The cDNA cloning and mRNA expression of a potential selenium-binding protein gene in the scallop Chlamys farreri

Selenium binding proteins (SeBP) represent a family of proteins that are believed to be involved in controlling the oxidation/reduction in many physiological processes. The cDNA of Zhikong Scallop Chlamys farreri selenium binding protein (zSeBP) was cloned by expressed sequence tag (EST) and RACE techniques. The high similarity of zSeBP deduced amino acid sequence with the SeBP in other organisms, such as bird, fish, frog, mosquito, fruit fly, mammalian, and even nematode and microorganism indicated that zSeBP should be a member of SeBP family. The temporal expression of zSeBP in the hemocytes was measured by semi-quantitative RT-PCR after scallops were stimulated by either oxidative stress or microbial challenge. The expression of zSeBP was up-regulated progressively after stimulation, and then dropped gradually to the original level. Meanwhile, malondialdehyde (MDA) measured by the colorimetric method in the microbial challenged scallops increased immediately after scallops was challenged by microbes, and was significantly higher than that in the control scallops. Results indicated that the microbial infection could incense the disorder of oxidation/reduction and may result in high MDA production. The negative correlation between the expression level of zSeBP and the MDA content suggested that zSeBP could play an important role in mediating the anti-oxidation mechanisms and immune response in marine invertebrates. (c) 2005 Published by Elsevier Ltd.

The cDNA cloning and mRNA expression of heat shock protein 70 gene in the haemocytes of bay scallop (Argopecten irradians, Lamarck 1819) responding to bacteria challenge and naphthalin stress

Heat shock protein 70 (HSP70) is an important member of the heat shock protein superfamily, and it plays a key role in the process of protecting cells, facilitating the folding of nascent peptides and responding to stress. The cDNA of bay scallop Argopecten irradians HSP70 (designated AIHSP70) was cloned by the techniques of homological cloning and rapid amplification of cDNA end (RACE). The full length of AIHSP70 cDNA was 2651 bp in length, having a 5' untranslated region (UTR) of 96 bp, a 3' UTR of 575 bp, and an open reading frame (ORF) of 1980 bp encoding a polypeptide of 659 amino acids with an estimated molecular mass of 71.80 kDa and an estimated isoelectric point of 5.26. BLAST analysis revealed that the AIHSP70 gene shared high identity with other known HSP70 genes. Three classical HSP signature motifs were detected in AIHSP70 by InterPro, analysis. 3-D structural prediction of AIHSP70 showed that its N terminal ATPase activity domain and,C terminal substrate-binding domain shared high similarity with that in human heat shock protein 70. The results indicated that the AIHSP70 was a member of the heat shock protein 70 family. A semi-quantitive RT-PCR method was used to analyse the expression of AIHSP70 gene after the treatment of naphthalin which is one kind of polycyclic aromatic hydrocarbon (PAH) and the challenge of bacteria. mRNA expression of AIHSP70 in scallop was up-regulated significantly after the stimulation of naphthalin and increased with increasing naphthalin concentration. A clearly time-dependent expression pattern of AIHSP70 was observed after the scallops were infected by Vibrio anguillarum, and the mRNA expression reached a maximum level at 8 h and lasted to 16 h, and then dropped progressively. The results indicated that AIHSP70 could play an important role in mediating the environmental stress and immune response in scallop. (c) 2006 Elsevier Ltd. All rights reserved.

The first molluscan TCTP in Venerupis philippinarum: Molecular cloning and expression analysis

Translationally controlled tumor protein (TCTP) is one of the abundant and ubiquitously expressed proteins in metazoans In the present study, the first molluscan TCTP (denoted as VpTCTP) was identified from Venerupis philippinarum haemocytes by EST and RACE approaches The full-length cDNA of VpTCTP consisted of 1148 nucleotides with an open-reading frame of 555 bp encoding 184 amino acids The deduced amino acid sequence of VpTCTP shared high similarity with TCTPs from other species, indicating that VpTCTP should be a new member of TCTP family Several highly conserved motifs, including 5'terminal ologopyrimidine (5'TOP) starting sequence and rich AU and AUUT elements in 3'UTR, were also identified in VpTCTP The tissue and temporal expression of VpTCTP after Vi boo anguillarum challenge was recorded by quantitative real-time RT-PCR. VpTCTP transcript could be detected in all examined tissues with the highest expression level in haemocytes and the lowest in hepatopancreas Concerning the time-course expression in haemocytes, the relative expression of VpTCTP mRNA was down-regulated sharply from 6 h to 12 h post-infection. Then, the expression level was obviously up-regulated and reached 3.4-fold to that in the control group at 48 h post challenge As time progressed, the expression of VpTCTP recovered to the original level at 96 h. All these results indicated that VpTCTP was an acute-phase protein involved in the Immune response of V philippinarum (C) 2010 Elsevier Ltd. All rights reserved.

