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Development of a real-time PCR method for Thalassiosira rotula rapid detection

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Acta Oceanologica Sinica 2007年第2期26卷 133-139页

作者： HE Shanying YU Zhigang mi tiezhu College of Chemistry and Chemical Engineering Ocean University of China Qingdao 266003 China College of Environmental Science and Engineering Ocean University of China Qingdao 266003 China

Gene specific primers and DNA probe were designed based on the sequence of 18S rDNA cloned from the red tide alga Thalassiosira rotula. A real-time fluorescent quantitative PCR （RFQ - PCR） method was developed for quantitative detection of T. rotula. The RFQ - PCR assay data showed that the results obtained with the RFQ - PCR quite good agreement with those with the light microscope （LM） counting method, which suggested that the RFQ - PCR could be a useful method for red tide alga detection.

关键词： red tide Thalassiosira rotula fluorescent quantitative PCR 18S rDNA

Effects of Ocean Acidification on Nitrogen Metabolism of Skeletonema costatum

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Journal of Ocean University of China 2024年第5期23卷 1359-1370页

作者： WANG Shuxing mi tiezhu ZHEN Yu ZHU Jianbin Key Laboratory of Marine Environment and Ecology Ministry of EducationCollege of Environmental Science and EngineeringOcean University of ChinaQingdao 266100China Laboratory for Marine Ecology and Environmental Science Qingdao Marine Science and Technology CenterQingdao 266237China Frontiers Science Center for Deep Ocean Multispheres and Earth System Ocean University of ChinaQingdao 266100China

Ocean acidification(OA),caused by the rising concentration of atmospheric CO_(2),leads to changes in the marine carbonate ***,in turn,affects the physiological processes of *** response to increased pCO_(2) levels,marine microalgae modulate their physiological responses to meet their energy and metabolic *** metabolism is a critical metabolic pathway,directly affecting the growth and reproductive capacity of marine *** the molecular mechanisms that regulate nitrogen metabolism in microalgae under OA conditions is therefore *** study aimed to investi-gate how OA affects the expression profiles of key genes in the nitrogen metabolic pathway of the marine diatom Skeletonema *** findings indicate that OA upregulates key genes involved in the nitrogen metabolic pathway,specifically those related to nitrate assimilation and glutamate ***,pCO_(2) has been identified as the predominant factor affecting the expression of these genes,with a more significant impact than pH variations in *** research not only advances our understanding of the adaptive mechanisms of *** in response to OA but also provides essential data for predicting the ecological consequences of OA on marine diatoms.

关键词： Skeletonema costatum ocean acidification nitrogen metabolism gene expression transcriptome

Relationship between bacteria and phytoplankton during the giant jellyfish Nemopilema nomurai bloom in an oligotrophic temperate marine ecosystem

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Acta Oceanologica Sinica 2016年第10期35卷 107-113页

作者： ZENG Yang HUANG Xuguang HUANG Bangqin mi tiezhu Key Laboratory of Coastal and Wetland Ecosystems of ministry of Education Xiamen University Fujian Provincial Key Laboratory of Coastal Ecology and Environmental Studies Xiamen University College of Chemistry and Chemical Engineering Ocean University of China

Bacterial abundance, phytoplankton community structure and environmental parameters were investigated to study the relationships between bacteria and phytoplankton during giant jellyfish Nemopilema nomurai blooms in the central Yellow Sea during 2013. N. nomurai appeared in June, increased in August, reached a peak and began to degrade in September 2013. Results showed that phosphate was possible a key nutrient for both phytoplankton and bacteria in June, but it changed to nitrate in August and September. Phytoplankton composition significantly changed that pico-phytoplankton relative biomass significantly increased, whereas other size phytoplankton significantly decreased during jellyfish bloom. In June, a significantly positive correlation was observed between chlorophyll a concentration and bacterial abundance（r=0.67, P0.001, n=34）.During jellyfish outbreak in August, there was no significant correlation between phytoplankton and bacteria（r=0.11, P0.05, n=25）, but the relationship（r=0.71, P0.001, n=31） was rebuilt with jellyfish degradation in September. In August, small size phytoplankton occupied the mixed layer in offshore stations, while bacteria almost distributed evenly in vertical. Chlorophyll a concentration significantly increased from（0.42±0.056） μg/L in June to（0.74±0.174） μg/L in August, while bacterial abundance just slightly increased. Additionally, the negative net community production indicated that community respiration was not entirely determined by the local primary productivity in August. These results indicated that jellyfish blooms potentially affect coupling of phytoplankton and bacteria in marine ecosystems.

