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Validation of reference genes for gene expression studies in the dinoflagellate Akashiwo sanguinea by quantitative real-time RT-PCR

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Acta Oceanologica Sinica 2016年第8期35卷 106-112页

作者： DENG Yunyan HU Zhangxi MA Zhaopeng tang ying zhong Key Laboratory of Marine Ecology and Environmental Sciences Institute of OceanologyChinese Academy of SciencesQingdao 266071China Laboratory for Marine Ecology and Environmental Science Qingdao National Laboratory for Marine Science and TechnologyQingdao 266071China University of the Chinese Academy of Sciences Beijing 100049China

The accurate measurement of gene expression via quantitative real-time reverse transcription PCR（q RT-PCR）heavily relies on the choice of valid reference gene（s） for data normalization. Resting cyst is the dormant stage in the life cycle of dinoflagellate, which plays crucial roles in HAB-forming dinoflagellate ecology. However, only limited investigations have been conducted on the reference gene selection in dinoflagellates. Gap remained in our knowledge about appropriate HKGs for normalizing gene expression in different life stages, which laid obstacles for the application of q RT-PCR to the HAB-forming group. In this study, six candidate reference genes,18 S ribosomal RNA（18S）, glyceraldehyde-3-phosphate dehydrogenase（GAPDH）, α-tubulin（TUA）, β-tubulin（TUB）, actin（ACT） and cytochrome oxidase subunit 1（COX1）, were evaluated for their expression stability with q RT-PCR and three statistical algorithms（Ge Norm, Norm Finder, and Best Keeper） for the cosmopolitan, harmful algal bloom-forming dinoflagellate Akashiwo sanguinea. Expression patterns were observed across 18 biological samples, including cells at resting stages（resting cysts）, different growth stages, in darkness, exposed to abscisic acid（ABA） and exposed to temperature stress. The results indicated that TUA, 18 S and GAPDH were relatively stable across all tested scenarios. While the best-recommended reference genes differed across experimental groups, the pairs of ACT and TUA, 18 S and GAPDH were the most reliable for cells at different growth stages and darkness treatment. The combination of TUA and TUB was the best choice for normalization in resting cysts and in ABA treatment, respectively. The pair of ACT and COX1 was suitable for temperature treatments. This study was the first to investigate the stable internal reference genes in dinoflagellates at different stages of life cycle,particularly in resting cysts. Our results provided useful information for selection of reference genes in dinoflagellates

关键词： Akashiwo sanguinea dinoflagellate internal control qRT-PCR resting cyst reference gene

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The morphological and phylogenetic characterization for the dinoflagellate Margalefidinium fulvescens (=Cochlodinium fulvescens) isolated from the Jiaozhou Bay, China

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Acta Oceanologica Sinica 2018年第10期37卷 11-17页

作者： HU Zhangxi DENG Yunyan LI Yuhang tang ying zhong CAS Key Laboratory of Marine Ecology and Environmental Sciences Institute of OceanologyChinese Academy of SciencesQingdao 266071China Laboratory for Marine Ecology and Environmental Science Pilot National Laboratory for Marine Science and Technology(Qingdao)Qingdao 266237China Center for Ocean Mega-Science Chinese Academy of SciencesQingdao 266071China Department of Marine Organism Taxonomy and Phylogeny Institute of OceanologyChinese Academy of SciencesQingdao 266071China

The dinoflagellate genus Margalefidinium has been split from Cochlodinium as a new genus recently and Margalefidiniumfulvescens is one of the five Margalefidinium species. Margalefidiniumfulvescens is toxic and has been reported from the coastal waters of USA, Canada, Mexico, China, Japan, Indonesia, Korea, Pakistan and Spain. Here we provide the morphological and phylogenetic characterization for an isolate of it from the Jiaozhou Bay, Qingdao, China. Our results showed that the vegetative cells were subspherical to ellipsoidal, 34-60 pm in length, and 19-41 pm in width. Both single cell forms and colonies in chains of 2, 4, or 8 cells were observed in cultures, but chain forms with 2 or 4 cells were observed more often in the field samples. The cingulum was rather deep, encircling the cell approximately twice, but the sulcus was rather narrow, surrounding the cell about one turn. The nucleus was spherical and located at the central epicone. The chloroplasts were granular, brownish, and scattered peripherally. An orange pigmented body also appeared in the epicone. The apical groove appeared vase-like as previously described. Under epi-fluorescence microscopy, a pumpkin-like structure was clearly observed, in which cells were embedded. Cells were observed to exit from the structure, which led us to a hypothesis that the structure may provide cells a shelter to avoid predation or to respond to other stresses. The phylogenetic analyses based on partial LSU rDNA sequences indicated that M. fulvescens from the Jiaozhou Bay was grouped with M. fulvescens populations from other origins and closely related to the clade ofM. polykrikoides. Our morphological observations and phylogenetic analyses together confirmed the presence of M. fulvescens in China and our monitoring has also observed the species dominant in the dinoflagellate community of the Jiaozhou Bay in the early autumn of 2015, which alerted us to continually monitor this bloom-forming species in the region.

