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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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Blooms of Prorocentrum donghaiense reduced the species diversity of dinoflagellate community

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Acta Oceanologica Sinica 2020年第4期39卷 110-119页

作者： Huan Wang Zhangxi Hu Zhaoyang Chai yunyan deng Zifeng Zhan Ying Zhong Tang 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 University of Chinese Academy of Sciences Beijing 100049China Center for Ocean Mega-Science Chinese Academy of SciencesQingdao 266071China Department of Marine Organism Taxonomy and Phylogeny Institute of OceanologyChinese Academy of SciencesQingdao 266071China

Most of reported harmful algal blooms(HABs)of microalgae(75%)have been caused by *** on the negative effects of HABs have generally focused on animals,valuable organisms in particular,and environmental factors such as dissolved oxygen and nutrients,but relatively fewer on community level,particularly that using metagenomic *** this study,we reported an investigation on the effects of a HAB caused by the dinoflagellate Prorocentrum donghaiense on the species diversity and community structure of the dinoflagellate sub-community via a pyrosequencing approach for the samples taken before,during,and after the bloom season of *** in the East China *** sequenced partial 28S rRNA gene of dinoflagellates for the field samples and evaluated the species richness and diversity indices of the dinoflagellate community,as a sub-community of the total *** obtained 800185 valid sequences(categorized into 560 operational taxonomic units,OTUs)of dinoflagellates from 50 samples and found that the biodiversity of dinoflagellate community was significantly reduced during the blooming period in comparison to that in pre-and after-blooming periods,as reflected in the four diversity indices:the species richness expressed as the number of OTUs,Chao1 index,Shannon index(evenness),and Gini-Simpson *** four indices were all found to be negatively correlated to the cell density of the bloom species *** analyses also revealed that the *** cell abundance was correlated negatively with NO3--N,and NO2--N,but positively with total nitrogen(TN)and total phosphorus(TP).Principal coordinates analysis(PCoA)showed that the community structure of dinoflagellates was markedly different among the different sampling periods,while the redundancy analysis(RDA)revealed *** abundance,salinity,NO3--N,and SiO32-were the most four significant factors shaping the dinoflagellate community *** results toget

关键词： Prorocentrum donghaiense dinoflagellate community diversity pyrosequencing East China Sea

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Individual differences determine the morphogenesis pattern and reproductive uniqueness of the invasive green macroalga,implications for its habitat adaptation

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Journal of Oceanology and Limnology 2024年第6期42卷 2013-2023页

作者： Bingxin HUANG Liming RAO yunyan deng Boxi LI Zhiying WANG Jie CHEN Lichun WANG Yang DAI Lanping DING College of Life Sciences Tianjin Normal UniversityTianjin 300387China Institute of Oceanology Chinese Academy of SciencesQingdao 266071China Tianjin Key Laboratory of Animal and Plant Resistance Tianjin 300387China

The marine green algae genus Chaetomorpha is a common source of“green tide”and is widespread on coasts around the *** this study,based on invasive Chaetomorpha valida collected from the Shandong Peninsula,the morphogenesis and reproductive characteristics of two strains that are morphologically different from each other,were observed using experimental biology *** main results are as follows:(1)significant difference in the size of reproductive cells produced by Strains 1 and 2;(2)gametes produced by Strain 2 are isogamous and same gametangial during the binding process of gametes,whereas those of Strain 1 are isogamous but hetero-cystic;(3)progeny from Strain 1 has rhizoidal holdfast,whereas that of Strain 2 has discoid holdfast;(4)gametophytic“branching”was found in Strain *** results could validate the high phenotypic plasticity of macroalgae and offered an interpretation of habitat ***,this study innovatively provided fundamental research on the selection of macroalgal traits and explored competitive strategies for the dominant survival of macroalgae from a new perspective.

关键词： Chaetomorpha valida individual difference morphogenesis pattern reproductive uniqueness

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Reviving and characterizing three species of dinoflagellate cysts dormant for about 70 years in the East China Sea:Biecheleria brevisulcata,Biecheleriopsis adriatica,and Scrippsiella donghaienis

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Journal of Oceanology and Limnology 2022年第6期40卷 2292-2311页

