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Journal of Environmental Sciences 2015年第4期27卷 241-251页

作者： Min Wei Zhisheng yu Hongxun Zhang College of Resources and Environment University of Chinese Academy of Sciences

Microbial diversity and abundance in bioaerosols of a coal mine were analyzed based on 454 pyrosequencing and real-time polymerase chain reaction（PCR）. A total of 37,191 high quality sequences were obtained and could be classified into 531, 1730 and 448 operational taxonomic units respectively for archaea, bacteria and fungi at 97% sequence similarity. The Shannon diversity index for archaea, bacteria and fungi was respectively 4.71, 6.29 and 3.86, indicating a high diversity in coal mine bioaerosols. Crenarchaeota, Proteobacteria and Ascomycota were the dominant phyla for archaea, bacteria and fungi, respectively. The concentrations of total archaea, bacteria and fungi were 1.44 × 10^8, 1.02 × 10^8 and 9.60 × 10^4cells/m^3, respectively.Methanotrophs observed in bioaerosols suggested possible methane oxidation in the coal mine. The identified potential pathogens to coal miners, such as Acinetobacter schindleri,Aeromonas cavernicola, Alternaria alternata, Aspergillus penicillioides, Cladosporium cladosporioides,and Penicillium brevicompactum were also observed. This was the first investigation of microbial diversity and abundance in coal mine bioaerosols. The investigation of microbial communities would be favorable in promoting the progress of methane control based on microbial technique and concern on coal miners＇ health.

关键词： Microbial community Bioaerosols Methanotrophs Coal mine

Unravelling how biochar and dung amendments determine the functional structure and community assembly related to methane metabolisms in grassland soils

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Biochar 2022年第1期4卷 377-390页

作者： Qingzhou Zhao Yanfen Wang Zhihong Xu Juanli yun Zhisheng yu College of Resources and Environment University of Chinese Academy of SciencesBeijing 100049People’s Republic of China College of Urban and Environmental Science Peking UniversityBeijing 100871People’s Republic of China School of Environment and Science Griffith UniversityNathanQLD 4111Australia Center of Planetary Health and Food Security Griffith Univer-sityNathanQLD 4111Australia Yanshan Earth Critical Zone and Surface Flux Observation Research Station Chinese Academy of SciencesBeijing 100049People’s Republic of China State Key Laboratory of Microbial Resources Institute of MicrobiologyChinese Academy of SciencesBeijing 100101People’s Republic of China RCEES-IMCAS-UCAS Joint-Lab of Microbial Technology for Environmental Science Beijing 100085People’s Republic of China

Biochar and dung amendments have been extensively employed in soil remediation and fertilization of grasslands,which are the largest terrestrial sinks for methane.However,how these exogenous amendments regulate methane metabolisms at the molecular and community levels remains elusive.In this study,we investigated the functional genes and community assemblies of methanogens and methanotrophs using Geochip 5.0 and high-throughput sequencing to reveal the impacts of biochar and dung on soil methanogenesis and methane oxidation.The interac-tions between methane metabolic genes and other biogeochemical genes were also examined.According to Geo-chip microarrays,methanogenic gene mcrA decreased and increased with dung or biochar amendment,respectively;The methanotrophic gene pmoA showed a reverse but not significant tendency.Undominated processes contributed 65.51%to replace homogeneous selections as primary driving forces of methanogen assembly after dung amend-ment;the contribution of dispersal limitation increased to 46.13%in methanotroph assembly after biochar amend-ment.The diversity and association of co-occurrence networks for carbon-nitrogen cycling genes decreased after exogenous amendments.These results indicated that biochar and dung amendments prominently regulated the functional genes and community assembly involved in methane metabolisms.The co-existence patterns of methane metabolic genes and other related geochemical genes were also shaped by these amendments.This study provides the scientific reference for the development of grassland management in the context of global warming.

关键词： Biochar amendment Dung amendment Methane metabolism Nitrogen cycling Community assembly Geochip 5.0

Validation of Bacteroidales-based microbial source tracking markers for pig fecal pollution and their application in two rivers of North China

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Frontiers of Environmental Science & Engineering 2020年第4期14卷 139-146页

作者： Youfen Xu Zong Li Ruyin Liu Hongxia Liang Zhisheng yu Hongxun Zhang College of Resources and Environment University of Chinese Academy of SciencesBeijing 100049China