关键词： Margalefidiniumfulvescens morphology phylogeny pumpkin-like structure Jiaozhou Bay China

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基于BIM的智慧园区设计与全生命周期应用

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智能建筑与智慧城市 2024年第8期 140-142页

作者：唐中鹰上海德森建筑设计有限公司

文章分别从BIM技术在建筑设计的应用现状、智慧园区的发展需求、设计方法论、园区全生命周期管理等方面进行探讨,结合市场数据和实际案例阐述了BIM技术在智慧园区建设中的应用和效果。最后总结了BIM技术在应用中存在的技术难点及解决方... 详细信息

文章分别从BIM技术在建筑设计的应用现状、智慧园区的发展需求、设计方法论、园区全生命周期管理等方面进行探讨,结合市场数据和实际案例阐述了BIM技术在智慧园区建设中的应用和效果。最后总结了BIM技术在应用中存在的技术难点及解决方案,并对BIM技术全生命周期应用的节能减排、成本控制和运营效率提升效果进行了分析。

关键词： BIM 智慧园区全生命周期设计方法论市场数据

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当下高校民主党派基层组织建设问题及对策

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重庆文理学院学报（社会科学版） 2009年第4期28卷 18-20页

作者：唐忠英重庆文理学院经管学院重庆永川402160

高校民主党派基层组织建设面临的问题很多,文章选择部分突出问题进行研究,探讨解决途径,期望为高校民主党派基层组织的建设提供参考。

关键词：民主党派基层组织制度建设领导班子

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智能建筑电气施工管理及质量控制措施

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智能建筑与智慧城市 2024年第6期 142-144页

作者：唐中鹰上海德森建筑设计有限公司

文章从电气施工管理的重要性、智能建筑电气施工管理的基本理念和应用领域入手,探讨了智能建筑电气施工管理的基本功能、技术支持和质量控制措施,并结合实际案例进行了分析。最后,通过对市场数据、运营数据的分析,对智能建筑电气施工管... 详细信息

文章从电气施工管理的重要性、智能建筑电气施工管理的基本理念和应用领域入手,探讨了智能建筑电气施工管理的基本功能、技术支持和质量控制措施,并结合实际案例进行了分析。最后,通过对市场数据、运营数据的分析,对智能建筑电气施工管理的发展前景进行了展望。

关键词：智能建筑电气施工管理质量控制技术支持市场数据运营数据

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我国北部湾球形棕囊藻(Phaeocystis globosa)的表面形态和细胞超微结构的电镜观察

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海洋与湖沼 2019年第3期50卷 621-629页

作者：胡章喜邓蕴彦唐赢中中国科学院海洋生态与环境科学重点实验室(中国科学院海洋研究所) 青岛266071 青岛海洋科学与技术试点国家实验室海洋生态与环境科学功能实验室青岛266237 中国科学院海洋大科学研究中心青岛266071

本文首次对分离于我国广西北部湾的一株棕囊藻(Phaeocystis)纯培养进行了形态学研究。通过对其核糖体大亚基序列(LSU rDNA)进行了系统进化分析,并运用光学显微镜、扫描电镜和透射电镜等,对该藻的游动单细胞和囊体细胞表面结构和超微结... 详细信息

本文首次对分离于我国广西北部湾的一株棕囊藻(Phaeocystis)纯培养进行了形态学研究。通过对其核糖体大亚基序列(LSU rDNA)进行了系统进化分析,并运用光学显微镜、扫描电镜和透射电镜等,对该藻的游动单细胞和囊体细胞表面结构和超微结构进行了详细的观察。结果表明,通过分子序列和形态学特征确证了该纯培养为球形棕囊藻(Phaeocystis globosa)。通过分子系统进化分析,从北部湾获得的球形棕囊藻与同属的***和***的亲缘关系较近,但与***的亲缘关系相对较远。我们发现该藻至少有两种类型的细胞:具鞭毛游动细胞和无鞭毛囊体细胞,其大小分别为2.30-3.98μm和3.69-6.49μm。具鞭毛游动细胞表面具有1-2个位置不固定但形状较为规则的圆盘状凸起,细胞表面的鳞片大小无明显差别;囊体细胞均匀地分布在囊体上,囊体细胞表面光滑,有三个较短的附属物。具鞭毛游动细胞和囊体细胞有2个或4个叶绿体,具鞭毛游动细胞的细胞核位于细胞的中下部,而囊体细胞的细胞核则位于细胞的中上部。细胞分泌物在扫描电镜样品处理过程后形成多数与细胞相连的丝状物,且丝状物形成较为规则的五角星形结构,每个角与一根丝相连,有些丝状体上有"瘤状"结构。研究结果填补了我国在球形棕囊藻形态学和超微结构研究上的空白,为深入认识其生物学特征及其成囊机理提供基础。

