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Targeted Mutagenesis of NAC Transcription Factor Gene,OsN AC041,Leading to Salt Sensitivity in Rice

Rice science 2019年第2期26卷 98-108,I0007-I0015页

作者： WANG Bo ZHONG Zhaohui ZHANG Huanhuan WANG Xia LIU Binglin yang lijia HAN Xiangyan YU Deshui ZHENG Xuelian WANG Chunguo SONG Wenqin CHEN Chengbin ZHANG Yong College of Life Sciences Nankai University Department of Biotechnology School of Life Sciences and Technology Center for Informational Biology University of Electronic Science and Technology of China

Salinity is a major abiotic stress factor that seriously affects plant growth. Many genes are involved in the response to salt stress with various metabolism pathways. A number of plant transcription factor family genes have been found to be involved in the salt stress response, and NAM, ATAF and CUC(NAC) transcription factors are thought to act as active regulators during abiotic stress, especially salt stress. In this study, we detected a rice NAC transcription factor coding gene, OsNAC041, and confirmed that it influenced the germination of seeds under salt stress and salt tolerance of plants. OsNAC041 was primarily expressed in the leaves and located in the nucleus. Furthermore, the CRISPR/Cas9 method was used to obtain a targeted osnac041 mutant, of which the plant height was higher than that of the wild-type, showing increased salt sensitivity. Moreover, RNA-seq analysis revealed a number of differentially expressed genes(DEGs) involved in several important signaling pathways in the osnac041 mutant. Subsequently, Kyoto Encyclopedia of Genes and Genomes annotation also revealed differential expression of DEGs associated with mitogen-activated protein kinase signaling, peroxisome, eukaryotictype ABC transporters, photosynthesis and plant hormones, which are involved in stress-related signaling pathways. Overall, our study suggested that OsNAC041 was involved in the salt stress response in rice. These findings not only provide empirical evidence of OsNAC041 function, but also provide new insight into its potential application in rice resistance breeding.

关键词： rice NAC transcription factor CRISPR Cas9 RNA-seq salinity

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Nuclear translocation of OsMFT1 that is impeded by OsFTIP1 promotes drought toleranee in rice

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Molecular Plant 2021年第8期14卷 1297-1311页

作者： Ying Chen Jun Shen Liang Zhangi Haoyue Qi lijia yang Huanyu Wang Jiaxuan Wang Yuexing Wang Hao Du Zeng Tao Ting Zhao Pingchuan Deng Qingyao Shu Qian Qian Hao Yu Shiyong Song State Key Laboratory of Rice Biology Zhejiang Provincial Key Laboratory of Crop Genetic ResourcesInstitute of Crop ScienceCollege of Agriculture and BiotechnologyZhejiang UniversityHangzhou 310058China State Key Laboratory of Rice Biology China National Rice Research InstituteHangzhou 311400China lnstitute of Biotechnology College of Agriculture and BiotechnologyZhejiang UniversityHangzhou 310058China State Key Laboratory of Crop Stress Biology in Arid Areas College of AgronomyNorthwest A&F UniversityYanglingShaanxi 712100China Department of Biological Sciences and Temasek Life Sciences Laboratory National University of SingaporeSingapore 117543Singapore

Drought is the leading environmental threat affecting crop productivity,and plants have evolved a series of mechanisms to adapt to drought *** FT-interacting proteins(FTIPs)and phosphatidylethanola *** proteins(PEBPs)play key roles in developmental processes,whereas their roles in the regulation of stress response are still largely ***,we report that OsFTIP1 negatively regulates drought response in *** showed that OsFTIP1 interacts with rice MOTHER OF FT AND TFL1(OsMFT1),a PEBP that promotes rice tolerance to drought *** studies discovered that OsMFT1 interacts with two key drought-related transcription factors,OsbZIP66 and OsMYB26,regulating their binding capacity on drought-related genes and thereby enhancing drought toleranee in ***,we found that OsFTIP1 impedes the nucleocytoplasmic translocation of OsMFT1,implying that dynamic modulation of drought-responsive genes by the OsMFT1-OsMYB26 and OsMFT1-OsbZIP66 complexes is integral to OsFTIP1-modulated nuclear accumulation of *** findings also suggest that OsMFT1 might act as a hitherto unknown nucleocytoplasmic trafficking signal that regulates drought tolerance in rice in response to environmental signals.