作者： Zhangxi HU Xiaoying SONG Jinxiu WANG Zhe TAO Yuanyuan SUN Yuhang LI Yuyang LIU yunyan deng Lixia SHANG Zhaoyang CHAI Yingzhong TANG College of Fisheries Guangdong Ocean UniversityZhanjiang 524088China CAS Key Laboratory of Marine Ecology and Environmental Sciences Institute of OceanologyChinese Academy of SciencesQingdao 266071China Laboratory of Marine Ecology and Environmental Science Pilot National Laboratory for Marine Science and Technology(Qingdao)Qingdao 266237China CAS Key Laboratory of Experimental Marine Biology Institute of OceanologyChinese Academy of SciencesQingdao 266071China Laboratory of Marine Organism Taxonomy and Phylogeny Institute of OceanologyChinese Academy of SciencesQingdao 266071China Center for Ocean Mega-Science Chinese Academy of SciencesQingdao 266071China University of Chinese Academy of Sciences Beijing 100049China

Many marine dinoflagellates can form resting cysts as a part of their life cycle,and the cysts could be buried in sediment and remained viable for as long as over 150 ***,only a very limited number of cyst species have been revived from long-buried sediments and investigated in regard to a possible shift in the intra-specific genetic structure of a species detected from the historical record at a particular ***,we report a successful germination of three species of resting cysts that were sampled from the depth dated back to 1941±18 AD from a 44-cm sediment core from the East China *** isolates were established from germination of single cyst isolation or multi-cyst *** rRNA gene or ITS sequences of these strains were obtained,then they were identified to be Biecheleria brevisulcata(five strains),Biecheleriopsis adriatica(one strain),and Scrippsiella donghaienis(one strain)in terms of morphology and rRNA gene *** brevisulcata strain 1,*** strain 21,and *** strain 23 were examined in detail with light microscope(LM)and scanning electron microscope(SEM),and analyzed with high performance liquid chromatography(HPLC)for their pigment compositions,and genetic *** also confirmed the presence of a resting cyst of *** in the field for the first *** LSU rRNA gene-based genetic distances of *** from that obtained from water sample,single-cell PCR sequencing for the cysts isolated from the surface sediment of the same sea area and that reported from other regions during the recent years,and ITS-based genetic distances of *** from that obtained from cysts isolated from the surface sediment of the same location and that reported from other regions during the recent years indicated that the intra-specific genetic structure of each species in the sampling area may have shifted during the last 70 *** work confirms that ***,***,and ***

关键词： core sediment dinoflagellate resting cyst germination Biecheleria brevisulcata Biecheleriopsis adriatica Scrippsiella donghaienis

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叙事医学在儿科学“医患沟通”课程中的应用探讨

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中国医学教育技术 2020年第6期34卷 791-794,798页

作者：孙凤杰林天云李娟邓云燕广州医科大学附属第五医院新生儿科广州510700

目的探讨叙事医学应用于儿科学住院医师规范化培训过程中医患沟通的作用。方法选取广州医科大学2017—2019级儿科学专业住院医师规范化培训学员80人,采用随机数字表以及随机数余数分组法将学员分为对照组和实验组,每组各40人;对照组采... 详细信息

目的探讨叙事医学应用于儿科学住院医师规范化培训过程中医患沟通的作用。方法选取广州医科大学2017—2019级儿科学专业住院医师规范化培训学员80人,采用随机数字表以及随机数余数分组法将学员分为对照组和实验组,每组各40人;对照组采用课堂授课方式学习“医患沟通”课程,实验组采用叙事医学方式分析并演绎医患关系的经典案例,比较两组学员的学习提升程度、成绩考核分数与知识掌握程度、课堂教学效果及教学方法的满意度。结果实验组学员的学习提升程度、成绩考核分数与知识掌握程度、课堂教学效果均高于对照组(P<0.05);实验组学员对教学方法的满意度高(P<0.05)。结论叙事医学对优化“医患沟通”课堂气氛、提高医患沟通技巧具有积极作用,为保证医疗服务质量及提高医疗技术水平提供支持。

关键词：叙事医学儿科学住院医师规范化培训

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Preliminary Study on the Life History of Chaetomorpha valida（Cladophoraceae,Chlorophyta） in laboratory culture

Preliminary Study on the Life History of Chaetomorpha valida...

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庆祝中国藻类学会成立30周年暨第十五次学术讨论会

作者： yunyan deng~(1,2) Xiaorong Tang~3 Lanping Ding~(1**) and Shaoxing Lian~1 (1.Institute of Oceanology,Chinese Academy of Sciences,Qingdao 266071,China 2.Graduate University of Chinese Academy of Sciences,Beijing 100049,China 3.Marine Genetics and Breeding Laboratory,Ocean University of China,Qingdao 266003,China)

＜正＞Chaetomorpha valida Kützing,as a fouling green alga,was found in aquaculture ponds in coast of northern China recent *** spring and summer,mass occurrence of the macroalga causes great damage to *** on the unialgal culture and microscope observation,development stages of life cycle of the species were studied. Its life history is typical alternation of generations between isomorphic,unisexual haploid gametophytes and diploid *** and female gametes are released from gametangia of gametophytes to fuse into zygotes and settled,then developed into new *** reproduction was presented by quadriflagellate *** germination pattern of both zygote and zoospore were a bipolar erect type as observed in some other species of *** results also contribute to increase the understanding about seasonal abundances of the macroalgae.