Molecular cloning, characterization and expression of a novel serine proteinase inhibitor gene in bay scallops (Argopecten irradians, Lamarck 1819)

Serine protease inhibitors, critical regulators of endogenous proteases, are found in all multicellular organisms and play crucial roles in host physiological and immunological effector mechanisms. The first mollusk serine proteinase inhibitor (designated AISPI) cDNA was obtained from the bay scallop Argopecten irradians by randomly sequencing a whole tissue cDNA library and rapid amplification of cDNA ends (RACE). The full-length cDNA of the scallop serine protease inhibitor was 1020 bp, consisting of a 5'-terminal untranslated region (UTR) of 39 bp, a 3'-terminal UTR of 147 bp with a canonical polyadenylation signal sequence AATAAA and a poly(A) tail, and an open reading frame of 834 bp. The AISPI cDNA encoded a polypeptide of 278 amino acids with a putative signal peptide of 22 amino acids and a mature protein of 256 amino acids. The deduced amino-acid sequence of AISPI contained six tandem and homologous domains similar to that of Kazal-type serine protease inhibitors, including the conserved sequence C-X(7)-C-X(6)-Y-X(3)-C-X(2,3)-C and six cysteine residues responsible for the formation of disulfide bridges, indicating that the AISPI protein from bay scallop should be a member of the Kazal-type serine protease inhibitor family. The temporal expression of AISPI was measured by semi-quantitative RT-PCR after injury or bacterial challenge. After the adductor muscle was wounded or injected with Vibrio anguillarum, the expression of AISPI mRNA in hemolymph was up-regulated and reached the maximum level at 8 and 16 h, respectively, and then progressively dropped back to the original level. The results indicated that AISPI could play an important role in injury healing and immune response in mollusks as it could be induced by injury and bacterial challenge. (c) 2005 Elsevier Ltd. All rights reserved.

Wangia profunda gen. nov., sp nov., a novel marine bacterium of the family Flavobacteriaceae isolated from southern Okinawa Trough deep-sea sediment

An orange-pigmented, Gram-negative, nonmotile, strictly aerobic and oxidase- and catalase-positive bacterium (SM-A87(T)) was isolated from the deep-sea sediment of the southern Okinawa Trough area. The main fatty acids were i15 : 0, i17 : 0 3OH, i15 : 1 G, i17 : 1 omega 9c, 15 : 0, i15 : 0 3OH and summed feature 3 (comprising i-15 : 0 2OH and/or 16 : 1 omega 7c). MK-6 was the predominant respiratory quinone. DNA G+C content was 35.8 mol%. Flexirubin-type pigments were absent. Phylogenetic analyses based on 16S rRNA gene sequences revealed that strain SM-A87(T) formed a distinct lineage within the family Flavobacteriaceae, with < 93% sequence similarity to the nearest strain of genus Salegentibacter. Moreover, strain SM-A87(T) could be distinguished from the nearest phylogenetic neighbors by a number of chemotaxonomic and phenotypic properties. On the basis of polyphasic analyses, it is proposed that strain SM-A87(T) be classified in a novel genus and a new species in the family Flavobacteriaceae, designated Wangia profunda gen. nov., sp. nov. The type strain is SM-A87(T) (CCTCC AB 206139(T)=DSM 18752).

