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Development of a stable attenuated double-mutant of tobacco mosaic virus for cross-protection

Journal of Integrative Agriculture 2024年第7期23卷 2318-2331页

作者： Xiaojie Xu Shaoyan Jiang Chunju Liu Xujie Sun Qing Zhu Xiuzhai Chen Pengchao Jiang Fenglong Wang Yanping Tian Xiangdong Li The Engineering Research Institute of Agriculture and Forestry Ludong UniversityYantai 264025China Laboratory of Plant Virology Department of Plant PathologyCollege of Plant ProtectionShandong Agricultural UniversityTai’an 271018China Weifang tobacco Corporation Weifang 261031China Linyi tobacco Corporation Linyi 276000China tobacco Research Institute Chinese Academy of Agricultural SciencesQingdao 266101China

tobacco(Nicotiana tabacum)and tomato(Solanum lycopersicum)are two major economic crops in China.tobacco mosaic virus(TMV;genus Tobamovirus)is the most prevalent virus infecting both crops.Currently,some widely cultivated tobacco and tomato cultivars are susceptible to TMV and there is no effective strategy to control this virus.Cross-protection can be a safe and environmentally friendly strategy to prevent viral diseases.However,stable attenuated TMV mutants are scarce.In this study,we found that the substitutions in the replicase p126,arginine at position 196(R196)with aspartic acid(D),glutamic acid at position 614(E614)with glycine(G),serine at position 643(S643)with phenylalanine(F),or D at position 730(D730)with S,significantly reduced the virulence and replication of TMV.However,only the mutation of S643 to F reduced the RNA silencing suppression activity of TMV p126.A double-mutant TMV-E614G-S643F induced no visible symptom and was genetically stable through six successive passages in tobacco plants.Furthermore,our results showed that TMV-E614G-S643F double-mutant could provide effective protection against the wild-type TMV infection in tobacco and tomato plants.This study reports a promising mild mutant for cross-protection to control TMV in tobacco and tomato plants.

关键词： cross-protection double-mutant p126 tobacco mosaic virus virulence

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A Viral Protein Suppresses siRNA-directed Interference in tobacco mosaic virus Infection

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Acta Biochimica et Biophysica Sinica 2005年第4期37卷 248-253页

作者： Ming-Min ZHAO De-Rong AN Guang-Hua HUANG~1 Zu-Hua HE~2 Jiang-Ye CHEN~1 College of Plant Protection,North West Science and Technology University of Agriculture and Forestry,Shaanxi 712100,China ~1State Key Laboratory of Molecular Biology,Institute of Biochemistry and Cell Biology,Shanghai Institutes for Biological Sciences. Chinese Academy of Sciences,Shanghai 200031,China ~2State Key Laboratory of Plant Molecular Genetics.Institute of Plant Physiology and Ecology,Shanghai Institutes for Biological Sciences.Chinese Academy of Sciences,Shanghai 200032.China College of Plant Protection North West Science and Technology University of Agriculture and ForestryShaanxi 712100 China State Key Laboratory of Molecular Biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of SciencesShanghai 200031 China State Key Laboratory of Plant Molecular Genetics Institute of Plant Physiology and Ecology Shanghai Institutes for Biological Sciences Chinese Academy of SciencesShanghai 200032 China

Plant viruses encode suppressors of post-transcriptional gene silencing (PTGS), an adaptivedefense response that limits virus replication and its spread in plants.The helper component proteinase (HC-Pro) of the potato virus A (PVA,genus Potyvirus) suppresses PTGS of silenced transgenes.Here,the effectof HC-Pro on siRNA-directed interference in the tobacco mosaic virus (TMV) was examined by using atransient Agrobacterium tumefaciens-based delivery system in intact tissues.It was shown that the interferenceeffect was completely blocked by co-infiltration with HC-Pro plus siRNA constructs in both systemic andhypersensitive hosts.In the system host,all plants agro-infiltrated with HC-Pro plus siRNA constructsdisplayed the same symptoms as the negative control.Meanwhile,TMV RNA accumulation was found to beabundant in the upper leaves using reverse transcriptase-PCR (RT-PCR) and Northern blot assays.On thecontrary,plants agro-infiltrated with the siRNA construct alone were free of symptoms.Therefore,ourstudy suggests that the transient expression of HC-Pro inhibited the siRNA-directed host defenses againstTMV infection.

