Identification of a putative tetrasporophyte-specific gene in <Emphasis Type="Italic">Gracilaria lemaneiformis</Emphasis> (Gracilariales,Rhodophyte)

A putative tetrasporophyte-specific gene, designated as SSH466 (GenBank accession No. DQ019223), was one of the genes identified in this work using suppression subtractive hybridization (SSH) method in Gracilaria lemaneiformis. The full length of the gene was obtained using SMART RACE strategy. Sequence analysis revealed that the gene had 1 019 nucleotides, including an open reading frame of 498 nucleotides encoding 166 amino acid residues, 158 nucleotides of 5′ untranslated region and 363 nucleotides of 3′ non-coding region. Protein motif and secondary structure prediction showed that there existed a transmembrane domain with a unique β-sheet. Thus, SSH466 protein might be a cross-membrane protein. Sequence homology search in the public GenBank databases did not reveal any significant match with SSH466. Virtual Northern blot analysis confirmed that it was a tetrasporophyte-specific gene.     

Comparison in copper accumulation and physiological responses of Gracilaria lemaneiformis and G. lichenoides (Rhodophyceae)

Heavy metal pollution has become a worldwide problem in aquaculture. We studied copper (Cu2+ ) accumulation and physiological responses of two red algae Gracilaria lemaneiformis and Gracilaria lichenoides from China under Cu2+ exposure of 0-500 μg/L in concentration. Compared with G. lemaneiformis, G. lichenoides was more capable in accumulating Cu2+ , specifically, more Cu2+ on extracellular side (cell wall) than on intracellular side (cytoplasm) and in cell organelles (especially chloroplast, cell nucleus, mitochondria, and ribosome). In addition, G. lichenoides contained more insoluble polysaccharide in cell wall, which might promote the extracellular Cu2+ -binding as an efficient barrier against metal toxicity. Conversely, G. lemaneiformis was more vulnerable than G. lichenoides to Cu 2+ toxin for decreases in growth, pigment (chlorophyll a, chlorophyll b, phycobiliprotein, and β-carotene) content, and photosynthetic activity. Moreover, more serious oxidative damages in G. lemaneiformis than in G. lichenoides, in accumulation of reactive oxidative species and malondialdehyde, and in electrolyte leakage, because of lower antioxidant enzyme (superoxide dismutase and glutathione reductase) activities. Therefore, G. lichenoides was less susceptible to Cu2+ stress than G. lemaneiformis.     

Expression of the Phycoerythrin Gene of Gracilaria lemaneiformis (Rhodophyta) in E. coli and Evaluation of the Bioactivity of Recombinant PE

Phycoerythrin (PE) is one of the most important proteins involved in light capturing during photosynthesis in red algae. Its potential biological activities had gained wide concerns. In the present study, tumor cytotoxic and hydroxyl radical assay were preformed to detect the bioactivity of recombinant PE. Recombinant plasmids pGEX-PE and pBGL were transformed into E.coli BL21 to make two recombinant strains BEX (pGEX-PE) and BGL (pBGL). PE expressing in BEX (pGEX-PE) was validated by SDS-PAGE and Western blotting analysis. SDS-PAGE analysis indicated that the PE-GST fusion protein was mostly inclusion bod- ies. Specific expression of PE was confirmed by Western blotting analysis. The recombinant E. coli BEX (pGEX-PE) cells were col- lected and sonicated. The supernatants were reserved for the tumor cytotoxic experiments. The result of tumor cytotoxic assay indi- cated that the supernatants containing PE had the activity of inhibiting the growth of Hela cells and with the increase of protein con- centration, the inhibiting rate increased from 37.31% to 63.26%, which showed significant difference from the control. Hydroxyl radical scavenging effect was tested with supernatants of BEX (pGEX-PE) and BGL (pBGL) cell lysates treated with sonication and heating. For the sonication samples, the scavenging rates of the supernatants of BEX (pGEX-PE) and BGL (pBGL) cell lysates were significantly higher than the negative control BL21(pGEX-4T) (P<0.02), and the scavenging rates increased slowly following the increase of the protein content. For the heating samples, except for the 0.2 mg mL-1 BGL (pBGL) products, the scavenging effects of the supernatants of BEX (pGEX-PE) and BGL (pBGL) cell lysates were stronger than that of negative control BL21(pGEX-4T). However, the effect intensity was not positively correlated with the increase of the protein concentration. Though a partially de- creased hydroxyl radical scavenging activity was led by heating, the biological activity was still retained and conspicuous. This re- search showed that phycoerythrin protein expressing in E. coli has the potential medical and sanitarian value.     

