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拟穴青蟹E93基因的鉴定及表达调控分析

海洋渔业 2024年第3期46卷 307-315页

作者：宋诗颖张凤英王伟刘志强马凌波赵明上海海洋大学水产与生命学院上海201306 中国水产科学研究院东海水产研究所农业农村部远洋与极地渔业创新重点实验室上海200090

为研究E93在拟穴青蟹中的功能和调控特征,利用转录组测序和分子克隆鉴定获得了拟穴青蟹(Scylla paramamosain)的E93基因,命名为Sp-E93,并对其基本生物信息、系统进化、组织分布以及激素调控的模式进行了分析。结果发现,Sp-E93的mRNA长度... 详细信息

为研究E93在拟穴青蟹中的功能和调控特征,利用转录组测序和分子克隆鉴定获得了拟穴青蟹(Scylla paramamosain)的E93基因,命名为Sp-E93,并对其基本生物信息、系统进化、组织分布以及激素调控的模式进行了分析。结果发现,Sp-E93的mRNA长度为3756 bp,包含2982 bp开放阅读框(ORF),编码993个氨基酸,预测蛋白质的相对分子质量为106.34 kDa,含有两个保守的helix-turn-helix(HTH)结构域,而HTH结构域外的其他区域序列在不同物种中一致度较低。系统进化分析发现,该基因的进化与物种进化基本一致。Sp-E93在拟穴青蟹成体的Y器官和大颚器中表达量最高,且在雄蟹中的表达显著高于雌蟹(P<0.05)。此外,甲基法尼酯和甲氧普林能显著上调雌蟹肝胰腺中E93基因的表达,但对卵巢中E93的表达无调控作用;20E可以调控肝胰腺E93基因的表达,但对卵巢中E93的表达无显著调控。结果表明,青蟹的E93基因在拟穴青蟹的激素信号中发挥作用,但调控模式较昆虫发生了一定进化,研究结果可为进一步研究甲壳动物E93的功能奠定基础。

关键词：拟穴青蟹 20E 甲基法尼酯 E93 基因表达调控

Sensitive and rapid detection of two toxic microalgae Alexandrium by loop-mediated isothermal amplification

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Acta Oceanologica Sinica 2012年第2期31卷 139-146页

作者： ZHANG Fengying SHI Yanhong JIANG Keji XU Zhaoli ma lingbo East China Sea Fisheries Research Institute Chinese Academy of Fishery SciencesShanghai 200090China Key and Open Laborutory of marine and Estuarine Fisheries Ministry of Agriculture of ChinaShanghai 200090China

A loop-mediated isothermal amplification （LAMP） assay was designed and evaluated for rapid de- tection of the toxic microalgae Alexandrium catenella and A. minutum, which can produce paralytic shellfish poisoning （PSP）. Two sets of four specific primers targeting these two species were derived from the sequence of internal transcribed spacer （ITS） of ribosomal DNA. The method worked well in less than an hour under isothermal conditions of 65℃. LAMP specificity was validated in closely related algae as a comparison, suggesting the strict specificity of the LAMP primers. Two visual inspection approaches were feasible to interpret the positive or negative results. The detection lim- its of A. catenella and A. minutum samples using the LAMP assay were found to be 5.6 and 4.5 pg DNA, respectively. The sensitivity of this LAMP assay was 10 or 100-fold higher than Polymerase Chain Reaction （PCR） method in detecting the two microalgae. These characteristics of species specificity, sensitivity, and rapidity suggest that this method has the potentiality in the monitoring of red tide caused by A. catenella and A. minutum.

关键词： Alexandrium eatenella Alexandrium minutum detection loop-mediated isothermal amplification (LAMP) ribosomal DNA internal transcribed spacer (ITS)

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不同饵料对拟穴青蟹大眼幼体变态发育的影响

水产科学 2024年第3期43卷 429-436页

作者：乔光德马春艳张凤英王伟赵明谌微傅愔刘志强马凌波上海海洋大学水产与生命学院上海201306 中国水产科学研究院东海水产研究所遗传育种与生物技术试验室上海200090

