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柑橘3种病毒类病原多重RT-PCR检测技术的建立及应用

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福建农业学报 2024年第3期39卷 339-344页

作者：袁琳凯马崇欢李丁山陈志炜江宵烽丁新伦张洁吴祖建福建农林大学植物病毒研究所/闽台作物有害生物生态防控国家重点实验室福建福州350002

【目的】建立柑橘黄化脉明病毒(citrus yellow vein clearing virus,CYVCV)、柑橘衰退病毒(citrus tristeza virus,CTV)和啤酒花矮化类病毒(hop stunt viroid,HSVd)的多重RT-PCR检测体系。【方法】设计多重RT-PCR引物,分析其特异性,确定其最佳浓度比、最适退火温度及灵敏度,在此基础上对福建地区的柑橘样品进行检测。【结果】确定了CYVCV-F/R、CTV-F/R和HSVd-F/R等3对引物的最佳浓度比例为1∶1∶2,最适退火温度为52.9℃,灵敏度结果显示该体系可检测模板稀释到10^(-2)的阳性样品。应用该体系对采自福建部分地区的157份柑橘样品进行检测,结果发现,CYVCV、CTV和HSVd的检出率分别为47.1%、56.7%和22.9%。【结论】成功建立了柑橘CYVCV、CTV和HSVd病原的多重RT-PCR检测方法,为该类病害的检测提供准确、快速的检测方法。

关键词：柑橘黄化脉明病毒柑橘衰退病毒啤酒花矮化类病毒多重RT-PCR

Cleavage of the Babuvirus Movement Protein B4 into Functional Peptides Capable of Host Factor Conjugation is Required for Virulence

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Virologica Sinica 2019年第3期34卷 295-305页

作者： Jun Zhuang Wenwu Lin Christopher J.Coates Pengxiang Shang Taiyun Wei zujian wu Lianhui Xie State Key Laboratory of Ecological Pest Control for Fujian and Taiwan Crops Fujian Agriculture and Forestry UniversityFuzhou 350002China Institute of Plant Virology Fujian Agriculture and Forestry UniversityFuzhou 350002China Department of Biosciences College of ScienceSwansea UniversitySwansea SA2 8PPWalesUK

Banana bunchy top virus(BBTV)poses a serious danger to banana crops ***-encoded protein B4 is a determinant of ***,the relevant molecular mechanisms underlying its effects remain *** this study,we found that a functional peptide could be liberated from protein B4,likely via proteolytic ***-directed mutagenesis indicated that the functional processing of protein B4 is required for its pathogenic effects,including dwarfism and sterility,in *** released protein fragment targets host proteins,such as the large subunit of RuBisCO(RbcL)and elongation factor 2(EF2),involved in protein ***,the peptide released from B4(also a precursor)may act as a non-canonical modifier to influence host-pathogen interactions involving BBTV and plants.

关键词： Banana bunchy top virus(BBTV) Movement protein B4 Functional peptide Pathogenicity

Identification of Pns12 as the second silencing suppressor of Rice gall dwarf virus

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Science China(Life Sciences) 2011年第3期54卷 201-208页

作者： wu JianGuo WANG ChunZheng DU ZhengGuo CAI LiJun HU MeiQun wu zujian LI Yi XIE LianHui Institute of Plant Virology Fujian Agriculture and Forestry University Key Laboratory of Plant Virology of Fujian Province Fuzhou 350002 China Peking-Yale Joint Center for Plant Molecular Genetics and Agrobiotechnology National Laboratory of Protein Engineering and Plant Genetic Engineering College of Life Sciences Peking University Beijing 100871 China

RNA silencing is a conserved mechanism found ubiquitously in eukaryotic *** has been used to regulate gene expression and *** addition,RNA silencing serves as an important mechanism in plants' defense against invasive nucleic acids,such as viruses,transposons,and *** a counter-defense,most plants,and some animal viruses,encode RNA silencing suppressors to interfere at one or several points of the silencing *** this study,we showed that Pns12 of RGDV (Rice gall dwarf virus) exhibits silencing suppressor activity on the reporter green fluorescent protein in transgenic Nicotiana benthamiana line ***12 of RGDV suppressed local silencing induced by sense RNA but had no effect on that induced by *** of Pns12 also enhanced Potato virus X pathogenicity in ***,these results suggested that RGDV Pns12 functions as a virus suppressor of RNA silencing,which might target an upstream step of dsRNA formation in the RNA silencing ***,we showed that Pns12 is localized mainly in the nucleus of *** leaf cells.

