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Bacterial and cyanobacterial diversities determined by T-RFLP analyses in the Jiaozhou Bay

Acta Oceanologica Sinica 2006年第4期25卷 113-123页

作者： REN Jing DANG Hongyue song linsheng SUN song AN Liguo College of Life Science Shandong Normal University Jinan 250014 China Institute of Oceanology Chinese Academy of Sciences Qingdao 266071 China

The methods of DAPI staining epifluorescence microscopy and T-RFLP analysis were used to analyze the microbial abundance and diversity in surface seawater sampled from 12 stations inside and outside of the Jiaozhou Bay during a survey on 12 and 13 September 2004. The abundance of total microbes is in the range of 10^6～ 10^7 cells/cm^3, similar to those of most semi-enclosed bays in the temperate areas in the world. The highest microbial densities occur in the northeastern part of the Jiaozhou Bay, around the mouths of Loushan and Licun Rivers and the Hongdao aquacultural farming areas, suggesting that the degree and characteristics of pollutions, along with geographical and hydrological effects, may be important determinants affecting the abundance and distribution of bacteria in the Jiaozhou Bay. Bacterial communities inside and outside of the Jiaozhou Bay can be grouped into three classes based on T-RFLP patterns and cluster analyses. Stations at the water channel of the bay mouth and outside, such as D1, D3, D5, D6 and D7, are grouped together to stand for the outside bacterial community interacting with the environment outside of the Jiaozhou Bay. Stations of the innermost side of the Jiaozhou Bay, such as A3, A5, B2 and Y1, are grouped together to stand for the residential bacteria community. Stations C1, C3 and CA are grouped together and may stand for the transitional bacterial assemblage between the residential community and the outside community. However, there is no such a defined relationship for the case of cyanobacterial diversity, indicating the fact that cyanobacteria are more flexible and adaptable to all kinds of conditions.

关键词： bacterial diversity cyanobacteria 16S rDNA T-RFLP Jiaozhou Bay

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Molecular cloning and expression analysis of interleukin - 1β from Japanese sea perch(Lateolabrax japonicus )

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Acta Oceanologica Sinica 2006年第1期25卷 127-136页

作者： QIU Lihua song linsheng WU Longtao CAI Zhonghua JIANG Shigui Institute of Oceanology Chinese Academy of Sciences Qingdao 266071 China South China Sea Fisheries Research Institute Chinese Academy of Fishery Sciences Guangzhou 510300 China Graduate School Chinese Academy of Sciences Beijing 100039 China

The technology of homology cloning and anchored PCR was used to clone the IL-1β gene from the Japanese sea perch （Lateolabrax iaponicus）. The full-length cDNA of sea perch IL-1β was 1 310 bp, including a 5＇ untranslated regiop （UTR） of 136 bp, a 3＇ UTR ot 430 bp, and an ORF of 774 bp encoding a polypeptide of 258 amino acids with an estimated molecular mass of 29.31 kDa. The searches for nucleotides and protein sequence similarities with the BLAST analysis indicated that the deduced amino acid sequence of sea perch IL-1β was homological to the IL-1β in other fish species and even the mammalian. Conserved signature sequences of the IL-1β gene family were found in the sea perch IL-1β deduced amino acid sequence. Temporal expressions of the IL-1β gene in LPS or iridovirus challenged group and in control group were measured by the semi-quantitative RT-PCR. The mRNA transcripts of IL-1β could be detected in head-kidney, spleen, liver, gill and heart of the healthy individuals, and the expression level of IL-1β in head-kidney, spleen and gill was higher than that in liver and heart, but it was hard to be detected in the brain. After being stimulated by the LPS or iridovirus, the IL-1β expression in most of examined tissues was up-regulated, and also could be detected in the brain. These results indicated that the expression of sea perch IL-1β was constitutive and could be up-regulated by immune effector stimulation. Therefore the sea perch IL-1β could play a critical role in the host-pathogen interaction.