中国海蓝子鱼科Family Siganidae分类和动物地理学特点

蓝子鱼科鱼类隶属于鲈形目、刺尾鱼亚目，广泛分布于我国东海、南海及台湾海域。本文根据中国科学院海洋研究所多年来在我国海域采集并收藏的以及部分补充采集的蓝子鱼标本，参考国际最新研究资料，对我国海域蓝子鱼科鱼类进行分类和动物地理学研究，共记录中国海域蓝子鱼科鱼类1属2亚属11种。经研究分析，得到如下结论： 1.中国沿海蓝子鱼科鱼类的多样性问题。国内曾有13种蓝子鱼的分布记录，其中蠕纹蓝子鱼Siganus vermiculatus和暗体蓝子鱼S. punctatissimus仅以前的学者做过名录形式的记载，并无标本收藏，本文中不做介绍。此外尖嘴蓝子鱼Siganus unimaculatus也可能存在于中国海域，有待于进一步研究。目前确定在中国海域有分布的蓝子鱼为11种，约占世界总种数（27）的40％多，种的多样性较高。 2. 形态学比较研究说明，蓝子鱼在体形、体色、牙齿、头骨、椎骨、耳石等特征方面存在种间差异，可作为蓝子鱼科鱼类分类的有效鉴别特征。对于外部形态极其相似的种，可借助内部解剖特征相辅助予以区分。 3．对蓝子鱼动物地理学特点研究分析结论如下：a.中国海域存在的11种蓝子鱼在印度－太平洋海域均有分布，没有地方特有种。b.长鳍蓝子鱼和褐蓝子鱼在我国东海、台湾海域和南海均有分布，褐蓝子鱼在黄海北部亦有分布，种群数量较大；其它9种，除凹吻蓝子鱼仅分布于南海外，均分布于南海及台湾海域，种群数量较小。c.中国海域蓝子鱼种类组成与邻近的菲律宾、马来西亚、印度尼西亚海域比较相似，中国分布的种类在这些海域均有分布，且分布于这一区域而中国没有记录的有5种。与日本相比，种类组成也比较相似，中日共有种达9种，仅分布于中国海域2种，仅分布于日本海域1种。 4. 作者参照国际最新研究结果和较为公认的分类系统Nelson（1994），将蓝子鱼科置于刺尾鱼亚目，而不再使用国内学者长期惯用的蓝子鱼亚目。蓝子鱼科包括一属蓝子鱼属Siganus Forsskål, 1775，两亚属：蓝子鱼亚属Siganus Forsskål, 1775 和罗蓝子鱼亚属 Lo Seale, 1906，同时使全部种名的变动和确定与国际最新研究结果(Woodland, 1990; 2001）取得一致。

中国海豆蟹科Family Pinnotheridae分类学研究

本文根据中国科学院海洋研究所收藏的豆蟹科标本和国内外相关文献，进行了中国海域豆蟹科的分类学研究。作者克服了豆蟹科分类难度大，研究积累较少，种的界限不清等困难，在短期内初步查清了中国海域豆蟹科种类及分布情况，共记述5亚科12属 40种。发现4新记录属，9新记录种，和豆蟹属1未定种Pinnotheres sp.，丰富了中国海豆蟹科区系内容，列出了共栖种中国海域全部记录过的宿主。 作者对鉴定过程中出现的疑难问题：“中型三强蟹Tritodynamia intermedia Shen, 1935是否为霍氏三强蟹 Tritodynamia horvathi Nobili, 1905的同物异名” 进行了细致比较、研究，否定了通行多年的Sakai（1976）的“T. intermedia Shen为T. horvathi Nobili的次异名”的结论，肯定了二者均为有效种。 对中国海域豆蟹科种类地理分布初步研究表明其分布与日本海域和东南亚海域都有相似之处，但有显著不同：仅发现于中国海域而未见于日本海域的豆蟹属Pinnotheres多达8种：涨腹豆蟹Pinnotheres excussus Dai et al., 1980，球豆蟹Pinnotheres pilulus Dai et al., 1980，锯颚豆蟹Pinnotheres serrignathus Shen, 1932，宽豆蟹Pinnotheres dilatatus Shen, 1932，青岛豆蟹Pinnotheres tsingtaoensis Shen, 1932，海阳豆蟹Pinnotheres haiyangensis Shen, 1932，钝颚豆蟹Pinnotheres obtusidentata（Dai et al., 1980），光豆蟹Pinnotheres luminatus Dai et al., 1980）；仅发现于中国海域的三强蟹属Tritodynamia有4种：福建三强蟹Tritodynamia fujianensis Chen, 1979，长腿三强蟹Tritodynamia longipropodum Dai et al., 1980，宽身三强蟹 Tritodynamia dilatatum Yang et Sun,1996和海南三强蟹 Tritodynamia hainanensis Dai et al.g, 1980；而未发现于中国海域的日本特有种多达13种。结果表明这一类群由于共栖和地域分化程度较高乃至地区性特有种数显著较多。 论文通过对豆蟹科系统分类和初步的动物地理学研究，搞清了中国海豆蟹科的种和分布、多样性及其宿主等基本情况。为今后研究豆蟹科种类生物学、生态学特性提供了基础资料。