关键词：水稻瘤矮病毒 RNA沉默双链RNA 鉴定绿色荧光蛋白马铃薯X病毒真核生物基因表达

An optimized strategy of nitrogen-split application based on the leaf positional differences in chlorophyll meter readings

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Journal of Integrative Agriculture 2024年第8期23卷 2605-2617页

作者： Gaozhao wu Xingyu Chen Yuguang Zang Ying Ye Xiaoqing Qian Weiyang Zhang Hao Zhang Lijun Liu zujian Zhang Zhiqin Wang Junfei Gu Jianchang Yang Jiangsu Key Laboratory of Crop Genetics and Physiology/Jiangsu Key Laboratory of Crop Cultivation and Physiology/Agricultural College Yangzhou UniversityYangzhou 225009China Jiangsu Co-Innovation Center for Modern Production Technology of Grain Crops Yangzhou UniversityYangzhou 225009China Key Laboratory of Arable Land Quality Monitoring and Evaluation Ministry of Agriculture and Rural AffairsYangzhou UniversityYangzhou 225127China

Modern rice production faces the dual challenges of increasing grain yields while reducing inputs of chemical ***,the disequilibrium between the nitrogen(N)supplement from the soil and the demand for N of plants is a serious obstacle to achieving these ***-based diagnosis can help farmers make better choices regarding the timing and amount of topdressing N *** objective was to evaluate a non-destructive assessment of rice N demands based on the relative SPAD value(RSPAD)due to leaf positional *** this study,two field experiments were conducted,including a field experiment of different N rates(Exp.I)and an experiment to evaluate the new strategy of nitrogen-split application based on RSPAD(***).The results showed that higher N inputs significantly increased grain yield in modern high yielding super rice,but at the expense of lower nitrogen use efficiency(NUE).The N nutrition index(NNI)can adequately differentiate situations of excessive,optimal,and insufficient N nutrition in rice,and the optimal N rate for modern high yielding rice is higher than conventional *** RSPAD is calculated as the SPAD value of the top fully expanded leaf *** value of the third leaf,which takes into account the non-uniform N distribution within a *** RSPAD can be used as an indicator for higher yield and NUE,and guide better management of N fertilizer ***,we developed a new strategy of nitrogen-split application based on RSPAD,in which the N rate was reduced by 18.7%,yield was increased by 1.7%,and the agronomic N use efficiency was increased by 27.8%,when compared with standard farmers'*** strategy of N fertilization shows great potential for ensuring high yielding and improving NUE at lower N inputs.

关键词： canopy crop management practices nitrogen nitrogen nutrition index rice SPAD

利用酵母双杂交系统筛选本氏烟中与RGSV P5互作的蛋白

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江西农业大学学报 2023年第2期45卷 404-412页

作者：熊桂红胡昱颛吴祖建江西农业大学农学院/江西省薯芋生物学重点实验室江西南昌330045 福建农林大学植物病毒研究所福建福州350002

【目的】水稻草状矮化病是由水稻草状矮化病毒引起的,给粮食产量造成了严重威胁。RGSV P5是该病毒的沉默抑制子,在抵抗寄主RNA沉默中发挥重要作用。筛选、鉴定与水稻草状矮缩病毒(RGSV)沉默抑制子P5互作的寄主蛋白,为揭示水稻草状矮化... 详细信息