关键词： Japanese sea perch interleukin-1β gene cloning cDNA mRNA expression

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Screening microsatellite markers from EST sequences of bay scallop Argopecten irradians

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High Technology Letters 2006年第1期12卷 97-102页

作者：王玲玲 song linsheng Xu Wei Zhang Huan Institute of Oceanology Chinese Academy of Sciences Qingdao 266071 P.R. China Graduate School Chinese Academy of Sciences Beijing 100039 P.R. China

Seventy-five simple sequence repeats （SSRs） were identified by the bioinformatic analysis from 5008 expressed sequence tags （ESTs） of Argopecten irradians. Among the SSRs, the number of repeat nucleotide varied from 2 to 6. Dinucleotide and trinueleotide repeat motifs were dominant in EST-SSRs of bay scallop, with a proportion of 80% over the total screened SSRs. Twenty-nine pairs of primer were designed based on the flank sequences of the selected ESTs using the software of Primet 5, and verified under the given PCR reaction condition. Eighteen of the 29 primer pairs resulted in the expected products, while the remaining either failed to produce any fragments or yielded products over expected size. Thirteen of the 18 SSRs, accounting for 72%, were detected to show polymorphism in the examined scallop samples. A preliminary test in this study indicated that the majority of the identified SSRs were informative in the cultured bay scallops, making them suitable for the population and other genetic analysis. EST-SSR markers have more advantages than the traditional genomic-derived SSRs and there is a wide range of application in comparative mapping, functional gene cloning and marker assisted selection. This research provides a reference to the identification of EST-SSRs with relative bioinformatic analysis from aquaculture species, as well as to those with a large number of ESTs.

关键词： Argopecten irradians microsatellite markers expressed sequence tags

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Spatial compartmentalization and temporal stability of associated microbiota in Pacific oyster Crassostrea gigas

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Journal of Oceanology and Limnology 2024年第4期42卷 1348-1358页

作者： Qiang FU Zichao YU Junyan ZHAO Lei GAO Ning KONG Lingling WANG linsheng song Liaoning Key Laboratory of Marine Animal Immunology and Disease Control Dalian Ocean UniversityDalian 116023China Functional Laboratory of Marine Fisheries Science and Food Production Processes Qingdao National Laboratory for Marine Science and TechnologyQingdao 266237China Southern Marine Science and Engineering Guangdong Laboratory(Zhuhai) Zhuhai 519000China Dalian Key Laboratory of Aquatic Animal Disease Prevention and Control Dalian Ocean UniversityDalian 116023China

The Pacific oyster Crassostrea gigas,one of the most exploited molluscs in the world,has suffered from massive mortality in recent decades,and the occurrence mechanisms have not been well *** this study,to reveal the relationship of associated microbiota to the fitness of oysters,temporal dynamics of microbiota in the gill,hemolymph,and hepatopancreas of *** during April 2018-January 2019 were investigated by 16 S rRNA gene *** microbiota in *** exhibited tissue heterogeneity,of which Spirochaetaceae was dominant in the gill and hemolymph while Mycoplasmataceae enriched in the ***-occurrence network demonstrated that the gill microbiota exhibited higher inter-taxon connectivity while the hemolymph microbiota had more *** richness(Chao 1 index)and diversity(Shannon index)of microbial community in each tissue showed no significant seasonal variations,except for the hepatopancreas having a higher richness in the ***,beta diversity analysis indicated a relatively stable microbiota in each tissue during the sampling period,showing relative abundance of the dominant taxa exhibiting temporal *** indicate that the microbial community in *** showed a tissue-specific stability with temporal dynamics in the composition,which might be essential for the tissue functioning and environmental adaption in *** work provides a baseline microbiota in *** and is helpful for the understanding of host-microbiota interaction in oysters.

关键词： Pacific oyster associated microbiota spatial compartmentalization temporal stability

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我国海水养殖贝类病害发生现状及防控策略

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水产学报 2023年第11期47卷 148-152页

作者：宋林生大连海洋大学辽宁省海洋动物免疫学与疫病防控重点实验室

贝类养殖是我国水产养殖业的重要组成部分，在促进经济社会发展、提高就业率、改善人民生活及保护生态环境等方面发挥了重要作用，其经济、社会和生态功能日益凸显。近年来，因环境胁迫和病原侵染引发的海水养殖贝类暴发性病害时有发生... 详细信息