关键词： jellyfish bloom Nemopilema nomurai phytoplankton pigments jellyfish DOM ecological consequence

Characterization of Bacterial Communities in Aerosols over Northern Chinese Marginal Seas and the Northwestern Pacific Ocean in Autumn

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Journal of Ocean University of China 2023年第1期22卷 136-150页

作者： ZHANG Bo ZHEN Yu mi tiezhu QI Jianhua YUAN Gang Key Laboratory of Marine Environment and Ecology Ministry of EducationOcean University of ChinaQingdao 266100China Laboratory for Marine Ecology and Environmental Science Pilot National Laboratory for Marine Science and Technology(Qingdao)Qingdao 266237China College of Environmental Science and Engineering Ocean University of ChinaQingdao 266100China

The characteristics of the bacterial community structure in aerosols of different particle sizes over northern Chinese marginal seas and the northwestern Pacific Ocean in autumn were analyzed using high-throughput sequencing and the quantitative real-time PCR method based on 16S rRNA *** with environmental factors and air mass sources,the bacterial abundance,community diversity,composition and structural characteristics in aerosols were studied,which might provide a scientific evaluation for a comprehensive understanding of the characteristics,long-distance transmission and ecological effects of aerosols from coastal waters to the distant ocean(northwestern Pacific Ocean).At the junction of the South Yellow Sea and the East China Sea,the bacterial abundance in the aerosols was substantially affected by transmission of continental air masses and was higher than those in other samples,and the bacterial abundance in coarse particle(>2.1μm)samples was significantly higher than that in fine particle(missions of continental air masses,and their bacterial abundance in coarse particle samples was significantly lower than that in fine particle *** differences in the richness and diversity of bacterial communities were observed among all samples(P0.05).Among the environmental factors examined,a significant negative correlation was observed between bacterial community richness and temperature(P<0.05),and the bacterial community diversity was significantly positively correlated with the concentration of K^(+)(P<0.05).Canonical correspondence analysis showed that temperature,NH_(4)^(+) concentration and SO_(4)^(2−) concentration exerted significant effects on bacterial community struc

关键词： bioaerosol marine environment community structure particle size

Molecular monitoring plankton community during jellyfish blooms in the water near the Liaoning Hongyan River Nuclear Power Plant

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Journal of Oceanology and Limnology 2023年第5期41卷 1781-1794页

作者： ming YUE tiezhu mi Ying LI Xianyu GONG Yu ZHEN Key Laboratory of Marine Environment and Ecology Ocean University of ChinaMinistry of EducationQingdao 266100China Laboratory for Marine Ecology and Environmental Science Pilot National Laboratory for Marine Science and Technology(Qingdao)Qingdao 266237China College of Environmental Science and Engineering Ocean University of ChinaQingdao 266100China

The Liaoning Hongyan River Nuclear Power Plant is located on the east coast of Liaodong Bay in the Bohai Sea,China,and the safety of the cooling water is often threatened by large-scale jellyfish ***,monitoring jellyfish growth,migration,and disappearance is essential to the safety of the nuclear power *** this study,high-throughput sequencing was used to analyze the community characteristics of mesoplankton in the sea area at the inlet and outlet from April to September *** the same time,the micro-individuals of jellyfish(including oosperm,planula,polyps,or ephyra of jellyfish)in the water were detected by real-time fluorescent quantitative PCR(qPCR).Both the maximum abundance of Aurelia coerulea and Nemopilema nomurai in July were detected by high-throughput sequencing and q *** three common taxa of plankton(Paracalanus,Calanoida,and Poaceae)were found in the waters near the inlet and outlet,and the beta diversity of plankton in the waters near the inlet and outlet was low in the same *** on the plankton community composition in the sea water near the inlet,it could be speculated that jellyfish outbreaks would have a significant impact on the plankton via the predation *** maximum mt-16S rDNA copy numbers of *** and *** in the samples from the waters of Liaodong Bay were detected in June 2019,and that of *** showed an increasing trend from north to south,with the jellyfish tending to move to the seawaters near the *** results indicate that the optimum monitoring period for jellyfish outbreaks in Liaodong Bay is from the end of May to the end of July.

