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Robust Transcriptional Activation in Plants Using Multiplexed CRISPR-Act2.0 and mTALE-Act systems

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Molecular Plant 2018年第2期11卷 245-256页

作者： Levi G. Lowder Jianping Zhou Yingxiao Zhang Aimee Malzahn Zhaohui Zhong Tzung-Fu Hsieh Daniel F. Voytas Yong Zhang Yiping Qi department of Biology East Carolina University Greenville NC 27858 USA department of Biotechnology School of Life Science and Technology Center for informational Biology University of Electronic Science and Technology of ChinaChengdu 610054 China department of Plant Science and landscape architecture University of Maryland College Park MD 20742 USA department of Plant and Microbial Biology and Plants for Human Health Institute North Carolina State University North Carolina Research Campus KannapolisNC 28O81 USA department of Genetics Ceil Biology and Development Center for Genome Engineering University of Minnesota Minneapolis MN 55455 USA Institute for Bioscience and Biotechnology Research University of Maryland Rockville MD 20850 USA

User-friendly tools for robust transcriptional activation of endogenous genes are highly demanded in plants. We previously showed that a dCas9-VP64 system consisting of the deactivated CRISPR- associated protein 9 （dCasg） fused with four tandem repeats of the transcriptional activator VP16 0/1=64） could be used for transcriptional activation of endogenous genes in plants. In this study, we developed a second generation of vector systems for enhanced transcriptional activation in plants. We tested multiple strategies for dCasg-based transcriptional activation, and found that simultaneous recruitment of VP64 by dCas9 and a modified guide RNA scaffold gRNA2.0 （designated CRISPR-Act2.0） yielded stronger transcrip- tional activation than the dCas9-VP64 system. Moreover, we developed a multiplex transcription activator- likeeffector activation （mTALE-Act） system for simultaneous activation of up to four genes in plants. Our results suggest that mTALE-Act is even more effective than CRISPR-Act2.0 in most cases tested. In addition, we explored tissue-specific gene activation using positive feedback loops. Interestingly, our study revealed that certain endogenous genes are more amenable than others to transcriptional activation, and tightly regulated genes may cause target gene silencing when perturbed by activation probes. Hence, these new tools could be used to investigate gene regulatory networks and their control mechanisms. Assembly of multiplex CRISPR-Act2.0 and mTALE-Act systems are both based on streamlined and PCR-independent Golden Gate and Gateway cloning strategies, which will facilitate transcriptional activation applications in both dicots and monocots.

关键词： CRISPR gRNA2.0 MS2-VP64 TALE-VP64 multiplex transcriptional activation

Internet of Things Based Solutions for Transport Network Vulnerability Assessment in Intelligent Transportation systems

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Computers, Materials & Continua 2020年第12期65卷 2511-2527页

作者： Weiwei Liu Yang Tang Fei Yang Chennan Zhang Dun Cao Gwang-jun Kim Business School University of Shanghai for Science and TechnologyShanghai200093China Zhejiang University Urban and rural Planning&Design Institute Hangzhou310023China School of Computer&Communication engineering Changsha University of Science&TechnologyChangsha410004China department of Computer engineering Chonnam National UniversityGwangju61186Korea

Intelligent Transportation system(ITS)is essential for effective identification of vulnerable units in the transport network and its stable ***,it is necessary to establish an urban transport network vulnerability assessment model with solutions based on Internet of Things(IoT).Previous research on vulnerability has no congestion effect on the peak time of urban road *** cascading failure of links or nodes is presented by IoT monitoring system,which can collect data from a wireless sensor network in the transport *** IoT monitoring system collects wireless data via Vehicle-to-Infrastructure(V2I)channels to simulate key segments and their failure ***,the topological structure vulnerability index and the traffic function vulnerability index of road network are extracted from the vulnerability *** two indices are standardized by calculating the relative change rate,and the comprehensive index of the consequence after road network unit is in a failure ***,by calculating the failure probability of road network unit and comprehensive index of road network unit in failure state,the comprehensive vulnerability of road network can be evaluated by a risk calculation *** short,the IoT-based solutions to the new vulnerability assessment can help road network planning and traffic management departments to achieve the ITS goals.