关键词： OsFTIPI OsMFTI drought response nucleocytoplasmic trafficking rice

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The OsFTIP6-OsHB22-OsMYBR57 module regulates drought response in rice

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Molecular Plant 2022年第7期15卷 1227-1242页

作者： lijia yang Ying Chen Liang Xu Jiaxuan Wang Haoyue Qi Jiazhuo Guo Liang Zhang Jun Shen Huanyu Wang Fan Zhang Lijun Xie Wenjun Zhu Peitao Lü Qian Qian Hao Yu Shiyong Song State Key Laboratory of Rice Biology Zhejiang Provincial Key Laboratory of Crop Genetic ResourcesInstitute of Crop ScienceCollege of Agriculture and BiotechnologyZhejiang UniversityHangzhou 310058China State Key Laboratory of Rice Biology China National Rice Research InstituteHangzhou 311400China College of Horticulture FAFU-UCR Joint Center for Horticultural Biology and MetabolomicsHaixia Institute of Science and TechnologyFujian Agriculture and Forestry UniversityFuzhou 350002China Department of Biological Sciences and Temasek Life Sciences Laboratory National University of SingaporeSingapore 117543Singapore

Plants have evolved a sophisticated set of mechanisms to adapt to drought *** factors play crucial roles in plant responses to various environmental stimuli by modulating the expression of numerous stress-responsive ***,how the crosstalk between different transcription factor families orchestrates initiation of the key transcriptional network and the role of posttranscriptional modification of transcription factors,especially in cellular localization/trafficking in response to stress in rice,remain still largely *** this study,we isolated an Osmybr57 mutant that displays a drought-sensitive phenotype through a genetic screen for drought stress *** found that OsMYBR57,an MYB-related protein,directly regulates the expression of several key drought-related OsbZ/Ps in response to drought *** studies revealed that OsMYBR57 interacts with a homeodomain transcription factor,OsHB22,which also plays a positive role in drought *** further demonstrate that OsFTIP6 interacts with OsHB22 and promotes the nucleocytoplasmic translocation of OsHB22 into the nucleus,where OsHB22 cooperates with OsMYBR57 to regulate the expression of drought-responsive *** findings have revealed a mechanistic framework underlying the OsFTIP6-0sHB22-0sMYBR57 module-mediated regulation of drought response in *** OsFTIP6-mediated OsHB22 nucleocytoplasmic shuttling and OsMYBR57-0sHB22 regulation of OsbZIP transcription ensure precise control of expression of OsLEA3 and Rab21,and thereby regulate the response to water deficiency in rice.

关键词： OsMYBR57 OsHB22 OsFTIP6 drought response nucleocytoplasmic trafficking rice

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Plant Genome Editing Using FnCpfl and LbCpf Nucleases at Redefined and Altered PAM-Sites

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Molecular Plant 2018年第7期11卷 999-1002页

作者： ZhaohuiZhong Yingxiao Zhang Qi You XU Tang Qiurong Ren Shishi Liu lijia yang Yan Wang Xiaopei Liu Binglin Liu Tao Zhang Xuelian Zheng Ysa Le Yong Zhang Yiping Qi Department of Biotechnology School of Life Sciences and Technology Center for Informational Biology University of Electronic Science andTechnology of China Room 216 Main Building No. 4 Section 2 North J~anshe Road Chengdu 610054 P.R. China Department of Plant Science and Landscape Architecture University of Maryland College Park MD 20742 USA Jiangsu Key Laboratory of Crop Genetics and Physiology Co-InnovationCenter for Modern Production Technology of Grain Crops Key Laboratory ofPlant Functional Genomics of the Ministry of Education Yangzhou University Yangzhou 225009 China Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China Yangzhou University Yangzhou 225009 China Institute for Bioscience and Biotechnology Research University of Maryland Rockville MD 20850 USA These authors contributed equally to this article.

Dear Editor CRISPR from Prevotella and Francisella 1 （Cpfl） is an emerging RNA-guided endonuclease system that relies on thymidine-rich protospacer adjacent motif （PAM） for DNA targeting （Zetsche et al., 2015）. CRISPR-Cpfl has unique features that could be advantageous over the CRISPR-Cas9 system. For example, Cpfl requires only a 42 nt crRNA, while Cas9 uses 100 nt gRNA. While Cas9 generates blunt ends of DNA breaks, the Cpfl cleavage results in 5＇ overhangs distal from the protospacer, which may improve efficiency for NHEJ-based gene insertion. Interestingly, Cpfl proteins also have RNase activity （Fonfara et al., 2016）, which was utilized to process crRNA arrays for multiplexed genome editing in both mammalian systems and plants （Wang et al., 2017; Zetsche et al., 2017）.