为探寻适用于拟穴青蟹大眼幼体发育时期投喂的饵料,在自然光照、水温(30±1)℃、盐度23条件下,将(1000±30)只大眼幼体放入盛水360 L的孵化桶中孵化,投喂卤虫无节幼体、冰冻桡足类、冰冻卤虫、配合饲料,及不投喂任何饵料,直至蜕壳变态... 详细信息

为探寻适用于拟穴青蟹大眼幼体发育时期投喂的饵料,在自然光照、水温(30±1)℃、盐度23条件下,将(1000±30)只大眼幼体放入盛水360 L的孵化桶中孵化,投喂卤虫无节幼体、冰冻桡足类、冰冻卤虫、配合饲料,及不投喂任何饵料,直至蜕壳变态为仔蟹,并分析不同饵料投喂对养殖水体的水质、大眼幼体蜕壳变态、仔蟹免疫相关酶及消化酶活性的影响等。结果显示:投喂卤虫无节幼体组的养殖水体中氨氮、亚硝态氮含量最低,成活率最高,幼体发育时间比其他3个投喂组缩短1~3 d,甲长、甲宽均显著大于配合饲料组的个体,但与冰冻桡足类组、冰冻卤虫组无显著差异;4个投喂组间仔蟹酸性磷酸酶、碱性磷酸酶、超氧化物歧化酶、α-淀粉酶活性无显著差异;卤虫无节幼体组溶菌酶活性、总抗氧化能力与冰冻桡足类、冰冻卤虫组无显著差异,但显著高于配合饲料组;卤虫无节幼体组胰蛋白酶活性和脂肪酶活性显著高于其他3个投喂组。试验结果表明,卤虫无节幼体对育苗水体水质及仔蟹的蜕壳成活率、生长、免疫相关酶活性、消化酶活性均存在积极作用,更适合作为拟穴青蟹大眼幼体阶段的饵料;冰冻桡足类、冰冻卤虫使用效果稍差,但均优于配合饲料组,可以用于替代部分卤虫无节幼体,以降低育苗成本,简化生产流程。试验结果可为拟穴青蟹苗种人工培育过程中饵料选择提供依据。

关键词：饵料拟穴青蟹大眼幼体变态发育

基于岩石物理模型的碳酸盐岩物性参数替换方法

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CT理论与应用研究（中英文） 2024年第3期33卷 273-288页

作者：马凌波张佳佳张广智谷一鹏中国石油大学(华东)地球科学与技术学院山东青岛266000

碳酸盐岩的物性参数在油气勘探和开发的过程中起着重要的作用,且碳酸盐岩为具有多重孔隙类型的岩石,复杂的孔隙类型使得孔隙度与弹性参数之间的关系非常离散。本文基于岩石物理模型,提出一种碳酸盐岩物性参数替换的方法,首先对碳酸盐岩... 详细信息

碳酸盐岩的物性参数在油气勘探和开发的过程中起着重要的作用,且碳酸盐岩为具有多重孔隙类型的岩石,复杂的孔隙类型使得孔隙度与弹性参数之间的关系非常离散。本文基于岩石物理模型,提出一种碳酸盐岩物性参数替换的方法,首先对碳酸盐岩储层进行岩石物理建模,对模型中的等效孔隙纵横比进行反演,在进行替换时,保持其他参数不变,只改变孔隙度、方解石含量、含水饱和度和孔隙形状的体积分数中的一项。再结合AVO理论,进行正演模拟,正演模拟揭示孔隙度与孔隙形状的变化对地震响应的影响更为强烈,远远大于方解石含量、含水饱和度变化的影响,方解石含量和含水饱和度的变化对于地震响应的影响较微弱。实际资料应用表明,文章提出的碳酸盐岩物性参数的替换方法可以有效地分析物性参数及孔隙形状变化的影响,表征岩石的物理性质并判断岩石的孔隙类型。

关键词：岩石物理反演碳酸盐岩孔隙结构物性参数

Identification and profiling of microRNAs of Euphausia superba using Illumina deep sequencing

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Journal of Oceanology and Limnology 2018年第6期36卷 2278-2287页

作者： SONG Wei ZHU Yefei WANG Lumin JIANG Keji ZHANG Fengying ma Chunyan ma lingbo Key Laboratory of Oceanic and Polar Fisheries Ministry of AgricultureEast China Sea Fisheries Research InstituteChinese Academy of Fishery Sciences Key Laboratory of East China Sea Fishery Resources Exploitation Ministry of AgricultureEast China Sea Fisheries Research InstituteChinese Academy of Fishery Sciences