Cloning, Expression and Characterization of Translationally Controlled Tumor Protein （TCTP） Gene from Flatfish Turbot （Scophthalmus maximus）

A full-length cDNA encoding translationally controlled tumor protein of marine flatfish turbot (Scophthalmus maximus), SmTCTP, was isolated with rapid amplification of cDNA Ends (RACE). SmTCTP consisted of a 5' untranslated region (UTR) of 84 bp, a 3' UTR of 451 bp and an open reading flame (ORF) of 513 bp, encoding a protein of 170 amino acid residues, which contained two signature sequences of TCTP family. The 5'UTR of SmTCTP started with a 5'-terminal oligopyrimidine tract (5'-TOP), a typical feature for translationaily controlled mRNAs. The deduced amino acid sequence of SmTCTP was similar to the other known verte-brate TCTPs in a range of 58.8% to 64.1%. The length offish TCTPs was diverse among species, e.g., TCTP of turbot and sea perch (Lateolabrax japonicus) is 170 aa in length, while that of zebrafish (Danio rer/o) and rohu (Labeo rohita) is 171 aa in length. North-ern blot analysis revealed that SmTCTP has only one type of mRNA. Its expression level in albino skin was slightly higher than that in normal skin. We constructed the pET3Oa-SmTCTP expression plasmid. The recombinant protein of His-tag SmTCTP was over-expressed in E. coli, purified and identified with peptide mass fingerprinting. These results may pave the way of further inves-tigation of the biological function of TCTP in fish.     

Identification and characterization of a new IgE-binding protein in mackerel (Scomber japonicus) by MALDI-TOF-MS

As fish is one source of the ‘big eight’ food allergens,the prevalence of fish allergy has increased over the past few years.In order to better understand fish allergy,it is necessary to identify fish allergens.Based on the sera from fish-allergenic patients,a 28 kDa protein from local mackerel (Scomber japonicus),which has not been reported as a fish allergen,was found to be reactive with most of the patients’ sera.The 28 kDa protein was analyzed by MALDI-TOF-MS (Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry).Mascot search in NCBI database (Date: 08/07/2010) showed that the top protein matched,i.e.triosephosphate isomerase (TPI) from Xiphophorus maculatus and Poecilia reticulata,had a mowse (molecular weight search) score of 98.In addition,TPI from Epinephelus coioides also matched this mackerel protein with a mowse score of 96.Because TPI is con-sidered as an allergen in other non-fish organisms,such as lychee,wheat,latex,archaeopotamobius (Archaeopotamobius sibiriensis) and crangon (Crangon crangon),we consider that it may also be an allergen in mackerel.     

注射黄芪多糖对吉富罗非鱼c型溶菌酶基因表达量的影响

将黄芪多糖(APS)用无菌生理盐水配制成2 mg/mL和20 mg/mL针剂,腹腔注射吉富罗非鱼,以注射无菌生理盐水为对照。24 h后分别提取吉富罗非鱼鳃、头肾、肝脏、脾脏等组织中的总RNA并反转录成cDNA,利用Real-time PCR方法对不同组织中基因表达进行定量分析。结果表明:吉富罗非鱼腹腔注射20 mg/mL高剂量APS后,其鳃、头肾、肝脏等三个组织中的Lysozyme-c基因表达量显著高于对照组(P<0.05);注射2 mg/mL低剂量APS后,Lysozyme-c基因表达量仅在脾脏中出现显著上调(P<0.05)。APS可通过诱导Lysozyme-c基因在鳃、头肾、肝脏和脾脏等组织在的表达量,来提高吉富罗非鱼的机体免疫力。     

Biomass and carbon storage of Gracilariopsis lemaneiformis （Rhodophyta） in Zhanshan Bay,Qingdao, China

Marine macroalgae can absorb carbon and play an important role in carbon sequestration. As an important economic macroalga, Gracilariopsis lemaneiformis has the potential to significantly affect carbon absorption and storage in wave-sheltered intertidal reef systems. However, detailed knowledge on seasonal biomass changes and carbon storage of G. lemaneiformis is lacking, especially in many small and scattered ecosystems. Considering the influence of human activities on wild distribution of G. lemaneiformis, the understanding of seasonal dynamics of an economically important species in nature is necessary. In this study, we first investigated seasonal variations in biomass, coverage area, and carbon storage during low tide from August 2011 to July 2012 in Zhanshan Bay, Qingdao, China. Furthermore, we estimated the carbon storage potential of wild G. lemaneiformis using light use efficiency(LUE). The results show that the standing biomass and coverage area changed significantly with season. However, seasonal variations in carbon content and water content were not obvious, with an average content of 35.1% and 83.64%, respectively. Moreover, carbon storage in individual months varied between 0.67 and 47.03 g C/m 2, and the value of carbon storage was the highest in August and June and the lowest in February. In Zhanshan Bay, LUE of G. lemaneiformis was only 0.23%. If it is increased to the theoretical maximum(5%–6%), the carbon storage will have an increase of at least 21 times compared with the current, which suggested that carbon storage of wild G. lemaneiformis had a high enhancement potential. The study will help to assess a potential role of G. lemaneiformis in reducing atmospheric CO2.     