Two new species of Family Prasiolaceae (Prasiolales, Chlorophyta) from China Sea

Amomg the 30 known species of the algal family Prasiolaceae (Prasiolales, Chlorophyta), nine marine species have been found in marine environments but none in China seas. We reported here two new species Prasiola fangchengensis Luan et Ding sp. nov. and Prasiola volcanica Luan et Ding sp. nov. from subtropical coastal water of southern China.

First record of the family Spengeliidae (Hemichordata : Enteropneusta) from Chinese waters, with description of a new species

Six species belonging to two families of Hemichordata have previously been recorded in Chinese waters. This paper records the discovery and description of a new species of the genus Glandiceps found in Jiaozhou Bay, Qingdao, Shandong Province, named Glandiceps qingdaoensis. The new species has a long proboscis with dorsal and ventral grooves, a stomochord with a long vermiform process, a proboscis cavity with a dorsal median, right and left glomeruli, right and left glomeruli very large and encircling the stomochord, a proboscis skeleton in the cavity extends into the median posterior of the collar, a well-developed dorsal ventral muscular septum in the proboscis cavity dividing the cavity completely into two separate parts. The collar cord is without giant nerve roots. The trunk with four distinct regions that can be recognized externally: branchial-genital region, genital region, hepatic region, and intestinal region. The dorsal pharynx is large and the gill pores are small. The tongue bars are encircled by vesicles, and the first gonad commences at the level of the second or third gill slit.

The Apertospathulidae, a new family of haptorid ciliates (Protozoa, Ciliophora)

We studied the morphology of three rare haptorid ciliates, using live observation and silver impregnation: Apertospathula verruculifera n. sp., Longispatha elegans n. gen., n. sp., and Rhinothrix porculus (Penard, 1922) n. gen., n. comb. Simple ethanol fixation (50-70%, v/v) is recommended to reveal the ciliary pattern of "difficult" ciliates, such as R. porculus, by protargol impregnation. The three genera investigated have a distinct feature in common, viz., a lasso-shaped oral bulge and circumoral kinety, where the right half is slightly to distinctly longer than the left and the circumoral kinety is open ventrally. Thus, they are united in a new spathidiid family, the Apertospathulidae n. fam., which probably evolved from a Bryophyllum-like ancestor by partial reduction of the oral bulge and circumoral kinety. Apertospathula verruculifera has a wart-like process, the palpus dorsalis, at the anterior end of the dorsal brush. The right branch of the circumoral kinety is only slightly longer than the left one. Longispatha elegans has a straight oral bulge and circumoral kinety, the right branch of which extends to the posterior end of the body while the left branch ends in the anterior third of the body. Rhinothrix porculus, a curious ciliate with a snout-like dorsal elongation of the oral bulge, the palpus oralis, has a highly characteristic ciliary pattern: the oral pattern is as in Longispatha, but the bulge and circumoral kinety extend spirally to the posterior end of the body while the somatic kinetics course meridionally. This is achieved by inserting some shortened kinetics in the curves of the oral bulge.

Taxonomic study on the family Mitridae (Mollusca : Gastropoda : Neogastropoda) from the China's Seas. Subfamilies: Imbricariinae and Cylindromitrinae

Ninteen species of subfamilies Imbricariinae and Cylindromitrinae, family Mitridae, are recorded from the China's seas. Of which, one genus and six species are recorded for the first time from China's seas, i.e., genus Ziba Adams H and Adams A, Cancilla (Cancilla) carnicolor, Ziba duplilirata, Z. insculpta, Neocancilla circula, Scabricola (Scabricola) desetangsii, Scabricola (Swainsonia) ocellata ocellata.