, by the rapid amplification of cDNA ends (RACE) technique. The cDNA sequence of SmPP1cb we obtained contains a 984 bp open reading frame (ORF), flanked by a complete 39 bp 5' untranslated region and 462 bp 3' untranslated region. The ORF encodes a putative 327 amino acid protein, and the N-terminal section of this protein is highly acidic, Met-Ala-Glu-Gly-Glu-Leu-Asp-Val-Asp, a common feature for PP1 catalytic subunit but absent in protein phosphatase 2B (PP2B). And its calculated molecular mass is 37 193 Da and pI 5.8. Sequence analysis indicated that, SmPP1cb is extremely conserved in both amino acid and nucleotide acid levels compared with the PP1cb of other vertebrates and invertebrates, and its Kozak motif contained in the 5'UTR around ATG start codon is GXXAXXGXXATGG, which is different from mammalian in two positions A-6 and G-3, indicating the possibility of different initiation of translation in turbot, and also the 3'UTR of SmPP1cb is highly diverse in the sequence similarity and length compared with other animals, especially zebrafish. The cloning and sequencing of SmPP1cb gene lays a good foundation for the future work on the biological functions of PP1 in the flatfish turbot.     

Efficient expression of green fluorescent protein （GFP） mediated by a chimeric promoter in Chlamydomonas reinhardtii

To improve the expression efficiency of exogenous genes in Chlamydomonas reinhardtii,a high efficient expression vector was constructed.Green fluorescent protein(GFP) was expressed in C.reinhardtii under the control of promoters:RBCS2 and HSP70A-RBCS2.Efficiency of transformation and expression were compared between two transgenic algae:RBCS2 mediated strain Tran-Ⅰ and HSP70A-RBCS2 mediated strain Tran-Ⅱ.Results show that HSP70A-RBCS2 could improve greatly the transformation efficiency by approximately eightfold of RBCS2,and the expression efficiency of GFP in Tran-Ⅱ was at least double of that in Tran-Ⅰ.In addition,a threefold increase of GFP in Tran-Ⅱwas induced by heat shock at 40°C.All of the results demonstrated that HSP70A-RBCS2 was more efficient than RBCS2 in expressing exogenous gene in C.reinhardtii.     

恩诺沙星在凡纳滨对虾体内的代谢和残留消除规律

在26±2℃水温下,每天投喂含有恩诺沙星药物的饲料,研究恩诺沙星在凡纳滨对虾Litopenaeus vanname肌肉、肠和肝胰脏组织中的代谢和残留消除规律。残留药物用乙腈提取,液相色谱串联质谱仪检测。结果表明:恩诺沙星在凡纳滨对虾体内可代谢为环丙沙星,对虾体内同时有恩诺沙星和环丙沙星两种药物残留;环丙沙星在肌肉、肝胰脏和肠组织中的消除时间分别为6、8、10 d,而恩诺沙星在这三组织中的消除时间则为12、14、16 d。建议把肠作为该药残留监控的靶组织,凡纳滨对虾的休药期不少于16 d。