关键词：球形棕囊藻表面形态超微结构系统进化

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Multi boron-doping effects in hard carbon toward enhanced sodium ion storage

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Journal of Energy Chemistry 2025年第1期100卷 730-738页

作者： Peng Zheng Wang Zhou ying Mo Biao Zheng Miaomiao Han Qin zhong Wenwen Yang Peng Gao Lezhi Yang Jilei Liu College of Materials Science and Engineering Hunan Joint International Laboratory of Advanced Materials and Technology for Clean Energy Hunan Province Key Laboratory for Advanced Carbon Materials and Applied Technology Hunan University Greater Bay Area Institute for Innovation Hunan University Changsha Research Institute of Mining & Metallurgy Co. LTD.

Hard carbon （HC） has been considered as promising anode material for sodium-ion batteries （SIBs）.The optimization of hard carbon’s microstructure and solid electrolyte interface （SEI） property are demonstrated effective in enhancing the Na+storage capability,however,a one-step regulation strategy to achieve simultaneous multi-scale structures optimization is highly ***,we have systematically investigated the effects of boron doping on hard carbon’s microstructure and interface chemistry.A variety of structure characterizations show that appropriate amount of boron doping can increase the size of closed pores via rearrangement of carbon layers with improved graphitization degree,which provides more Na+storage ***-situ Fourier transform infrared spectroscopy/electrochemical impedance spectroscopy （FTIR/EIS） and X-ray photoelectron spectroscopy （XPS） analysis demonstrate the presence of more BC3and less B–C–O structures that result in enhanced ion diffusion kinetics and the formation of inorganic rich and robust SEI,which leads to facilitated charge transfer and excellent rate *** a result,the hard carbon anode with optimized boron doping content exhibits enhanced rate and cycling *** general,this work unravels the critical role of boron doping in optimizing the pore structure,interface chemistry and diffusion kinetics of hard carbon,which enables rational design of sodium-ion battery anode with enhanced Na+storage performance.

关键词： X ray photoelectron spectroscopy

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四种海洋藻华甲藻与红色赤潮藻的化学互感作用(allelopathy)初步研究

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海洋科学 2017年第9期41卷 94-101页

作者：王欢胡章喜唐赢中中国科学院海洋生态与环境科学重点实验室中国科学院海洋研究所山东青岛266071 中国科学院大学北京100049 青岛海洋科学与技术国家实验室海洋生态与环境科学功能实验室山东青岛266071

化学互感作用或化感作用(allelopathy)在浮游植物的种群相互作用及动力学过程中起着重要的调节控制作用,已经成为有害藻华生态学研究中的重要领域。但在被研究的种类和化感作用的规律等方面还存在众多空白。本研究选取分离自中国和美国... 详细信息

化学互感作用或化感作用(allelopathy)在浮游植物的种群相互作用及动力学过程中起着重要的调节控制作用,已经成为有害藻华生态学研究中的重要领域。但在被研究的种类和化感作用的规律等方面还存在众多空白。本研究选取分离自中国和美国近海的重要藻华甲藻多环旋沟藻(Cochlodinium polykrikoides)、米氏凯伦藻(Karenia mikimotoi)、剧毒卡尔藻(Karlodinium veneficum)、短沟别什莱藻(Biecheleria brevisulcate),在相同的试验条件下(72 h共培养)观察其与另一种常见的藻华甲藻红色赤潮藻(Akashiwo sanguinea)的化感作用。实验结果显示化感效应随种类和特别是起始细胞密度比率而分别表现为抑制或促进共存种的生长。与起始细胞密度为500个/m L的红色赤潮藻共培养时,多环旋沟藻起始密度为250个/m L时促进红色赤潮藻生长,米氏凯伦藻苏澳株和短沟别什莱藻在细胞密度低于1 000个/m L时促进红色赤潮藻生长,同时起始密度为5 000~50 000个/m L的剧毒卡尔藻被红色赤潮藻促进生长。但是,米氏凯伦藻深圳湾株在细胞密度500个/m L以上时均抑制红色赤潮藻生长,多环旋沟藻在起始密度大于250个/m L时开始抑制红色赤潮藻生长,且抑制效应在起始密度2 500个/m L时接近100%。米氏凯伦藻的化感作用在不同株系间表现出显著差异。结果说明有害藻华甲藻之间的化感作用表现出复杂的形式,随着作用的种类和互相之间的密度比率变化而有互相促进、抑制或一方抑制(或促进)另一方等作用,这对进一步认识藻华甲藻的种群动力学具有重要意义。

关键词：化感作用红色赤潮藻多环旋沟藻剧毒卡尔藻米氏凯伦藻短沟别什莱藻

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