红笛鲷重组激活基因rag1原核表达条件的优化及纯化

克隆编码红笛鲷(Lutjanus sanguineus)RAG1蛋白(recombination activating protein1)活性核心区的基因序列,并与pET-28a(+)载体连接,构建原核表达载体pET-28a-RAG1,将其转入大肠杆菌BL21(DE3)菌株,利用IPTG进行诱导表达。为提高融合蛋白的表达效率,运用传统的实验方法对诱导条件进行优化。SDS-PAGE分析表明,在37℃条件下,利用0.1 mmol/L IPTG诱导8 h后,RAG1重组融合蛋白的表达量最大,相对分子质量与预测值相符,该蛋白主要以包涵体形式高效表达,利用His Trap HP亲和柱使其得到进一步纯化;Western blot分析显示,该融合蛋白可与鼠抗His-tag单克隆抗体发生特异性结合,说明表达蛋白为目的蛋白。     

Polychlorinated dibenzo-p-dioxins （PCDDs） and dibenzofurans （PCDFs） in surface sediment and bivalve from the Changjiang Estuary, China

Surface sediments and bivalves were collected from the Changjiang Estuary in December 2003 and November 2004, respectively. Polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs) in these samples were measured with high-resolution chromatography (HRGC)/High Resolution Mass Spectrometer (HRMS). The concentrations of total PCDD/Fs and toxic equivalent (TEQ) were 169.83±119.63 and 0.81±0.36 pg/g dry weight (dw) in sediments, and 580.33±240.17 and 7.24±3.65 pg/g dw in bivalves. The homolog compositions of PCDD/Fs were similar among samples, the most abundant congener was octa-chlorinated dibenzo-p-dioxin (OCDD) and then octa-chlorinated dibenzofuran (OCDF) and 1,2,3,4,6,7,8-hepta-chlorinated dibenzo-p-dioxin (HpCDD). The herbicide pentachlorophenol (PCP) and sodium pentachlorophenol (Na-PCP) were proved the main source of PCDD/Fs in this area.     

Characteristics of carbonaceous aerosol in a two-week campaign at Ny-Alesund

The concentrations of organic carbon (OC) and elemental carbon (EC)in total suspended particle (TSP) were investigated at Ny-(A)lesund, Svalbard in a two-week campaign. The levels of OC and EC are 0.86 ±0.27 μgm-3 (mean±standard deviation) and 0. 19±0.10 μgm-3 , respectively. Back trajectory analysis of air masses arriving at Ny-(A)lesund reveals that long-range transport of polluted air play insignificant role in OC and EC levels, to which the potential influence of the local contamination were ascribed. The average OC/EC ratio is 5.41,suggesting the presence of the secondary organic aerosols. The estimated secondary organic carbon (SOC) in TSP is 0.59 μg/m3 , accounting for 64% of the total organic carbon.     

Investigation of a new vortex-induced vibration suppression device in laboratory experiments

In order to mitigate vortex-induced vibration (VIV) of marine risers, especially to eliminate the phenomenon of frequency ’lock-in’, a new suppression device of crescent-shaped flow spoiler was designed with seven different layout schemes. VIV model tests with six flow levels were conducted in a large wind-wave-current flume. In all cases, vibration responses in both in-line and cross-flow cases were measured. With the installation of suppression devices vibration frequency evolution of a riser was analyzed by Morlet wavelet transform. The principle of VIV suppression was interpreted through vibration characteristics. Fatigue life of the riser was calculated by the Palmgren-Miner rule. Compared with a bare riser, vibration of an outfitted riser with suppression devices disturbed the steady flow, the vibration amplitudes in the two flow directions were reduced, and the riser fatigue life was improved.     

企鹅珍珠贝生殖细胞的发生

对不同发育阶段的企鹅珍珠贝[Ptedapenguin(Rding)]的性腺结构、生殖细胞结构与分布进行组织学观察,描述精/卵子发生过程中各级精/卵细胞的特征。结果表明:根据雌性生殖细胞形态和卵黄的变化特点,卵子发生经历卵原细胞、无卵黄初级卵母细胞、卵黄形成期卵母细胞及成熟卵子阶段;依据雄性生殖细胞大小和细胞核形态变化,精子发生经历精原细胞、初级精母细胞、次级精母细胞、精子细胞和精子阶段,其中精原细胞存在形态上有差异的A型和B型两种类型。此外还观察到企鹅珍珠贝的性别转化现象。     

Optimization of fluorescence in situ hybridization （FISH） for the identification of two polar coccoid green algae species