关键词： Internet of Things Intelligent Transport systems vulnerability assessment transport network

Carbon stock estimation by dual-polarized synthetic aperture radar (SAR) and forest inventory data in a Mediterranean forest landscape

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Journal of Forestry Research 2022年第3期33卷 827-838页

作者： Can Vatandaşlar Saygin Abdikan department of Forest engineering Artvin Coruh University08100ArtvinTurkey department of Geomatics engineering Hacettepe University06800AnkaraTurkey

Forest ecosystems play a crucial role in mitigating global climate change by forming massive carbon sinks. Their carbon stocks and stock changes need to be quantified for carbon budget balancing and international reporting schemes. However, direct sampling and biomass weighing may not always be possible for quantification studies conducted in large forests. In these cases, indirect methods that use forest inventory information combined with remote sensing data can be beneficial. Synthetic aperture radar (SAR) images offer numerous opportunities to researchers as freely distributed remote sensing data. This study aims to estimate the amount of total carbon stock (TCS) in forested lands of the Kizildag Forest Enterprise. To this end, the actual storage capacities of five carbon pools, i.e. above- and below-ground, deadwood, litter, and soil, were calculated using the indirect method based on ground measurements of 264 forest inventory plots. They were then associated with the backscattered values from Sentinel-1 and ALOS-2 PALSAR-2 data in a Geographical Information system (GIS). Finally, TCS was separately modelled and mapped. The best regression model was developed using the HH polarization of ALOS-2 PALSAR-2 with an adjusted R^(2) of 0.78 (p systems could be estimated using appropriate SAR images at acceptable accuracy levels for forestry purposes. The use of additional ancillary data may provide more delicate and reliable estimations in the future. Given the implications of this study, the spatiotemporal dynamics of carbon can be effectively controlled

关键词： Carbon storage Aboveground carbon Soil-bound carbon Forest biomass Synthetic aperture radar(SAR)

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DNA纳米技术应用于水环境污染示踪

科学通报 2024年第1期69卷 112-126页

作者：王常茜仰大勇罗丹廖人宽中国农业大学土地科学与技术学院北京100193 农业农村部华北耕地保育重点实验室北京100193 复旦大学化学与材料学院上海200438 department of Biological and Environmental engineering Cornell UniversityIthaca 14853USA

水环境污染是当前人类社会所面临的严峻挑战之一,阻碍了全球生态环境和社会经济的可持续发展.潜在污染源众多和水文地质条件的复杂性导致对污染源追溯、污染范围界定和污染程度量化等问题的研判十分棘手.针对这些难题,已发展了多种水环... 详细信息

水环境污染是当前人类社会所面临的严峻挑战之一,阻碍了全球生态环境和社会经济的可持续发展.潜在污染源众多和水文地质条件的复杂性导致对污染源追溯、污染范围界定和污染程度量化等问题的研判十分棘手.针对这些难题,已发展了多种水环境污染物迁移示踪系统,例如离子化合物、有色染料和同位素等,这些示踪系统的应用对水环境污染治理起到了重要的推动作用.近年来,快速发展的基于脱氧核糖核酸(deoxyribo nucleic acid,DNA)纳米技术的示踪方法(DNA示踪技术)逐渐崭露头角,其是应用DNA片段作为特异性标记物进行水环境污染示踪.与已有的示踪技术相比,DNA示踪技术具有示踪剂数量巨大、特异性强、检测灵敏、运移及降解特性可控和环境友好等诸多优势,是极具潜力的新一代示踪技术.DNA示踪技术的研发与应用集成了生物化学、材料科学、环境工程和水文地质等多学科知识,是典型的交叉研究领域.为促进DNA示踪技术体系的长足发展以及水环境污染防治能力的不断增强,本文重点阐述了DNA片段作为水环境污染示踪剂的原理与方法,全面梳理了DNA示踪技术体系的发展脉络,总结探讨了DNA示踪技术在水环境污染示踪研究中的典型应用,最后展望了未来重要的研究方向.