贝类养殖是我国水产养殖业的重要组成部分，在促进经济社会发展、提高就业率、改善人民生活及保护生态环境等方面发挥了重要作用，其经济、社会和生态功能日益凸显。近年来，因环境胁迫和病原侵染引发的海水养殖贝类暴发性病害时有发生，所造成的经济损失巨大，严重阻碍了海水养殖产业的绿色高质量发展。贝类缺乏适应性免疫，且主要养殖于开放或半开放水体，难以通过疫苗注射或环境调控等手段进行病害防控。贯彻“绿色防控”的理念，加快病害预警预报体系建设是海水养殖贝类病害防控的重要发展方向。本文综述了我国海水贝类养殖及病害发生的现状，并结合团队前期研究和病害预警预报工作实践,提出了海水养殖贝类病害防控策略，以期提升我国水产动物疫病防控能力，为保障水产品稳定安全供给、促进水产养殖业绿色高质量发展提供参考。

关键词：海水养殖贝类病害发生病害防控预警预报

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海水养殖贝类病害预警预报技术及其应用

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大连海洋大学学报 2020年第1期35卷 1-9页

作者：宋林生大连海洋大学辽宁省海洋动物免疫学与疫病防控重点实验室辽宁省海洋动物免疫学重点实验室

贝类是中国海水养殖产业的重要组成部分,近年来,海水养殖贝类病害时有发生,尤其在夏季常出现大规模死亡,造成重大经济损失,严重威胁了贝类养殖产业的健康发展。由于贝类缺乏适应性免疫,且通常养殖于开放水体,难以通过药物投放、注射或... 详细信息

贝类是中国海水养殖产业的重要组成部分,近年来,海水养殖贝类病害时有发生,尤其在夏季常出现大规模死亡,造成重大经济损失,严重威胁了贝类养殖产业的健康发展。由于贝类缺乏适应性免疫,且通常养殖于开放水体,难以通过药物投放、注射或水质调节等措施控制病害的发生,因此,建立养殖贝类健康评价体系,加强养殖生态环境监测及病害发生的预警预报,已成为保障中国贝类养殖业健康可持续发展、确保粮食和生态安全的迫切需求。本文从病原、宿主、环境间的耦合关系出发,结合团队前期研究和工作实践,综述了水产养殖动物病害预警预报研究进展,同时针对北黄海和渤海贝类养殖海区的生态系统特征及贝类养殖现状,并结合病害预警预报的工作实践,重点阐述了海水养殖贝类病害预警模型构建的思路、原理和初步应用,以期为建立海水养殖动物病害防控体系、保障中国海水养殖业的绿色健康发展提供理论参考和技术支撑。

关键词：养殖贝类病害发生预警技术应用前景

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Phylogenetic relationships of five species of Dorippinae (Crustacea, Decapoda) revealed by 16S rDNA sequence analysis

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Acta Oceanologica Sinica 2004年第3期23卷 513-519页

作者： FANYu LIXinzheng songlinsheng CAIZhonghua InstituteofOceanology ChineseAcademyofSciencesQingdao266071China

A molecular phytogeny is presented for the subfamily Dorippinae (including 9 individuals, representing 5 species and 4 genera), based on the sequence data from 16S rRNA gene. Two-cluster test between lineages in these phylogenetic trees has been performed. On the basis of rate constancy, the rate of nucleotide substitutions of 16S rDNA sequence data is estimated as 0.27% per million years. The analysis strongly supports the recognition of the Dorippinae as a monophyletic subfamily. Phylogenetic tree indicates that the subfamily Dorippinae is divided into two main clades, and genus Dorippe appears basal in the subfamily, diverging from other species 36.6 Ma ago. It is also clear that the Heikea is closely related to the genus Neodorippe. The divergence time between them is 15.8 Ma.

关键词： crab Dorippinae Dorippe tenuipes Heikea japonica Neodorippe callida Paradorippe granulata Paradorippe polita 16S rDNA phytogeny divergence time

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Identification and cloning of the genes involved in immuine defensefrom scallop

Identification and cloning of the genes involved in immuine ...

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贝类学会第七次会员代表大会暨第十一次学术讨论会

作者： linsheng song Institute of Oceanology, the Chinese Academy of Sciences, Qingdao 266071, China

＜正＞ Scallop aquaculture in China has beeh growing rapidly in the past decades and there is a big scallop culture industry which plays an important role in the economic development of the coastal provinces in China. Chlamys farreri and Argopecten irradians are the important cultured scallop species which are cultured widely in the northern coastal provinces. Recently, scallop culture in China is experiencing from a mortality problem, and suffering from a great economic loss.