In China,pig feces is the predominant source of excrement produced b y animal husbandry.Improper use or direct discharge of pig feces can result in contamination of natural water systems.Microbial source tracking(MST)technology can identify the sources of fecal pollution in environmental water,and contribute to the management of pig fecal pollution by local environmental protection agencies.However,the accuracy of such assays can be context-dependent,and they have not been comprehensively evaluated under Chinese conditions.We aimed to compare the performance of five previously reported nig-specific MST assays(PF,Pig-Bac 1 STBR,Pig-Bac2 SYBR,Pig-1-Bac TaqMan,and Pig-2-Bac TaqMan,which are based on Bacteroidales 16S rRNA gene markers)and apply them in two rivers of North China.We collected a total of 173 fecal samples from pigs,cows,goats,chickens,humans,and horses across China.The PF assay optimized in this study showed outstanding qualitative performance and achieved 100%specificity and sensitivity.However,the two SYBR green qPCR assays(Pig-Bac1 SYBR and Pig-Bac2 SYBR)cross-reacted with most non-pig fecal samples.In contrast,both the Pig-l-Bac TaqMan and Pig-2-Bacr TaqMan assays gave 100%specificity and sensitivity.Of these,the Pig-2-Bac TaqMan assay showed higher reproducibility.Our results regarding the specificity of these pig-specific MST assays differ from those reported in Thailand,Japan,and America.Using the PF and Pig-2-Bac TaqMan assays,a field test comparing the levels of pig fecal pollution in rivers near a pig farm before and after comprehensive environmental pollution governance indicated that pig fecal pollution was effectively controlled at this location.

关键词： Microbial source tracking Pig fecal pollution 16S rRNA genem arkers Pig-specific Bacteroidales

Inhibitor tolerance and bioethanol fermentability of levoglucosan-utilizing Escherichia coli were enhanced by overexpression of stress-responsive gene ycfR:The proteomics-guided metabolic engineering

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Synthetic and Systems Biotechnology 2021年第4期6卷 384-395页

作者： Dongdong Chang Zia Ul Islam Junfang Zheng Jie Zhao Xiaoyong Cui Zhisheng yu College of Resources and Environment University of Chinese Academy of SciencesBeijing100049PR China Department of Chemistry Mississippi State UniversityMississippi StateMS 39762USA Department of Biochemistry and Molecular Biology School of Basic Medical SciencesCapital Medical UniversityBeijing100069PR China Yanshan Earth Critical Zone and Surface Fluxes Research Station Chinese Academy of SciencesNo.380 Huaibei TownHuairou DistrictBeijing101408PR China RCEES-IMCAS-UCAS Joint-Lab of Microbial Technology for Environmental Science Beijing100085PR China

Pretreatment of lignocellulosic biomass is crucial for the release of biofermentable sugars for biofuels production,which could greatly alleviate the burgeoning environment and energy crisis caused by the massive usage of traditional fossil fuels.Pyrolysis is a cost-saving pretreatment process that can readily decompose biomass into levoglucosan,a promising anhydrosugar;however,many undesired toxic compounds inhibitory to downstream microbial fermentation are also generated during the pyrolysis,immensely impeding the bioconversion of levoglucosan-containing pyrolysate.Here,we took the first insight into the proteomic responses of a levoglucosan-utilizing and ethanol-producing Escherichia coli to three representative biomass-derived inhibitors,identifying large amounts of differentially expressed proteins(DEPs)that could guide the downstream metabolic engineering for the development of inhibitor-resistant strains.Fifteen up-and eight down-regulated DEPs were further identified as the biomarker stress-responsive proteins candidate for cellular tolerance to multiple inhibitors.Among these biomarker proteins,YcfR exhibiting the highest expression fold-change level was chosen as the target of overexpression to validate proteomics results and develop robust strains with enhanced inhibitor tolerance and fermentation performance.Finally,based on four plasmid-borne genes encoding the levoglucosan kinase,pyruvate decarboxylase,alcohol dehydrogenase,and protein YcfR,a new recombinant strain E.coli LGE-ycfR was successfully created,showing much higher acetic acid-,furfural-,and phenol-tolerance levels compared to the control without overexpression of ycfR.The specific growth rate,final cell density,ethanol concentration,ethanol productivity,and levoglucosan consumption rate of the recombinant were also remarkably improved.From the proteomics-guided metabolic engineering and phenotypic observations,we for the first time corroborated that YcfR is a stress-induced protein responsive to multiple biomass-derived inhibitors,and al

关键词： Lignocellulosic biomass Levoglucosan Inhibitor Proteomics Gene overexpression Bioethanol

Enhanced straw fermentation process based on microbial electrolysis cell coupled anaerobic digestion

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Chinese Journal of Chemical Engineering 2022年第4期35卷 239-245页

作者： Xinyu Yan Bobo Wang Hongxia Liang Jie Yang Jie Zhao Fabrice Ndayisenga Hongxun Zhang Zhisheng yu Zhi Qian College of Resources and Environment University of Chinese Academy of SciencesBeijing 100049China Research Center for Eco-Environmental Sciences-Institute of Microbiology Chinese Academy of Sciences-University of Chinese Academy of Sciences(RCEES-IMCAS-UCAS)Joint-Lab of Microbial Technology for Environmental ScienceBeijing 100085China Received:2020-11-12 Revised:2021-05-08 Online:2022-04-28 Published:2022-06-18