关键词： siRNA HC-Pro tobacco mosaic virus suppressor

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Transiently Expressed Short Hairpin RNA Targeting 126 kDa Protein of tobacco mosaic virus Interferes with virus Infection

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Acta Biochimica et Biophysica Sinica 2006年第1期38卷 22-28页

作者： Ming-Min ZHAO~(1,2),De-Rong AN~(1*),Jian ZHAO~2,Guang-Hua HUANG~3,Zu-Hua HE~4,and Jiang-Ye CHEN~3 ~1 College of Plant Protection,Northwest Science and Technology University of Agriculture and Forestry,Yangling 712100,China ~2 College of Agriculture,Yangtze University,Jingzhou 434025,China ~3 State Key Laboratory of Molecular Biology,Institute of Biochemistry and Cell Biology,Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences,Shanghai 200031,China ~4 State Key Laboratory of Plant Molecular Genetics,Institute of Plant Physiology and Ecology,Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences,Shanghai 200032,China College of Plant Protection Northwest Science and Technology University of Agriculture and Forestry Yang ling 712100 China College of Agriculture Yangtze University Jingzhou 434025 China State Key Laboratory of Molecular Biology Institute of Biochemistry and Cell Biology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200031 China State Key Laboratory of Plant Molecular Genetics Institute of Plant Physiology and Ecology Shanghai Institutes for Biological Sciences Chinese Academy of Sciences Shanghai 200032 China

RNA-interference (RNAi) silences gene expression by'guiding mRNA degradation in asequence-specific fashion.Small interfering RNA (siRNA),an intermediate of the RNAi pathway,has beenshown to be very effective in inhibiting virus infection in mammalian cells and cultured plant cells.Here,wereport that Agrobacterium tumefaciens-mediated transient expression of short hairpin RNA (shRNA) couldinhibit tobacco mosaic virus (TMV) RNA accumulation by targeting the gene encoding the replication-asso-ciated 126 kDa protein in intact plant tissue.Our results indicate that transiently expressed shRNA efficientlyinterfered with TMV infection.The interference observed is sequence-specific,and time-and site-dependent.Transiently expressed shRNA corresponding to the TMV 126 kDa protein gene did not inhibit cucumbermosaic virus (CMV),an unrelated tobamovirus.In order to interfere with TMV accumulation in tobaccoleaves,it is essential for the shRNA constructs to be infiltrated into the same leaves as TMV inoculation.Ourresults support the view that RNAi opens the door for novel therapeutic procedures against virus diseases.We propose that a combination of the RNAi technique and Agrobacterium-mediated transient expressioncould be employed as a potent antiviral treatment in plants.

关键词： tobacco mosaic virus 126 kDa protein RNA interference short hairpin RNA

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A bacterial protein Rhp-PSP inhibits plant viral proliferation through endoribonuclease activity

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Journal of Integrative Agriculture 2024年第6期23卷 1967-1978页

作者： Lijie Chen Qianze Peng Xiaohua Du Weixing Zhang Ju’e Cheng Shu’e Sun Deyong Zhang Pin Su Yong Liu Longping Branch College of BiologyHunan UniversityChangsha 410082China State Key Laboratory of Hybrid Rice/Institute of Plant Protection Hunan Academy of Agricultural SciencesChangsha 410125China