关键词： Chaetomorpha valida life history unialgal culture alternation of generations

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A new record species of epiphytic marine green algae,Entocladia leptochaete（Chaetophoraceae,Chlorophyta）with its primary experimental biology from China

A new record species of epiphytic marine green algae,Entocla...

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庆祝中国藻类学会成立30周年暨第十五次学术讨论会

＜正＞Entocladia leptochaete,a new record species in Chaetophoraceae（Chlorophyta）, was observed during our recent laboratory culture of marine benthic green alga, Chaetomorpha *** mainly morphological characteristics of the species are as follows:Thallus is composed of microscopic,prostrate,uniseriate branched filaments. Elongated filaments with some short lateral branches consolidate centrally into *** branches in the external frond are rectangular to slightly swollen and sometimes irregular or distorted at the *** at external frond is rounded or irregular in shape,7-14（-17）μm diameter,14-33μm long,3-5 times as long as *** of cells have 2-4（-6） pyrenoids, occasionally single with diameter of pyrenoids ranging from 2.5μm to 5μ*** parietal chloroplasts are slightly lobed. The first report of this species in China will bridge its distributional range from Europe to *** morphologies,taxonomy and distribution were introduced and discussed in *** culture experiments at different temperatures and light intensities were also carried *** results suggested that the species could adapt well to extensively aquicolous environment.

关键词： China Entocladia leptochaete morphology and taxonomy new record experimental biology

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耳穴贴压法治疗妊娠恶阻临床疗效的Meta分析

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中西医结合护理 2022年第6期8卷 8-12页

作者：田恬谢薇邓云燕李忠琴张杨赵苏贵州中医药大学第一附属医院产科优生研究中心贵州贵阳550001 贵州中医药大学第一附属医院护理部贵州贵阳550001

目的评价耳穴贴压法治疗妊娠恶阻的临床效果。方法通过计算机检索中国知网(CNKI)、维普数据库(VIP)、万方数据库(Wanfang Database)、PubMed及Cochrane library,收集耳穴贴压法治疗妊娠恶阻的随机对照试验,按照Jadad量表中随机隐藏的质... 详细信息

目的评价耳穴贴压法治疗妊娠恶阻的临床效果。方法通过计算机检索中国知网(CNKI)、维普数据库(VIP)、万方数据库(Wanfang Database)、PubMed及Cochrane library,收集耳穴贴压法治疗妊娠恶阻的随机对照试验,按照Jadad量表中随机隐藏的质量标准来评价所纳入研究的质量。采用Rev Man 5.3软件进行Meta分析。结果共纳入5篇研究,包括359例妊娠恶阻患者。Meta分析结果显示:①耳穴贴压组在治愈率上反应的疗效优于基础治疗组[MD=2.42,95%CI(2.14,5.45),Z=5.16,P<0.00001]。②耳穴贴压组在总有效率上反应的疗效优于基础治疗组[MD=5.01,95%CI(2.495,-10.07),Z=4.52,P<0.00001]。结论耳穴贴压法治疗妊娠恶阻优于单纯基础治疗,但耳穴贴压法的起效机制尚未明确,需有待研究。

关键词：耳穴贴压耳穴埋豆耳穴埋籽妊娠恶阻 Meta分析

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Marxism and the Future of Western Aesthetics——The Third Sino-British Bilateral Forum on Marxist Aesthetics

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外国文学研究 2013年第5期35卷 171-174页

作者： Zhang yunyan deng Wenyu School of Humanities Shanghai Jiao Tong University Shanghai Finance and Economics University

An event themed the Third Sino-British Bilateral Forum on Marxist Aesthetics,jointly organized by School of Humanities in Shanghai Jiao Tong University and the School of Arts,Language and Cultures,University of Manchester,was held at Shanghai Jiao Tong University from April 6th to 8th,*** more than 17 foreign universities and about 80 scholars from over 50 domestic universities,research institutes and media attended the meeting.

关键词：外国文学文学创作文学评论文学作品

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