【目的】水稻草状矮化病是由水稻草状矮化病毒引起的,给粮食产量造成了严重威胁。RGSV P5是该病毒的沉默抑制子,在抵抗寄主RNA沉默中发挥重要作用。筛选、鉴定与水稻草状矮缩病毒(RGSV)沉默抑制子P5互作的寄主蛋白,为揭示水稻草状矮化病毒致病的分子机制提供理论基础,为该病毒病的防治提供有效策略。【方法】利用酵母双杂交技术筛选与水稻草状矮化病毒P5互作的寄主蛋白。提取感染水稻草状矮化病毒的水稻叶片的总RNA。依据P5基因的CDS序列设计特异性引物。利用RT-PCR技术获得P5基因,将P5基因构建到酵母诱饵表达载体pGBKT7上,利用双酶切鉴定诱饵质粒。将诱饵载体pGBKT7、pGBKT7-P5、重组质粒pGBKT7-P5/pGADT7分别转化到酵母感受态细胞Y2H Gold中,通过观察其在缺陷培养基SD/-Trp、SD/-Leu-Trp/上的生长情况及在SD/-Ade/-His/-Leu/-Trp/X-α-gal上的显色情况检测诱饵质粒的毒性和自激活活性。将烟草酵母cDNA文库质粒转化到含诱饵载体pGBKT7-P5的酵母感受态细胞中,先后用不同缺陷型培养基SD/-Leu/-Trp、SD/-Ade/-His/-Leu/-Trp和SD/-Ade/-His/-Leu/-Trp/X-α-gal筛选后获得阳性克隆。经酵母质粒提取、转化、测序和Blast等分子生物学方法最终获得互作蛋白的序列。【结果】RT-PCR扩增得到P5基因,其大小为576 bp。成功构建诱饵载体pGBKT7-P5。表达的诱饵蛋白P5对酵母菌没有毒性和自激活活性。诱饵蛋白P5从本氏烟酵母cDNA文库中进行大量筛选,获得15个阳性克隆,分析后最终得到7个与P5互作的本氏烟蛋白。经生物信息学分析它们分别为线粒体孔蛋白VDAC、ADP核糖基化因子GTP水解酶激活蛋白ArfGAP、囊泡突触结合蛋白Syt-2、延伸因子eEF1A、脯氨酸合成酶共转录的细菌同源蛋白PROSC、非特征蛋白C9orf78及一种未知蛋白。【结论】本研究成功筛选到7个与水稻草状矮化病毒P5互作的寄主因子。这些互作的寄主因子主要参与转运、生物或非生物胁迫、胁迫应答等生物学过程。深入研究这些互作因子将为解析寄主蛋白参与调控病毒的复制、运动、致病等分子机制提供理论基础,为水稻草状矮化病毒病的防治提供新的思路。

关键词：水稻草状矮化病毒 P5蛋白酵母双杂交蛋白互作

Suppression of Jasmonic Acid-Mediated Defense by Viral-Inducible MicroRNA319 Facilitates Virus Infection in Rice

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Molecular Plant 2016年第9期9卷 1302-1314页

作者： Chao Zhang Zuomei Ding Kangcheng wu Liang Yang Yang Li Zhen Yang Shan Shi Xiaojuan Liu Shanshan Zhao Zhirui Yang Yu Wang Luping Zheng Juan Wei Zhenguo Du Aihong Zhang Hongqin Miao Yi Li zujian wu Jianguo wu Fujian Province Key Laboratory of Plant Virology Institute of Plant Virology Fujian Agriculture and Forestry University Fuzhou Fujian 350002 China The State Key Laboratory of Protein and Plant Gene Research College of Life Sciences Peking University Beijing 100871 China / Plant Protection Institute Hebei Academy of Agriculture and Forestry Sciences Baoding 071000 China These authors contributed equally to this article.

MicroRNAs （miRNAs） are pivotal modulators of plant development and host-virus interactions. However, the roles and action modes of specific miRNAs involved in viral infection and host susceptibility remain largely unclear. In this study, we show that Rice ragged stunt virus （RRSV） infection caused increased accumulation of miR319 but decreased expression of miR319-regulated TCP （TEOSINTE BRANCHED/ CYCLOIDEA/PCF） genes, especially TCP21, in rice plants. Transgenic rice plants overexpressing miP,319 or downregulating TCP21 exhibited disease-like phenotypes and showed significantly higher susceptibility to RRSV in comparison with the wild-type plants. In contrast, only mild disease symptoms were observed in RRSV-infected lines overexpressing TCP21 and especially in the transgenic plants overexpressing miR319- resistant TCP21. Both RRSV infection and overexpression of miR319 caused the decreased endogenous jasmonic acid （JA） levels along with downregulated expression of JA biosynthesis and signaling-related genes in rice. However, treatment of rice plants with methyl jasmonate alleviated disease symptoms caused by RRSV and reduced virus accumulation. Taken together, our results suggest that the induction of miR319 by RRSV infection in rice suppresses JA-mediated defense to facilitate virus infection and symp- tom development.