Plant virus causes massive crop losses globally.However,there is currently no effective measure to control plant viral disease.Previously,we identify an antiviral protein Rhp-PSP,produced by the bacterial Rhodopseudomonas palustris strain JSC-3b.In this study,we discover that the antiviral activity of Rhp-PSP relies on its endoribonuclease activity.Converting the arginine(R)residue at position 129 onto alanine(A)abolishs its endoribonuclease activity on coat protein(CP)RNA of tobacco mosaic virus(TMV),consequentially,compromises the antiviral activity of Rhp-PSP.Further investigation demonstrates that,the mutant Rhp-PSP^(R129A)is unable to form the homotrimer as the wild type,indicating the importance of quaternary junction for the endoribonuclease activity.Overexpression of Rhp-PSP in Nicotiana benthamiana significantly enhances the resistance against TMV of seedlings,while expression of Rhp-PSP^(R129A)did not,confirming that endoribonuclease activity is responsible for the antiviral activity of Rhp-PSP.In addition,foliar spray of Rhp-PSP solution on tomato and pepper plants significantly reduces the disease index of viral diseases,indicating that Rhp-PSP shows potential to develop antiviral agent in practice.

关键词： Rhodopseudomonas palustris Rhp-PSP tobacco mosaic virus endoribonuclease antiviral activity

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Hyaluronic acid-based hydrogels with tobacco mosaic virus containing cell adhesive peptide induce bone repair in normal and osteoporotic rats

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Biomaterials Translational 2020年第1期1卷 89-98页

作者： Jishan Yuan Panita Maturavongsadit Zhihui Zhou Bin Lv Yuan Lin Jia Yang Jittima Amie Luckanagul Department of Orthopaedic Surgery the Affiliated First People’s Hospital to Jiangsu UniversityZhenjiangJiangsu ProvinceChina Department of Chemistry and Biochemistry University of South CarolinaColumbiaSCUSA Joint Department of Biomedical Engineering North Carolina State University and The University of North Carolina at Chapel HillChapel HillNCUSA The State Key Laboratory of Polymer Physics and Chemistry Changchun Institute of Applied ChemistryChinese Academy of SciencesChangchunJilin ProvinceChina Department of Pharmaceutics and Industrial Pharmacy Faculty of Pharmaceutical SciencesChulalongkorn UniversityBangkokThailand Research Unit for Plant-produced Pharmaceuticals Chulalongkorn UniversityBangkokThailand.

tobacco mosaic virus(TMV)has been studied as a multi-functional agent for bone tissue engineering.An osteo-inductive effect of wild-type TMV has been reported,as it can significantly enhance the bone differentiation potential of bone marrow stromal cells both on a two-dimensional substrate and in a three-dimensional(3D)hydrogel system.A TMV mutant(TMV-RGD1)was created which featured the adhesion peptide arginyl-glycyl-aspartic acid(RGD),the most common peptide motif responsible for cell adhesion to the extracellular matrix,on the surface of the virus particle to enhance the bio-functionality of the scaffold material.We hypothesised that the incorporation of either wild-type TMV or TMV-RGD1 in the 3D hydrogel scaffold would induce bone healing in critical size defects of the cranial segmental bone.We have previously tested the virus-functionalised scaffolds,in vitro,with a hyaluronic acid-based system as an in-situ hydrogel platform for 3D cell encapsulation,culture,and differentiation.The results of these experiments suggested the potential of the virus-functionalised hydrogel to promote in vitro stem cell differentiation.The hydrogel-forming system we employed was shown to be safe and biocompatible in vivo.Here,we further explored the physiological responses regarding bone regeneration of a calvarial defect in both normal and osteoporotic ovariectomized rat models.Our results,based on histological analysis in both animal models,suggested that both wild-type TMV and TMV-RGD1 functionalised hydrogels could accelerate bone regeneration,without systemic toxicity,evaluated by blood counts.New bone formation was intensified by the incorporation of the RGD-mutant viral particles.This finding increased the potential for use of the rodshaped plant virus as a platform for the addition of powerful biofunctionality for tissue engineering applications.This study was approved by the Ethics Committee on Animal Use of the Zhenjiang Affiliated First People’s Hospital affiliated to Jiangsu University.