关键词： jellyfish Aurelia coerulea Nemopilema nomurai molecular monitoring community

Asexual Reproduction of Aurelia coerulea Polyps in situ on Settling Plates in Jiaozhou Bay,China

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Journal of Ocean University of China 2024年第5期23卷 1371-1380页

作者： mi tiezhu ZHU Jianbin ZHEN Yu WANG Guoshan WANG Jianyan Key Laboratory of Marine Environment and Ecology Ministry of EducationCollege of Environmental Science and EngineeringOcean University of ChinaQingdao 266100China Laboratory for Marine Ecology and Environmental Science Qingdao Marine Science and Technology CenterQingdao 266237China Frontiers Science Center for Deep Ocean Multispheres and Earth System Ocean University of ChinaQingdao 266100China National Marine Hazard mitigation Service Ministry of Natural ResourcesBeijing 100194China Department of Life Sciences National Natural History Museum of ChinaBeijing 100050China

Jellyfish blooms are increasingly prevalent coastal phenomena,with Aurelia coerulea being a significant *** complex life history of jellyfish,featuring a sessile polyp stage,is a key factor in bloom formation,yet it is challenging to study in the *** study aimed to describe the asexual reproduction of *** polyps in Jiaozhou Bay,China,and to assess their ability to form sedentary colonies and their relationship with environmental *** settling plates,we observed polyps in situ and documented three asexual reproduction modes:budding,strobilation,and podocyst *** parameters including temperature,salinity,dissolved oxygen(DO),chlorophyll a(Chl a),and the biomass of phytoplankton and zooplankton were *** was the predominant reproductive method,featuring stolon budding in autumn and direct budding during the winter and spring *** took place in the colder months,releasing ephyrae between April and *** abundance was positively correlated with seawater temperature and negatively correlated with DO *** at 1.0 m depth showed higher survivorship and proliferation over 8 months,whereas those at 2.0 m depth experienced a steady decline and died within 4 *** study enhances our understanding of the life cycle and asexual reproduction strategies of *** polyps in the field,with implications for predicting and managing jellyfish *** findings underscore the importance of temperature and DO in polyp survival and suggest that initial polyp population proliferation is crucial for long-term survival.

关键词： jellyfish bloom polyps budding strobilation asexual reproduction

Influence of N, P, Fe Nutrients Availability on Nitrogen Metabolism-Relevant Genes Expression in Skeletonema marinoi

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Journal of Ocean University of China 2019年第1期18卷 239-252页

作者： JING Xiaoli mi tiezhu ZHEN Yu WANG Hualong YU Zhigang College of Marine Life Science Ocean University of China Function Laboratory for Marine Ecology and Environmental Science Qingdao National Laboratory for Marine Science and Technology Key Laboratory of Marine Environment and Ecology Ministry of Education College of Environmental Science and Engineering Ocean University of China Key Laboratory of Marine Chemical Theory and Technology Ministry of Education

Nitrogen(N), Phosphorus(P), and Iron(Fe) are essential elements for cellular structure and metabolism. In addition to dissolved inorganic nitrogen(DIN), phytoplankton is able to utilize dissolved organic nitrogen(DON). There is general consensus that both bacteria and higher plants nitrogen metabolism is affected by phosphate availability; this was also found to be true in coccolithophorid. Iron affects the structure and function of ecosystems through its effects on nitrogen metabolism. However, it is unclear how these nutrients affect Skeletonema marinoi's nitrogen metabolism. Here, using RT-qPCR, we investigate effects of N, P, and Fe on S. marinoi's nitrogen metabolism and nitrate reductase activity. These results illuminate that in S. marinoi, various nutrients have direct regulation on these genes expression at the molecular level. The varying degree of responses for these genes expression with differing N sources may act to increase the efficiency of nutrient capture when nitrate is limited. Suitable gene expression occurs at a N/P ratio of 16, which represents the atomic N/P ratio of phytoplankton cells and N/P concentrations in ocean; thus, nitrogen metabolism gene expression should be regulated by the existing N/P ratios in the phytoplankton's internal and external environment. Fe concentration has a direct and significant effect on nitrogen metabolism by regulating gene expression and nitrate reductase activity. Gene expression profiles identified in S. marinoi provide a foundation for understanding molecular mechanisms behind diatom nitrogen metabolism with changing N, P, and Fe nutrients allowing a basic understanding of how diatom growth is affected by nutrient utilization.