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Improving Plant Genome Editing with High-Fidelity xCas9 and Non-canonical PAM-Targeting Cas9-NG

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Molecular Plant 2019年第7期12卷 1027-1036页

作者： Zhaohui Zhong Simon Sretenovic Qiurong Ren lijia yang Yu Bao Caiyan Qi Mingzhu Yuan Yao He Shishi Liu Xiaopei Liu Jiaheng Wang Lan Huang Yan Wang Dibin Baby David Wang Tao Zhang Yiping Qi Yong Zhang Department of Biotechnology School of Life Sciences and Technology Center for Informational Biology University of 曰ectronic Science and Technology of China Room 216 Main Building No. 4 Section 2 North Jianshe Road Chengdu 610054 China Department of Plant Science and Landscape Architecture University of Maryland College Park MD 20742 USA Jiangsu Key Laboratory of Crop Genetics and Physiology Co-Innovation Center for Modern Production Technology of Grain Crops Key Laboratory of Plant Functional Genomics of the Ministry of Education Yangzhou University Yangzhou 225009 China Joint International Research Laboratory of Agriculture and Agri-Product Safety of Ministry of Education of China Yangzhou University Yangzhou 225009 China Indian Institute of Science Education and Research Timpati Andhra Pradesh 517507 India Seven Lakes High School Katy TX 77494 USA Institute for Bioscience and Biotechnology Research University of Maryland Rockville MD 20850 USA

Two recently engineered SpCas9 variants, namely xCas9 and Cas9-NG, show promising potential in improving targeting specificity and broadening the targeting range. In this study, we evaluated these Cas9 variants in the model and crop plant, rice. We first tested xCas9-3.7, the most effective xCas9 variant in mammalian cells, for targeted mutagenesis at 16 possible NGN PAM (protospacer adjacent motif) combinations in duplicates. xCas9 exhibited nearly equivalent editing efficiency to wild-type Cas9 (Cas9-WT) at most canonical NGG PAM sites tested, whereas it showed limited activity at non-canonical NGH (H = A, C, T) PAM sites. High editing efficiency of xCas9 at NGG PAMs was further demonstrated with C to T base editing by both rAPOBECI and PmCDAI cytidine deaminases. With mismatched sgRNAs, we found that xCas9 had improved targeting specificity over the Cas9-WT. Furthermore, we tested two Cas9-NG variants, Cas9-NGv1 and Cas9-NG, for targeting NGN PAMs. Both Cas9-NG variants showed higher editing efficiency at most non-canonical NG PAM sites tested, and enabled much more efficient editing than xCas9 at AT-rich PAM sites such as GAT, GAA, and CAA. Nevertheless, we found that Cas9-NG variants showed significant reduced activity at the canonical NGG PAM sites. In stable transgenic rice lines, we demonstrated that Cas9-NG had much higher editing efficiency than Cas9-NGv1 and xCas9 at NG PAM sites. To expand the base-editing scope, we developed an efficient C to T base-editing system by making fusion of Cas9-NG nickase (D10A version), PmCDAI, and UGI. Taken together, our work benchmarked xCas9 as a high-fidelity nuclease for targeting canonical NGG PAMs and Cas9-NG as a preferred variant for targeting relaxed PAMs for plant genome editing.

关键词： xCas9 Cas9-NG high fidelity non-canonical PAM base editing

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2013-2018年中国结核病空间聚集性分析

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中华流行病学杂志 2020年第11期41卷 1843-1847页

作者：杨丽佳李涛陈伟中国疾病预防控制中心结核病预防控制中心北京102206

目的分析2013-2018年中国结核病以县(区)为单位的空间聚集性特征及其变化趋势。方法数据来源于中国疾病预防控制信息系统结核病信息管理系统,以2013-2018年全国各县(区)结核病登记数据和同期人口数据为基础,描述2013-2018年结核病疫情... 详细信息

目的分析2013-2018年中国结核病以县(区)为单位的空间聚集性特征及其变化趋势。方法数据来源于中国疾病预防控制信息系统结核病信息管理系统,以2013-2018年全国各县(区)结核病登记数据和同期人口数据为基础,描述2013-2018年结核病疫情的空间分布特征,采用SaTScan 9.6软件对2013-2018年以县(区)为单位的结核病登记率进行空间扫描,通过扫描窗口下的对数似然比(LLR)和P值判断结核病聚集区的位置和范围,在此基础上分析聚集区结核病发病的RR值。结果2013-2018年,全国结核病登记率基本呈缓慢下降趋势。全国范围内存在多个空间高值聚集区,主要位于西部的新疆维吾尔自治区(新疆)、西藏自治区(西藏)、陕西省、四川省、重庆市、云南省、贵州省、广西壮族自治区(广西),中东部的内蒙古自治区、湖北省、湖南省、江西省、广东省、海南省。结核病空间聚集区覆盖的县(区)数量呈缓慢下降趋势,从2013年的1112个下降到2018年的806个。新疆南疆地区、西藏西部地区的结核病聚集区相比于其他地区,结核病发病的RR值始终维持在较高水平(RR>3.0)。结论2013-2018年我国结核病聚集区域覆盖的县(区)数量逐渐减少,聚集区主要分布于西部、中部的多个省份,应加强结核病聚集区的结核病患者健康教育和早期发现等防控工作,防止结核病疫情的扩散。西部地区不仅存在结核病聚集性,且结核病发病RR值较高,应加强结核病患者的治疗管理等工作。