关键词： bone regeneration calvarial defect hydrogel osteoporosis tobacco mosaic virus

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Analysis of Spatial Distribution Pattern of tobacco mosaic virus (TMV) Based on Geostatistics

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Plant Diseases and Pests 2012年第1期3卷 62-63,68页

作者： WEI Guo-sheng BAI Sheng HUANG Dian LIE Fan-jun XIA Peng-liang Hubel Enshi tobacco Company Enshi 445600China

Using geostatistical method, the semi-variation function models of tobacco mosaic virus （TMV） in east-west and north-south directions were established, and the distribution pattern of TMV in large scale space was studied. The results showed that the distribution pattern of the disease in east-west and north-south directions belonged to linear model with abutment, and the spatial distribution pattern within the investigated areas was aggregated model. The spatial correlation distances in east-west and north-south directions were 29.953 4 and 47.813 8 km, and the spatial variabilities were 95.71% and 80.05%, respectively. This indicated that they had strong spatial correlation. Isoline map accessed by Kringing interpolation method could clearly reflect the spatial aggregated model.

关键词： Geostatistics tobacco mosaic virus Isoline map China

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Coat protein gene and 3' non-coding region of tobacco mosaic virus and tomato mosaic virus are associated with viral pathogenesis in Nicotiana tabacum

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Progress in Natural Science:Materials International 2002年第9期 41-45页

作者： XUE Chaoyang, TAO Xiaorong and ZHOU Xueping(Institute of Biotechnology, Zhejiang University, Hangzhou 310029, China) Institute of Biotechnology Zhejiang University Hangzhou 310029 China

The camellia isolate of tomato mosaic virus (ToMV-TL) can induce local necrotic lesions on the inoculated leaves in Nicotiana tabacum, whereas the broad bean isolate of tobacco mosaic virus (TMV-B) produces the mosaic symptom on systemic leaves. To examine viral determinant for differential infection phenotype in N. tabacum, the coat protein gene and the 3’ non-coding region of TMV was replaced with that of ToMV, the chimeric virus induced similar local necrotic lesions to that induced by ToMV. The results indicate that the coat protein gene and the 3’ non-coding region of TMV and ToMV influence the virus-induced pathogenesis in N. tabacum .

关键词： tobacco mosaic virus tomato mosaic virus coat protein 3’ non-coding region pathogenesis Nicotiana tabacum

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tobacco Alpha-expansin EXPA4 Involves in Nicotiana benthamiana Defense Against tobacco mosaic virus

Tobacco Alpha-expansin EXPA4 Involves in Nicotiana benthamia...

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中国植物病理学会2017年学术年会

作者： Chen Lijuan Zou Wenshan Wu Guo Lin Honghui Xi Dehui Ministry of Education Key Laboratory for Bio-Resource and Eco-Environment College of Life ScienceSichuan University

Expansins are cell-wall-loosening protein,known for their endogenous function in cell wall extensibility during plant growth.Expansin genes are classified into four subfamilies:or-and β-expansins（EXPA and EXPB,respectively）,expansin-like A（EXLA） and expansin-like B（EXLB）.Effects of expansins on plant growth,developmental processes and environment stress responses were well studied.By contrast,relatively limited information is available on expansins regulation of plant response to pathogen attack.Only several studies reported that plants response to pathogen may involve changes in the expression levels of expansin genes.For instance,Cantu et al.identified that simultaneous suppression of polygalacturonase（LePG） and LeExpl dramatically reduced the susceptibility of ripening fruit to necrotrophic fungi Botrytis cinerea.The absence or down-regulation of AtEXLA2 leads to increased resistance of Arabidopsis to B.cinerea in a CORONATINE INSENSITIVE 1（COIl）-dependent manner.By contrast,the roles of expansin genes in plant defense response to virus pathogen have not been studied in any detail.In this study,virus-induced gene silencing（VIGS） combined with agrobacterium-mediated transient overexpression method were conducted to investigate the role of Nicotiana tabacum alpha-expansin 4（EXPA4） in modulating tobacco mosaic virus（TMV-GFP） resistance in Nicotiana benthamiana.Results indicated that silencing of EXPA4 reduced the sensitivity of N.benthamiana to TMV-GFP,and overexpression of EXPA4 accelerated the virus reproduction on inoculated plants.In addition,our data suggested that the effects of EXPA4 on tobacco defense against virus attack were caused by affecting the antioxidative metabolism and phytohormone-mediated immunity responses in tobacco.The changes of EXPA4 expression has more effects on SA and ET not JA in N.benthamiana induced by TMV-GFP.The expressions of SA and ET-mediated defense genes were also significantly affected by EXPA4,indicating that the effects of EXPA4 on the susceptibility of N.benthamiana to TM

关键词： Expansins tobacco mosaic virus Nicotiana benthamiana Plant defense

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Bacillus atrophaeus strain HAB-5 promotes vegetative growth of tobacco and its secretion metabolites protect plants against tobacco mosaic virus infection

Bacillus atrophaeus strain HAB-5 promotes vegetative growth ...