The low quality and yield of methane severely hinder the industrial application of straw biogas fermentation, and no effective solution has been found so far. In this study, a novel method was developed when a microbial electrolysis cell(MEC) was coupled with normal anaerobic fermentation to enhance methane yield and purity. The fermentation process achieved a methane purity of more than 85%, which is considerably higher than that of previously published reports. With microbial stimulation and an electric current, the degradation of fibers has been greatly enhanced. The MEC system substantially improved the yield and purity of biogas, bringing a new path to the synthesis of methane by carbon dioxide and hydrogen ions in solution under electron irradiation. Electrochemical index analysis showed extra methane synthesis, due to the external circuit electron transfer. The results of the gas chromatography and solid degradation rate showed that the carbon source of extra methane was CO_(2) produced during normal fermentation and additional volatile solid degradation. These results show that the MEC considerably enhanced the quality and yield of methane in the straw fermentation process, providing insights into normal anaerobic fermentation.

关键词： Microbial electrolysis cell(MEC) Methane Straw Fermentation Bioenergy

The transcriptional activator Klf5 recruits p300-mediated H3K27ac for maintaining trophoblast stem cell pluripotency

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Journal of Molecular Cell Biology 2023年第7期15卷 31-44页

作者： Chengli Dou Linhui Wu Jingjing Zhang Hainan He Tian Xu Zhisheng yu Peng Su Xia Zhang Junling Wang Yi-Liang Miao Jilong Zhou Institute of Stem Cell and Regenerative Biology College of Animal Science and Veterinary MedicineHuazhong Agricultural UniversityWuhan 430070China Key Laboratory of Agricultural Animal Genetics Breeding and Reproduction(Huazhong Agricultural University)Ministry of EducationWuhan 430070China Department of Reproductive Medicine Huangshi Central HospitalAffiliated Hospital of Hubei PolytechnicEdong Healthcare GroupHuangshi 435000China Hubei Hongshan Laboratory Wuhan 430070China

The effective proliferation and differentiation of trophoblast stem cells(TSCs)is indispensable for the development of the placenta,which is the key to maintaining normal fetal growth during pregnancy.Kruppel-like factor 5(Klf5)is implicated in the activation of pluripotency gene expression in embryonic stem cells(ESCs),yet its function in TSCs is poorly understood.Here,we showed that Klf5 knockdown resulted in the downregulation of core TSC-specific genes,consequently causing rapid differentiation of TSCs.Consistently,Klf5-depleted embryos lost the ability to establish TSCs in vitro.At the molecular level,Klf5 preferentially occupied the proximal promoter regions and maintained an open chromatin architecture of key TSC-specific genes.Deprivation of Klf5 impaired the enrichment of p300,a major histone acetyl transferase of H3 lysine 27 acetylation(H3K27ac),and further reduced the occupancy of H3K27ac at promoter regions,leading to decreased transcriptional activity of TSC pluripotency genes.Thus,our findings highlight a novel mechanism of Klf5 in regulating the self-renewal and differentiation of TSCs and provide a reference for understanding placental development and improving pregnancy rates.

关键词： Klf5 trophoblast stem cells self-renewal chromatin accessibility p300 H3K27ac

Soft-rigid coupling grippers:Collaboration strategies and integrated fabrication methods

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Science China(Technological Sciences) 2023年第11期66卷 3051-3069页

作者： CHEN Han ZHU JiaQi CAO yu XIA zhisheng CHAI ZhiPing DING Han WU ZhiGang State Key Laboratory of Digital Manufacturing Equipment and Technology School of Mechanical Science and EngineeringHuazhong University of Science and TechnologyWuhan 430074China Department of Mechanical and Automation Engineering The Chinese University of Hong KongHong Kong SAR 999077China

Continuously increasing applications of robot technologies in unstructured environments put higher requirements on the robotic grippers'performance,such as interaction capability,output force range,and controllability.However,currently,it is hard for either rigid or soft grippers to meet these requirements,as single soft or rigid structures alone are difficult to effectively overcome/alleviate their inherent defects,e.g.,low compliance of rigid structures and low output force of soft structures.To deal with these difficulties,soft-rigid coupling grippers,or hybrid grippers are proposed.Technically,the soft-rigid coupling is a promising design that combines soft and rigid structures,in order to exploit their respective advantages,such as the strength of rigid structures and compliance of soft structures,in the same set of the gripper system.For the first time,herein,this paper systematically discusses the collaboration strategies of the existing hybrid robotic grippers,by classifying them as Rigid-activesoft-passive,Rigid-passive-soft-active,and Rigid-active-soft-active.At the same time,we introduce the integrated fabrication methods of hybrid grippers,through which the soft and rigid structures with great stiffness and property differences can be coupled together to construct a stable system.Also,possible performance improvements on soft-rigid coupling design for gripper systems are discussed.