关键词：结核病空间聚集性分析

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“岗课证融通育训赛一体”实践教学体系研究——以现代物流管理专业为例

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天津职业院校联合学报 2023年第3期25卷 36-41,49页

作者：杨立佳天津交通职业学院教务部(项目办) 天津300110

立足于新时代物流人才培养需求,着眼于职业教育“岗课赛证”融合育人内涵提升需要,以天津交通职业学院现代物流管理专业实践教学体系建设经验为蓝本,开展“岗课证融通育训赛一体”标准化实践教学体系建设研究。聚焦于现代物流管理专业... 详细信息

立足于新时代物流人才培养需求,着眼于职业教育“岗课赛证”融合育人内涵提升需要,以天津交通职业学院现代物流管理专业实践教学体系建设经验为蓝本,开展“岗课证融通育训赛一体”标准化实践教学体系建设研究。聚焦于现代物流管理专业职业教育标准化研究,落实立德树人根本任务,创建了“四级育训融合”现代物流实践教学标准;以实践育人为目标,打造了“行企校定岗岗课证融通”的现代物流实践育人模式,培育新时代物流工匠;以实践教学管理机制、实践教学条件、实践教学资源、实践教学评价体系建设为抓手,形成了“产学创融合育训赛一体”的现代物流实践教学实施方案,培养复合型技术技能人才,服务物流产业链发展。

关键词：现代物流管理专业标准化实践教学体系岗课赛证

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深化“最多跑一次”改革的实践和探索——以杭州公安交管部门实施“最多跑一次”改革为例

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浙江警察学院学报 2019年第4期31卷 112-120页

作者：杨力加杭州市公安局

交通警察是公安机关接触群众最多的警种之一,也是"最多跑一次"改革的重点警种。自"最多跑一次"改革开展以来,杭州公安交管部门为切实推进"简政放权、放管结合、优化服务"的改革部署,以"规范办、简化办、网上办、就近办"为方向,从"简政... 详细信息

交通警察是公安机关接触群众最多的警种之一,也是"最多跑一次"改革的重点警种。自"最多跑一次"改革开展以来,杭州公安交管部门为切实推进"简政放权、放管结合、优化服务"的改革部署,以"规范办、简化办、网上办、就近办"为方向,从"简政放权、数据跑腿、举措创新、机制保障"四个方面着力,形成了一套完整的改革链条,摸索出了杭州交警改革的路子,较好地实现了让群众"少跑""跑好"。进一步深化"最多跑一次"改革,应当以更加鲜明的问题导向促进人民群众诉求的全面满足,以更加深入的流程再造推动改革举措落地抓实,以更大程度的数据共享推动构建部门在线协作机制,以更大力度的保障机制拓宽改革成效的巩固升华。

关键词： “最多跑一次” 行政审批制度 “放管服” 公安交通管理杭州市公安局

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基于消费者购买历史的平台企业价格决策

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系统管理学报 2023年第5期32卷 939-959页

作者：董维刚杨立佳闫文璐大连理工大学经济管理学院辽宁大连116024

大数据时代,平台企业通过对消费者画像实现针对不同类型消费者的个性化产品推荐和差别定价的现象屡屡发生。将企业的这种精准推荐产品和服务行为以消费者效用的形式纳入豪特林模型中,分析双寡头企业进行差别定价的动机并比较分析不同定... 详细信息

大数据时代,平台企业通过对消费者画像实现针对不同类型消费者的个性化产品推荐和差别定价的现象屡屡发生。将企业的这种精准推荐产品和服务行为以消费者效用的形式纳入豪特林模型中,分析双寡头企业进行差别定价的动机并比较分析不同定价方式下的福利。研究结果表明:在双寡头市场竞争中,与统一定价相比,企业的差别定价行为会降低企业利润,但能提高企业的市场份额,当双寡头企业采取不对称定价策略时,进行差别定价的一方在市场份额竞争中取得优势;当双寡头企业采取有技术差距的差别定价时,技术领先的一方在市场份额竞争中取得优势。与直觉相反,研究表明,差别定价并不总是损害消费者福利。由此,有关部门不应盲目干预差别定价行为,但要关注企业通过差别定价进行无序扩张行为,从而避免企业形成市场支配地位后进行不正当竞争、谋取利益等乱象的发生。

关键词：平台企业差别定价精准服务豪特林模型

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电磁干扰及抑制分析

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电子测试 2019年第4期30卷 35-37,54页

作者：杨礼迦广西玉林市机电工程学校广西玉林537000

本文分析了电磁干扰的途径和形式。

关键词：电磁干扰抑制滤波屏蔽接地布线

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