Standard FISH protocols using fluorochrome-labeled oligonucteotide probes have been successfully applied for in situ detection.However,optimized protocols of FISH for specific eukaryotes in marine environments are often not developed.This study optimized the conditions of fluorescence in situ hybridization (FISH) by using two polar isolated microalgae.The modified conditions were as follows: (1) 10 mg·mL-1 lysozyme solution pretreatment at 37℃ for 30 min;(2) the hybridization buffer including 20% formamide;(3) the hybridization condition was 47℃ for 6 h. The cells enumerated by FISH were compared with those enumerated by flow eytometry (FCM) and DAPI to confirm the cell loss and hybridization efficiency.The optimized protocol was also successfully applied to Arctic Ocean samples,which were found to be dominated by Micromonas sp.The modified protocol showed a high relative efficiency and could be successfully applied for the detection of specific microbial eukaryotes in environmental samples.     

荷那龙罗非鱼两种GHSR基因的克隆与序列分析

用RT-PCR和RACE方法,从荷那龙罗非鱼(Oreochromis hornorum)垂体中克隆到生长激素促分泌素受体(GHSR)cDNA全序列。荷那龙罗非鱼GHSR基因具有GHSR-1a与GHSR-1b两个高度保守cDNA序列。GHSR-1a序列全长1 646 bp,包括225 bp的5′非编码区,266 bp的3′非编码区和1 155 bp的开放阅读框,编码384个氨基酸残基,具有7个跨膜结构域结构(transmembrane domains,TM);GHSR-1b序列全长1 877 bp,包括225 bp的5′非编码区,755 bp的3′非编码区和897 bp的开放阅读框,编码298个氨基酸残基,只具有前5个TM,在第6个TM的第4个氨基酸处开始缺失。将GHSR cDNA序列与基因组序列比较发现,这两种cDNA转录本来自同一个基因的不同变体。     

Comparative fatty acid composition of fourSargassum species (Fucales,Phaeophyta)

Fatty acid composition of fourSargassum species from Qingdao and Shidao, Shandong Province was investigated. 16:0 (palmitic acid) was the major saturated fatty acid. C₁₈ and C₂₀ were the main polyunsaturated fatty acids (PUFAs). Arachidonic acid and eicosapentaenoic acid predominated among polyenoic acids in all the algal species examined, except forSargassum sp. which had low concentration of eicosapentaenoic acid. Contribution No. 2354 from the Institute of Oceanology, Chinese Academy of Sciences     

PSO-GM(1,1,N,p,ξ)模型在变形预测中的应用

通过对传统GM(1,1)缺陷分析和改进的基于权的PGM(1,1)建模机理描述,顾及PGM(1,1)中背景值构造时取相同的参数不能充分降低模型的预测误差,对不同的时刻引入不同的参数来改进GM(1,1)背景值序列的计算公式,将这种背景值构造方法和灰元N引入GM(1,1)建立了新的白化方程。在建立的新的白化方程基础上,用龙格-库塔法以修正的初始值计算累加值的模拟序列。针对引入的参数较多问题,采用粒子群算法寻找满足相对误差均值最优的参数,从而建立了基于粒子群优化算法和加权灰色组合的PSO-GM模型。工程实例应用表明,新模型的拟合精度高,预测效果好,相对其他两种原有模型预测精度有明显提高。     

Removal of eutrophication factors and heavy metal from a closed cultivation system using the macroalgae, Gracilaria sp. (Rhodophyta)

In this study,the ability of macroalgae Gracilaria sp.of removing eutrophication factors and toxic heavy metals Al,Cr,and Zn in a closed cultivation system is reported.The results show that the concentration of the three heavy metals decreased significantly during the experimental period in an algal biomass dependent manner.The biofiltration capacity of the alga for Al,Cr,and Zn is 10.1%-72.6%,52.5%-83.4% and 36.5%-91.7%,respectively.Using more materials resulted in stronger heavy metal removal.Additionally...     

IDENTIFICATION OF CYTOKININ IN A GREEN ALGAE EXTRACT

Isopentnyladenosine(i~6Ado) was identified, and trans-zeatin (trans-Z) and trans-zeatin riboside(trans-ZR) were detected by high pressure liquld chrmoatoguphy (HPLC) but not verified withchromatography - mass spectrometry (GC - M S ) analysis of cytokinin from the extracts of green algae (Ulvapertusa (Kjellm), Enteromopha compressa and Monostma sp). This indicated that the green algae mixturecontained cytokinin -like substances.     

First record of Paraonella platybranchia （Hartman, 1961） （Polychaeta： Paraonidae） from the Yellow Sea

This paper reports for the first time a species of polychaetous family Paraonidae, Paraonella platybranchia (Hartman, 1961), a rare species found previously only in the coast near the boundary between the United States and Mexico, and it was recently discovered in the samples collected in 1982 from Chinese side of the Yellow Sea. This is the first record of the species in China as well as in the Northwest Pacific.