关键词： grasping grippers hybrid design soft-rigid coupling soft-rigid collaboration

Molecular analysis of long-term biofilm formation on PVC and cast iron surfaces in drinking water distribution system

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Journal of Environmental Sciences 2014年第4期26卷 865-874页

作者： Ruyin Liu Junge Zhu Zhisheng yu DevRaj Joshi Hongxun Zhang Wenfang Lin Min Yang College of Environmental and Resource Sciences University of Chinese Academy of Sciences Institute of Microbiology Chinese Academy of Sciences Research Center for Eco-Environmental Sciences Chinese Academy of Sciences

To understand the impacts of different plumbing materials on long-term biofilm formation in water supply system, we analyzed microbial community compositions in the bulk water and biofilms on faucets with two different materials-polyvinyl chloride （PVC） and cast iron, which have been frequently used for more thanlO years. Pyrosequencing was employed to describe both bacterial and eukaryotic microbial compositions. Bacterial communities in the bulk water and biofilm samples were significantly different from each other. Specific bacterial populations colonized on the surface of different materials. Hyphomicrobia and corrosion associated bacteria, such as Acidithiobacillus spp., Aquabacterium spp., Limnobacter thiooxidans, and Thiocapsa spp., were the most dominant bacteria identified in the PVC and cast iron biofilms, respectively, suggesting that bacterial colonization on the material surfaces was selective. Mycobacteria and Legionella spp. were common potential pathogenic bacteria occurred in the biofilm samples, but their abundance was different in the two biofilm bacterial communities. In contrast, the biofilm samples showed more similar eukaryotic communities than the bulk water. Notably, potential pathogenic fungi, i.e., Aspergillus spp. and Candida parapsilosis, occurred in similar abundance in both biofilms. These results indicated that microbial community, especially bacterial composition was remarkably affected by the different pipe materials （PVC and cast iron）.

关键词： drinking waterbiofilm surface material pyrosequencing microbial community

Analysis of bacterial community in bulking sludge using culture-dependent and -independent approaches

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Journal of Environmental Sciences 2011年第11期23卷 1880-1887页

作者： Decai Jin Ping Wang Zhihui Bai Xinxin Wang Hong Peng Rong Qi Zhisheng yu Guoqiang Zhuang Research Centre for Eco-Environmental Sciences Chinese Academy of SciencesBeijing 100085China Graduate University of Chinese Academy of Sciences Beijing 100049China School of Chemical Engineering The University of QueenslandBrisbaneQueensland 4072Australia

The bacterial community of a bulking sludge from a municipal wastewater treatment plant with anoxic-anaerobic-oxic process was investigated by combination of cultivation and 16S rRNA gene clone library analysis for understanding the causes of bulking.A total of 28 species were obtained from 63 isolates collected from six culture media.The most cultivable species belonged to γ-Proteobacteria including Klebsiella sp.,Pseudomonas sp.,Aeromonas sp.and Acinetobacter sp.Further analysis of these strains by repetitive sequence based on polymerase chain reaction （rep-PCR） technology showed that rep-PCR yielded discriminatory banding patterns within the same genus using REP and BOX primer sets.While the culture-independent assessment revealed that β-Proteobacteria was the dominant group in the bulking sample.Sequence analysis revealed that the highest proportion （14.7%） of operational taxonomic units was 98% similar to Candidatus Accumulibacter phosphatis,which is used to remove phosphorous from wastewater.Our results indicated that combining different approaches can produce complementary information,thus generate a more accurate view of microbial community in bulking sludge.

关键词： bulking sludge cultivable bacteria rep-PCR 16S rRNA gene clone library

Distributed consensus for multiple Euler-Lagrange systems: An event-triggered approach

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Science China(Technological Sciences) 2016年第1期59卷 33-44页

作者： HUANG Na DUAN zhisheng ZHAO yu State Key Laboratory for Turbulence and Complex Systems Department of Mechanics and Engineering Science College of EngineeringPeking University

Distributed consensus problems for multiple Euler-Lagrange systems are addressed on the basis of event-triggered information in this study. Distributed consensus protocols are first designed in terms of two event-triggered scenarios: a decentralized strategy and a distributed strategy. Sufficient conditions that guarantee the event-triggered consensus for multiple Euler-Lagrange systems are then presented, with the associated advantages of reducing controller update times. It is shown that the Zeno behavior of triggering time sequences is excluded for both strategies. Finally, multiple Euler-Lagrange systems that consist of six two-link manipulators are considered to illustrate the effectiveness of the proposed theoretical algorithms.

关键词：拉格朗日系统事件触发分布式欧拉时间相关时间序列理论算法控制器