关键词： Skeletonema marinoi nitrogen metabolism relative gene expression RT-qPCR

Development of a Real-Time PCR Method (Taqman) for Rapid Identification and Quantification of Prorocentrum donghaiense

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Journal of Ocean University of China 2012年第3期11卷 366-374页

作者： YUAN Jian mi tiezhu ZHEN Yu YU Zhigang College of Marine Life Sciences Ocean University of ChinaQingdao 266003P.R.China College of Environmental Science and Engineering Ocean University of ChinaQingdao 266100P.R.China College of Chemistry and Chemical Engineering Ocean University of ChinaQingdao 266100P.R.China Key Laboratory of Marine Environment and Ecology Ministry of EducationOcean University of ChinaQingdao 266100P.R.China Key Laboratory of Marine Chemistry Theory and Technology Ministry of EducationOcean University of ChinaQingdao 266100P.R.China

Prorocentrum donghaiense is a dinoflagellate that is widely distributed in the East China Sea and has become increasingly involved in Harmful Algal Blooms (HABs). Therefore, it is necessary to study this dinoflagellate to monitor HABs. In this study, 13 pairs of primers specific to P. donghaiense (within its internal transcribed spacer (ITS) regions) were designed for SYBR Green I real-time PCR. As the SYBR Green I real-time PCR could not identify P. donghaiense in a specific manner, a Taqman real-time PCR method was developed by designing a set of specific primers and a Taqman probe. A 10-fold serial dilution of recombinant plasmid containing ITS regions of P. donghaiense was prepared as standard samples and the standard curve was established. Additionally, we quantified the genomic DNA in P. donghaiense cells and utilized this DNA to prepare another 10-fold serial dilution of standard sample and accordingly set up the standard curve. The mathematic correlation between the cell number and its corresponding plasmid copy number was also established. In order to test the efficiency of the real-time PCR method, laboratory samples and P. donghaiense HAB field samples were employed for identification and quantitative analysis. As to laboratory samples, as few as 102 cells of P. donghaiense could be quantified precisely utilizing both centrifugation and filtration techniques. The quantification results from field samples by real-time PCR were highly similar to those by light microscopy. In conclusion, the real-time PCR could be applied to identify and quantify P. donghaiense in HABs.

关键词： Prorocentrum donghaiense Harmful Algal Blooms (HABs) internal transcribed spacer (ITS) recombinant plasmid real-time PCR

microbial Community Composition and Function in Sediments from the Pearl River Mouth Basin

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Journal of Ocean University of China 2020年第4期19卷 941-953页

作者： CHEN Ye mi tiezhu LIU Yueteng LI Siqi ZHEN Yu College of Marine Life Sciences Ocean University of ChinaQingdao 266003China Laboratory for Marine Ecology and Environmental Science Pilot National Laboratory for Marine Science and Technology(Qingdao)Qingdao 266071China Key Laboratory of Marine Environment and Ecology Ministry of EducationQingdao 266100China College of Environmental Science and Engineering Ocean University of ChinaQingdao 266100China

This study was conducted to characterize the diversity and function of microbial communities in marine sediments of the Pearl River Mouth Basin(PRMB)in the South China *** results showed that the bacterial and archaeal communities varied greatly with *** in bacterial communities and Nitrososphaeria and Woesearchaeota in archaeal communities were dominant in the shallow sediments(1-40 cm),while Chloroflexi in bacterial communities and Bathyarchaeia in archaeal communities were dominant in the deep sediments(50-200 cm).Regarding ecological functions based on the metatranscriptomic data,genes involved in various pathways of nitrogen metabolism and sulfur metabolism were observed in the tested sediment *** analysis revealed that Proteobacteria contribute the most to nearly all genes involved in nitrogen and sulfur ***,Thaumarchaeota contribute the most to certain genes involved in nitrification,denitrification and assimilatory sulfate reduction *** most abundant bacterial genus,Candidatus Scalindua,is crucial for nitrification,dissimilatory nitrate reduction,denitrification and assimilatory sulfate reduction pathways.

关键词： microbial community and function high-throughput sequencing Pearl River Mouth Basin(PRMB)