关键词：生物纳米技术基因序列示踪剂污染物运移水环境保护

Urban Utopia and Building Experimentations： The rural Villages of the Sicilian Latifundium

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Journal of Civil engineering and architecture 2014年第9期8卷 1138-1144页

作者： Vincenzo Sapienza department of Civil engineering and architecture Catania University Catania 95125 Italy

The colonization of Sicilian latifundium was an attempt of regeneration of the rural areas. It was carried out by Benito Mussolini, called II Duce, who governed Italy by a regime with an absolute and conservative approach for 20 years since 1922. According to this plan, there were some rural villages （with a school, a church, a police station and a doctor＇s surgery） and a number of rural homes, spread in the landscape, that formed one of the most important achievements of the agrarian reform in Italy. The author has been conducting scientific research on this topic for several years. He started with the archive ＂Ente per la Colonizzazione del Latifondo Siciliano＂（now called Ente Sviluppo Agricolo） to review the original designs and contracts with companies, ledgers and other administrative documents. Lately, he has been reviewing the personal archives of designers involved in the building program, where he has found original drawings, letters, sketches, etc.. He has also conducted some architectonic and technical surveys of the sites. We have to consider that nowadays the mentioned rural villages are a strategic resource for the development of the neighboring areas, as it may become a network for the sustainable tourism and the promotion of the local agro food specialties. On this purpose, the local authority is going to promote some plans for the requalification of these sites. The particular achievement of this article is to show a number of possibilities for the reuse of these places.

关键词： rural village autarchy construction history rehabilitation.

Application of 3D bioprinting in the prevention and the therapy for human diseases

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Signal Transduction and Targeted Therapy 2021年第6期6卷 1681-1697页

作者： Hee-Gyeong Yi Hyeonji Kim Junyoung Kwon Yeong-Jin Choi Jinah Jang Dong-Woo Cho department of rural and Biosystems engineering College of Agriculture and Life SciencesChonnam National University77 Yongbong-RoGwangju 61186Korea department of Mechanical engineering Pohang University of Science and Technology(POSTECH)77 Cheongam-RoPohangKyungbuk 37673Korea department of Advanced Biomaterials Research Korea Institute of Materials Science(KIMS)797 ChangwondaeroChangwonKyungnam 51508Korea department of Convergence IT engineering POSTECH77 Cheongam-RoPohangKyungbuk 37673Korea Institute of Convergence Science Yonsei University50 Yonsei-RoSeoul 03722Korea

Rapid development of vaccines and therapeutics is necessary to tackle the emergence of new pathogens and infectious *** speed up the drug discovery process,the conventional development pipeline can be retooled by introducing advanced in vitro models as alternatives to conventional infectious disease models and by employing advanced technology for the production of medicine and cell/drug delivery *** this regard,layer-by-layer construction with a 3D bioprinting system or other technologies provides a beneficial method for developing highly biomimetic and reliable in vitro models for infectious disease *** addition,the high flexibility and versatility of 3D bioprinting offer advantages in the effective production of vaccines,therapeutics,and relevant delivery ***,we discuss the potential of 3D bioprinting technologies for the control of infectious *** also suggest that 3D bioprinting in infectious disease research and drug development could be a significant platform technology for the rapid and automated production of tissue/organ models and medicines in the near future.

关键词： printing prevention introducing

Linear diophantine uncertain linguistic-based prospect theory approach for performance evaluation of islanded microgrid-system scenarios

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Clean Energy 2023年第2期7卷 263-282页

作者： Sweta Singh Neeraj Kanwar Divya Zindani department of Electrical engineering Manipal University JaipurIndia department of Mechanical engineering Sri Sivasubramaniya Nadar College of EngineeringKalavakkamTamil NaduIndia

Renewable-energy-based hybrid microgrids can aid in achieving one of the United Nations Sustainable Development Goals,i.e.‘Affordable and clean energy’.However,experts may be faced with the challenge of selecting the best one for the electrification of an *** avoid the challenge and realize the ultimate goal of the United Nations,the present study,therefore,proposes a novel pros-pect theory-based decision-making approach to help experts in opting for the best microgrid *** proposed decision-making framework considers the risk appetite of the decision-maker,a quintessential aspect of the *** diophantine uncertain lin-guistic sets are used to model the linguistic evaluations from the *** information from different experts is aggregated using a linear diophantine uncertain linguistic power Einstein-weighted geometric ***,the prospect-theory-based TOmada de Decisao Interativa Multicriterio approach is employed to evaluate the performance of the available microgrid scenarios and hence opt for the best microgrid *** proposed framework has been used to evaluate the performance of seven possible microgrid scenarios and hence select the best one that can be implemented for rural electrification of a remote village in Assam,*** microgrid scenario consisting of a photovoltaic-wind turbine-fuel cell-battery converter(MG_(3))has been revealed to be the best scen-ario among the seven considered microgrid *** validity of the obtained ranking results has been adjudged through a com-prehensive evaluation regarding the attenuation factor and the weights of the ***,previous case studies have also been solved using the proposed methodology and the results reveal a good correlation between the obtained ranking results.