MicroRNAs(miRNAs) are an abundant class of conserved, non-coding small RNAs that play important role in gene regulation at post translational level. There have been no reports on the miRNAs of the Antarctic krill E uphausia superba despite the species' crucial position in Antarctic food webs. Two small RNA libraries were constructed from eyestalk and muscle, subsequently, and deep sequencing analysis was performed to investigate and profile E. superba miRNAs. A total of 19 304 586 and 23 005 104 unique sequences were obtained from the eyestalk and muscle, respectively. After compared the small RNA sequences with the Rfam database, 12 342 039 and 7 907 477 reads in eyestalk and muscle were matched to the transcriptome sequence of E. superba. A total of 236 distinct miRNAs were identified after annotation to known animal miRNAs registered in miRBase 21. In both libraries, the most abundant known miRNA were miR-750 with 92 583 reads in muscle and miR-1304-3 p with 56 386 reads in eyestalk while the average value was less than 106, revealing a wide range of different expression levels. In addition, miR-277 a enriched in both libraries and may be involved in modulating Krebs cycle by targeting to Vimar. Differential expression analysis showed that 56 mature miRNAs were significantly up/down regulated according to expression fold change. It appeared that the expression of several abundant miRNAs maybe tissue-specific or tissue-bias. Notably, the expression pattern of miR-750 and miR-1 family, which was suggested as the crucial candidates, involved in muscle development. Taken together, this study provides the first miRNA profile of E. superba and some of these miRNAs were expected to play important role in immune response, reproduction, energy metabolism, and muscle development and so on. Thus, the results provides a reference for functional studies of miRNAs in E. superba.

关键词： microRNA Antarctic krill Illumina deep-sequencing eyestalk muscle

The Taxonomic Status of Gymnura bimaculata and G. japonica： Evidence from Mitochondrial DNA Sequences

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Journal of Life Sciences 2012年第1期6卷 9-13页

作者： Anglv Shen Chunyan ma Yong Ni Zhaoli Xu lingbo ma East China Sea Fisheries Research Institute Chinese Academy of Fishery Sciences Shanghai 200090 China

Japanese butterfly ray Gymnura japonica from twinspot butterfly ray G. bimaculata based on a pair of white spots behind eyes or not, which was not reliable. To clarify the taxonomic status of G. japonica and G. bimaculata, the nucleotide variation between the two butterfly rays was examined using mitochondrial DNA sequence comparisons. Approximately 585 bp of 16S ribosomal RNA （rRNA） and 1,128 bp cytochrome b （cyt b） genes were sequenced from representatives of two butterfly rays species in East China Sea. The results showed that there were the same sequences of 16S rRNA gene between two butterfly rays; six sites were variable among two butterfly rays of cyt b genes, the proportion of polymorphie loci was 0.53%, and two haplotypes were defined which genetic distance was 0.5%. Combined with the morphological character and the analysis of mtDNA sequence indicated that twinspot butterfly ray G. bimaculata was a synonym of Japanese butterfly ray G. japonica.

关键词： 16S rRNA cyt b morphological character taxonomic status butterfly ray.

The Novel Hatchery Facilities Based on main Effect Factors of Seedling Rearing of Mud Crab （Scylla spp.） in China

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Journal of Life Sciences 2010年第3期4卷 36-42页

作者： Zhenguo Qiao Jiangang Wang Zhongli Yux Keji Jiang lingbo ma Key Laboratory of marine and Estuarine Fisheries Resources and Ecology of Ministry of Agriculture of China East China SeaFisheries Research Institute Chinese Academy of Fishery Sciences Shanghai 200090 China Shanghai Fengxian Municipal Center for Fishery Technology Promotion Shanghai 201400 China