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中国植物病理学会第十一届全国会员代表大会暨2018年学...

作者： Mamy Jayne Nelly Rajaofera Jin Pengfei Shen Haiyan Hafiz Hasnain Nawaz Wang Yi He Qiguang Xu Liangxiang Cui Hongguang Liu Wenbo Miao Weiguo Institute of Tropical Agriculture and Forestry Hainan UniversityHainan Key Laboratory for Sustainable Utilization of Tropical Bioresource

Finding alternatives of synthetic pesticide for health and the safe environment has become a crucial issue for scientific research.A number of studies have reported efficacy of Bacillus species on promoting plant development,as well as protecting plants against pathogens invasion,especially pathogenic fungi and bacteria.However,little was known about Bacillus species in controlling viral diseases.In this study,Bacillus atrophaeus strain HAB-5,isolated from soil of cotton field,Xinjiang,China,was able to efficiently promote the growth of tobacco plant in that the expansins genes NtEXPl and NtEXP2 in tobacco leaves after treatment with HAB-5.Then tobacco mosaic virus（TMV）/Nicotiana tobacco system was employed to evaluate virus resistance induced by HAB-5.tobacco leaves were treated with antimicrobial metabolites of HAB-5 strain（lmg/mL）,and 12 hours later the treated leaves were challenged with TMV via rub-inoculation.The results showed that diseasing symptoms were obviously compromised in tobacco leaves treated with HAB-5,and viral accumulation levels were also greatly reduced.Moreover,it was found that the signaling regulatory gene（NPR1）,defense genes（PR-1a,PR-1b,Chia5）,and hypertensive response related genes（Hsr203J,Hinl） were up-regulated in plants treated with the metabolites.Altogether,these accumulated results strongly supported the strain HAB-5 to be a biocontrol strain against TMV.

关键词： Bacillus atrophaeus tobacco mosaic virus Biocontrol

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Design,Synthesis and Biological Evaluation of Novel N-(4-([2,2':5',2''-Terthiophen]-5-yl)-2-methylbut-3-yn-2-yl)Benzamide Derivatives

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中国化学:英文版 2015年第2期33卷 268-276页

作者： Youquan Zhu Haili Zhang Danyang Wang Lei Li Han Xu Huabin Li Yuan Ma State Key Laboratory of Elemento-Organic Chemistry Institute of Elemento-Organic ChemistryCollaborative Innovation Center of Chemical Science and Engineering(Tianjin)Nankai UniversityTianjin 300071China

On the basis of the principle of combination of active groups,a series of novel N-(4-([2,2':5',2''-terthiophen]-5-yl)-2-methylbut-3-yn-2-yl)benzamide derivatives were designed,synthesized and systematically evaluated for their antiviral activity against tobacco mosaic virus(TMV).The bioassay results showed that most of these compounds displayed good anti-TMV activity,and some of them exhibited higher antiviral activity than commercial Ningnanmycin.Especially,compound 8e with excellent anti-TMV activity(inactivation activity,92.3%/500μg•mL^(−1);curative activity,85.7%/500μg•mL^(−1) and protection activity,64.7%/500μg•mL^(−1))emerged as a potential inhibitor of plant virus TMV.Quantitative structure-activity relationship studies proved that the van der Waals volume(V)and electronic parameter(Σ(Σσo+σp)andΣσm)for the substituent R1 were very important for antiviral activities in this class of compounds.

关键词： α-terthienyl benzamide antiviral activity tobacco mosaic virus tobacco mosaic virus(TMV)

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