关键词： distributed energy and smart grid energy and environment islanded microgrid

An optimized solar-air degree-day method to evaluate energy demand for poultry buildings in different climate zones

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Frontiers of Agricultural Science and engineering 2020年第4期7卷 478-489页

作者： Yang WANG Baoming LI department of Agricultural Structure and Bioenvironmental engineering College of Water Resources and Civil EngineeringChina Agricultural UniversityBeijing 100083China Laboratory of Agricultural engineering in Structure and Environment Ministry of Agriculture and Rural AffairsBeijing 100083China Beijing engineering Research Center for Animal Healthy Environment Beijing 100083China

The degree-day method is widely used to determine energy consumption but cannot be directly applied to poultry buildings without improvements in its *** study was designed to optimize the degreeday calculation and proposes a solar-air degree-day method,which can be used to calculate the cooling and heating degree-days and the annual cooling and heating loads under different climate conditions for poultry *** this paper,the solar-air degree-day method was proposed,which considers the effects of solar radiation with different wall orientations and surface *** Chinese cities,Harbin,Beijing,Chongqing,Kunming and Guangzhou,were selected to represent different climate zones to determine the solar-air *** heating and cooling energy requirements for different climates were compared by DeST(Designer’s Simulation Toolkit)simulation and the solar-air degreeday *** to decrease energy consumption were *** results showed that the maximum relative error was less than 10%,and the new method was not significantly different from the DeST simulation(P>0.05).The accuracy of calculating energy requirements was improved by the solar-air degree-day method in the different climate *** and surface color effects on energy consumption need to be considered,and external walls of different orientations should have different surface colors.

关键词： base temperature energy consumption solar radiation orientation surface color

Exogenous SA Initiated Defense Response and Multi-signaling Pathway in Tetraploid Potato SD20

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Horticultural Plant Journal 2020年第2期6卷 99-110页

作者： Jiayi Zheng Yu Yang Xiao Guo Liping Jin Xingyao Xiong Xiaohui Yang Guangcun Li Key Laboratory of Biology and Genetic Improvement of Tuber and Root Crop Institute of Vegetables and FlowersChinese Academy of Agricultural SciencesMinistry of Agriculture and Rural AffairsBeijing 100081China National Vegetable Improvement Center Shandong Sub-Center/Huang-Huai-Hai Region Scientific Observation and Experimental Station of Vegetables Ministry of Agriculture and Rural Affairs/Molecular Biology Key Laboratory of Shandong Facility VegetableInstitute of Vegetables and FlowersShandong Academy of Agricultural SciencesJinan 250100China Hu’nan Provincial engineering Research Center for Potatoes College of Horticulture and LandscapeHunan Agricultural UniversityChangsha 410128China

Salicylic acid(SA) is an important signaling substance that plays an important role in plant growth, development and disease *** order to further understand the role of the SA pathway in potato disease resistance and identify SA signaling key genes, gene expression profiling of the late blight resistance genotype SD20 was performed under exogenous SA application. A total of 28 572 unigenes were assembled,of which 4 564 were differentially expressed. Analysis of differentially expressed genes(DEGs) showed that multiple signaling pathways such as SA, jasmonic acid, ethylene, abscisic acid, auxin, and brassinolide were involved in response to exogenous SA. Many plant defense signalrelated genes involved in protein serine/threonine kinase activity and plant-pathogen interaction, were significantly enriched. These were consistent with the interaction results of SD20 and Phytophthora infestans in our previous study, indicating that exogenous SA stimulated the resistance response and initiated a similar defense pathway compared to pathogen infection in SD20, which confirmed crosstalk of the SA signaling pathway with a pathogen-induced disease resistance signal pathway in plant. Moreover, transcriptome analysis revealed that ROS1 was positively regulated by SA in potato for the first time. SA-induced gene expression profiling provides insight into SA signaling and its mechanisms in disease defense systems.

关键词： potato salicylic acid transcriptome pathway defense response