Mud crab is an important economic crab cultured in Pacific and Indian Ocean Rim. Based on the knowledge about the characteristic requirements in seedling cultivation of mud crab under high-temperature conditions, this article reviews and discusses major factors affecting seedling rearing of mud crabs, and presents novel hatchery facilities for improvement of seedling rearing. The results of our study showed that major factors affecting seedling cultivation of mud crabs included：（1） the micro-ecological environment and characteristics of water quality in seedling rearing ponds, and a double-bottom cultivation facilities and cultivation techniques that meet the requirement; （2） salinity, temperature, light, NOE-N, NH4-N and other physical and chemical environmental factors that affect survival probability ofzoaea; （3） the total number of bacteria in the zoaea-culturing pond and the number of vibrio in seedling rearing; and （4） nutritional intensification with bait organisms in zoaea-culturing. To meet and improve these conditions, a novel hatchery facility was designed to modify the pond roof in accordance with the characteristics of the zoaea stage, and improve natural light thermostat control in seedling cultivation of mud crabs.

关键词： Seedling rearing Scylla novel hatchery facilities nutrition micro-ecological environment physical and chemical environment.

The research progress on food organism culture and technology utilization in crab seed production in ponds in China

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Agricultural Sciences 2013年第10期4卷 563-569页

作者： Jibing Qi Xiaolian Gu lingbo ma Zhenguo Qiao Kai Chen East China Sea Fishery Research Institute Chinese Academy of Fishery Science Shanghai China Institute of Aquaculture and Life Shanghai Ocean University Shanghai China

Eriocheirsinensis, Portunus trituberculatus and Scylla paramamosain are important commercial culture crab species in China. Traditional factory breeding of crabs depends on Artemia nauplius. The rising price of Artemia cysts has led to the decline of the economic benefit of the crab breeding factory. Factory crab breeding has been gradually replaced by pond breeding in recent years. E. sinensis and P. trituberculatus have been bred mainly in ponds. Meanwhile, S. paramamosain is still mainly bred in factories because of the crudeness of pond breeding. The research progress on food organism utilization in the three species of commercially bred crabs was reviewed in this paper. In the workshop seeds production, rotifer and Artemia nauplii were necessary in the early stages from zoea I to zoea II in the three crab species. Adult artemia, minced fish and shellfish were fed to the larvae in the later zoea stages from zoea III to Megalopa.?The rising of the price of artemia eggs made people find other feed organism to replace artemia. Copepods have been used in crab seeds production in pond from zoea III stage to replace artemia in recent years, which has reduced the cost of seeds production.

关键词： Food Organisms Crab Seeds Ponds

维生素E与熊去氧胆酸对非酒精性脂肪肝大鼠的协同肝保护机制

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基因组学与应用生物学 2019年第12期38卷 5713-5718页

作者：李微高强马凌波哈尔滨医科大学附属第二医院

非酒精性脂肪性肝病(NAFLD)是全世界最常见的肝病,而非酒精性脂肪肝炎(NASH)是其中一种,具有潜在的严重性,其发展涉及多次打击过程。一些临床研究表明,熊去氧胆酸(UDCA)和维生素E单独或联用对治疗NASH患者有明显的改善作用,但效果具有... 详细信息

非酒精性脂肪性肝病(NAFLD)是全世界最常见的肝病,而非酒精性脂肪肝炎(NASH)是其中一种,具有潜在的严重性,其发展涉及多次打击过程。一些临床研究表明,熊去氧胆酸(UDCA)和维生素E单独或联用对治疗NASH患者有明显的改善作用,但效果具有一定可逆性。而关于UDCA和维生素E联用治疗NASH的机制研究也尚少,因此本研究希望通过探究UDCA与维生素E联用对改善NASH大鼠的效果与机制,为NASH的治疗提供更多的理论依据。构建NASH大鼠模型,通过喂养不同饮食分别设立对照组,模型组和2组给药治疗组(UDCA单独用药;UDCA/VitE联合用药),12周后对大鼠肝组织进行病理分析,检测AST、ALT、MDA、SOD等生化指标和组织NF-κB、TNF-α蛋白表达水平,对比不同组间差异,评估给药治疗的疗效,分析可能的机制;UDCA能有效改善NASH的各种组织特征或生化指标,并且与维生素E一起对NASH大鼠的肝保护具有协同作用,这种协同作用可能与抑制肝细胞脂肪变性和炎细胞浸润,减轻肝损伤和胰岛素抵抗有关。

关键词：维生素E 熊去氧胆酸非酒精性脂肪肝协同肝保护