关键词： scallop EST Gene cloning Immune defense Gene expression

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海湾扇贝g型溶菌酶基因cDNA的克隆与分析

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高技术通讯 2005年第7期15卷 101-106页

作者：邹慧斌宋林生胥炜杨官品中国科学院海洋研究所实验海洋生物学开放实验室青岛26607中国海洋大学海洋生命学院青岛266003 中国科学院海洋研究所实验海洋生物学开放实验室青岛266071 中国海洋大学海洋生命学院青岛266003

通过EST和锚定PCR技术,从海湾扇贝(Argopecten irradians)中克隆得到一种溶菌酶基因的全长cDNA(全长659bp,编码200个氨基酸).BLASTx分析的结果显示,它和脊椎动物g型溶菌酶相似性较高,却不同于无脊椎动物中已发现的c型和i型溶菌酶.在其... 详细信息

通过EST和锚定PCR技术,从海湾扇贝(Argopecten irradians)中克隆得到一种溶菌酶基因的全长cDNA(全长659bp,编码200个氨基酸).BLASTx分析的结果显示,它和脊椎动物g型溶菌酶相似性较高,却不同于无脊椎动物中已发现的c型和i型溶菌酶.在其编码的氨基酸序列中发现了g型溶菌酶的活性中心(Glu 82, Asp 97, Asp 108),同时6个保守的半胱氨酸也与鸟类和鱼类的g型溶菌酶相一致.结合BLASTX分析的结果,可以确认所获得的cDNA序列是一种在无脊椎动物中首次发现的g型溶菌酶的编码序列.采用Clustalw 软件,对多种脊椎动物c型、g型溶菌酶和无脊椎动物c型、i型溶菌酶以及本研究发现的无脊椎动物g型溶菌酶进行了分析,结果发现,无脊椎动物c型、i型溶菌酶和脊椎动物c型溶菌酶同源性较高,而海湾扇贝g型溶菌酶和脊椎动物g型溶菌酶同源性较高.由此说明c型、g型溶菌酶从无脊椎到脊椎动物的进化是平行发生的,这对进一步研究溶菌酶及其分子进化具有重要意义.

关键词：海湾扇贝基因cDNA g型克隆无脊椎动物全长cDNA cDNA序列 PCR技术溶菌酶基因氨基酸序列活性中心半胱氨酸编码序列首次发现分子进化同源性 c型 EST 相似性 i型鱼类鸟类

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贝类炎症及其发生机制研究进展

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大连海洋大学学报 2023年第3期38卷 369-379页

作者：孙洁洁宋林生大连海洋大学辽宁省海洋动物免疫学与疫病防控重点实验室辽宁省海洋动物免疫学重点实验室辽宁大连116023

贝类在中国海水养殖业中占据重要地位,养殖贝类病害的频发,严重阻碍了贝类养殖业的可持续健康发展。炎症是机体对刺激的一种防御反应,在抵御病原菌感染中发挥重要作用,但过度的炎症反应会造成机体组织损伤,严重时会引发个体死亡。目前,... 详细信息

贝类在中国海水养殖业中占据重要地位,养殖贝类病害的频发,严重阻碍了贝类养殖业的可持续健康发展。炎症是机体对刺激的一种防御反应,在抵御病原菌感染中发挥重要作用,但过度的炎症反应会造成机体组织损伤,严重时会引发个体死亡。目前,已经在贝类中观察到了炎症现象,并鉴定出多种炎症细胞因子和炎症相关信号通路。本文围绕贝类炎症细胞因子种类与生物学功能、炎症相关信号通路及炎症反应的形态学特征,总结了近年来贝类炎症研究进展,重点阐述了炎症发生的分子机制,并针对研究中存在的问题,提出不同类型炎症细胞因子的挖掘、炎症细胞因子的免疫学功能及其与不同类型免疫细胞之间的关系等是该领域未来的重点研究方向,以期为推动贝类养殖产业绿色高质量发展提供科学参考。

关键词：贝类炎症反应炎症细胞因子信号转到免疫调节

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