关键词： miR319 rice ragged stunt virus RRSV TCP21 viral infection jasmonic acid

Ballistic performances of the hourglass lattice sandwich structures under high-velocity fragments

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Defence Technology（防务技术） 2024年第1期31卷 312-325页

作者： He-xiang wu Jia Qu Lin-zhi wu Key Laboratory of Advanced Ship Materials and Mechanics College of Aerospace and Civil EngineeringHarbin Engineering UniversityHarbin150001China School of Civil Engineering Northeast Forestry UniversityHarbin150040China

In this paper,the numerical simulation method is used to study the ballistic performances of hourglass lattice sandwich structures with the same mass under the vertical incidence of *** is paid to elucidating the influences of rod cross-section dimensions,structure height,structure layer,and rod inclination angle on the deformation mode,ballistic performances,and ability to change the ballistic direction of *** results show that the ballistic performances of hourglass lattice sandwich structures are mainly affected by their structural *** this respect,structural parameters optimization of the hourglass lattice sandwich structures enable one to effectively improve their ballistic limit velocity and,consequently,ballistic performances.

关键词： Hourglass lattice sandwich structures Ballistic performances high-velocity Finite element analysis

Resilience-incorporated seismic risk assessment of precast concrete frames with“dry”connections

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Earthquake Engineering and Engineering Vibration 2024年第2期23卷 403-425页

作者： wu Chenhao Tang Yuchuan Cao Xuyang wu Gang Key Laboratory of Concrete and Prestressed Concrete Structures of the Ministry of Education Nanjing 211102China Department of Civil and Environmental Engineering University of CaliforniaLos AngelesCA 90095USA School of Civil Engineering Southeast UniversityNanjing 211102China College of Civil and Transportation Engineering Hohai UniversityNanjing 210024China

A resilience-incorporated risk assessment framework is proposed and demonstrated in this study to manifest the advantageous seismic resilience of precast concrete frame(PCF)structures with“dry”connections in terms of their low damage and rapid *** framework integrates various uncertainties in the seismic hazard,fragility,capacity,demand,loss functions,and post-earthquake *** this study,the PCF structures are distinguished from ordinary reinforced concrete frame(RCF)structures by characterizing multiple limit states for the PCF based on its unique damage ***,probabilistic story-wise pushover analyses are performed to yield story-wise capacities for the predefined limit *** the seismic resilience analysis,a step-wise recovery model is proposed to idealize the functionality recovery process,with separate considerations of the repair and non-repair *** recovery model leverages the economic loss and downtime to delineate the stochastic post-earthquake recovery curves for the resilience loss *** such,contingencies in the probabilistic post-earthquake repairs are incorporated and the empirical judgments on the recovery parameters are largely *** proposed framework is demonstrated through a comparative study between two“dry”connected PCFs and one RCF designed as alternative structural systems for a prototype *** results from the risk quantification indicate that the PCFs show reduced loss hazards and lower expected losses relative to the ***,the PCF equipped with energy dissipation devices at the“dry”connections largely reduces the expected economic loss,downtime,and resilience loss by 29%,56%,and 60%,respectively,compared to the RCF.

关键词： precast concrete frame non-emulative precast system seismic resilience seismic risk functional recovery

New findings on the influence of El Nino events on tropical cyclone formation

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Science China Earth Sciences 2024年第3期67卷 892-893页

作者： Liguang wu Department of Atmospheric and Oceanic Sciences Institute of Atmospheric SciencesFudan UniversityShanghai 200438China

Tropical cyclones typically form over warm tropical and subtropical ocean surfaces, as their formation and intensification rely primarily on the energy provided by the ocean. Pioneering research by Gray (1968) revealed that the formation of tropical cyclones requires the sea surface temperature (SST) to be no less than 26.5℃. Consequently, SST is commonly recognized as a crucial parameter among the environmental factors influencing tropical cyclone formation(Gray, 1979;Emanuel and Nolan, 2004).

关键词： tropical cyclone ocean