DOCSLIB.ORG

Development of a Full-Length Infectious Clone of Grapevine Rupestris Stem Pitting-Associated Virus Strain Syrah and GFP-Tagged and VIGS Vectors for Vitis Vinifera

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Development of a Full-Length Infectious Clone of Grapevine Rupestris Stem Pitting-Associated Virus Strain Syrah and GFP-Tagged and VIGS Vectors for Vitis Vinifera Development of a Full-Length Infectious Clone of Grapevine rupestris stem pitting-associated virus strain Syrah and GFP-Tagged and VIGS Vectors for Vitis vinifera. by Olivia M. A. Roscow A thesis presented to The University of Guelph In partial fulfilment of requirements for the degree of Master of Science in Molecular and Cellular Biology Guelph, Ontario, Canada c Olivia M. A. Roscow, December, 2019 ABSTRACT Development of a Full-Length Infectious Clone of Grapevine rupestris stem pitting-associated virus strain Syrah and Production of GFP-Tagged and VIGS Vectors for Vitis vinifera. Olivia Marion Alice Roscow Advisor: University of Guelph, 2019 Dr. Baozhong Meng Advisory Committee: Dr. Annette Nassuth Dr. Ian Tetlow Grapevines are an important Canadian crop, yet quality, health, and yield are threatened by viral infections and coinfections with multiple viruses complicate disease associations. Grapevine rupestris stem pitting-associated virus (GRSPaV) is a single-stranded, positive-sense RNA virus that is widely distributed worldwide and is associated with Rupestris Stem Pitting, Syrah Decline, and Grapevine Vein Necrosis. Specifically the SY strain is associated with unhealthy vines, possibly being more pathogenic. An infectious clone can help clarify disease associa- tions and viral biology by reproducing singular infections in virus-free plants. MGT, a clone related to SY, was constructed and a GFP-tagged version produced GFP faster and to a greater degree than the asymptomatic GG strain in Nicotiana benthamiana, as determined by Western blotting and epifluorescence microscopy. A Virus-Induced Gene Silencing (VIGS) clone was also constructed for future study of grapevine genomics. These N. benthamiana experiments provide a foundation for continued work with this clone with grapevines. ACKNOWLEDGEMENTS Thanks to Dr. Baozhong Meng for instilling in me a passion for research and for challenging me to overcome seemingly insurmountable obstacles. He has not only nurtured my interest in all things molecular and inspired me to further pursue research, but also taught me great life lessons on how to be stoic and that no challenge is unconquerable. Thanks to Sunny Li for being a great friend and colleague. She provided me with new ideas in times of need and was a great companion in the lab. I hope to always remain in contact. Thanks to everyone who has passed through and is currently working in the Meng lab. Special thanks to Clayton Moore for helping me adjust to Ontario, Huogen Xiao for helping me adjust to the lab, and Mehdi Shabanian for his patience and guidance. Thanks to my advisory committee, Annette Nassuth and Ian Tetlow, for their enthusi- asm and suggestions. They both were tremendously helpful and indispensable in my studies. Thanks to Rosemarie Ganassin, Mark Polinski, Kyler Garver, and Caroline Josefsson for putting me on the right track. Without their help, I would not have progressed this far. Thanks to my immediate family (Greg, Alison, Frazer, and Edward), my partner War- wick, and my close friends for supporting me through the highs and lows of research. No path is linear and they accompanied me through the winding turns. Last, but not least, thanks to NSERC, for their funding and financial support, without which I could not have pursued my passion. Special thanks to the University of Guelph for awarding me the Graduate Excellence Entrance Scholarship and to the Province of Ontario for awarding me the Queen Elizabeth II Graduate Scholarship in Science & Technology. iii TABLE OF CONTENTS ABSTRACT ...................................... ii TABLE OF CONTENTS ............................... vii LIST OF TABLES ................................... viii LIST OF FIGURES .................................. x LIST OF ABBREVIATIONS ............................. xi 1 INTRODUCTION ................................ 1 1.1 Grapevines . 1 1.1.1 History . 1 1.1.2 Viruses . 2 1.1.3 Defence & Immunity Against Viruses . 3 1.2 Grapevine rupestris stem pitting-associated virus ............ 4 1.2.1 Distribution & Transmission . 5 1.2.2 Taxonomy & Phylogeny . 5 1.2.3 Genome . 6 1.2.4 Replication & Life Cycle . 10 1.2.5 Disease Associations . 12 1.3 Viral Vectors . 13 1.3.1 Wildtype Clones . 15 1.3.2 Protein Expression Vectors . 16 1.3.3 Virus-Induced Gene Silencing (VIGS) Vectors . 17 1.3.4 GRSPaV-GG Clone . 18 1.4 Justification & Goals . 19 1.5 Hypotheses . 20 1.6 Objectives . 20 2 MATERIALS & METHODS .......................... 20 2.1 Full Length Clone . 20 iv 2.1.1 Previous Work . 22 2.1.2 Viral Source & Cloning Genome Fragments . 22 2.1.3 Subcloning Fragments D and A into the Intermediate Vector . 23 2.1.4 Subcloning the Terminator into the Intermediate Vector . 24 2.1.5 Removing Restriction Sites . 24 2.1.6 Subcloning Fragments B1, B2 & C into the Intermediate Vector 24 2.2 Wildtype Clone . 26 2.3 VIGS Clone . 27 2.3.1 Inserting Restriction Sites into Fragment D . 27 2.3.2 PDS Cloning . 29 2.3.3 Subcloning PDS into Fragment D . 29 2.3.4 Cloning SGP into Fragment D . 29 2.3.5 Reinserting Missing Viral Sequence . 30 2.3.6 Subcloning recombinant PDS Fragment into Full Length Clone and Binary Vector . 30 2.3.7 Subcloning PDS into GRSPaV-GG . 30 2.4 GFP Expression Vector . 31 2.4.1 GFP Cloning . 31 2.4.2 Reinserting Missing Viral Sequence . 31 2.4.3 Subcloning recombinant GFP Fragment into Full Length Clone and Binary Vector . 32 2.4.4 Subcloning GFP into GRSPaV-GG . 32 2.5 Infectivity Assays in N. benthamiana ................... 32 2.6 Infectivity Assays in V. vinifera ...................... 32 2.7 Analyses . 33 2.7.1 Genome Sequencing . 33 2.7.2 Symptom observation . 33 2.7.3 Western Blotting . 33 2.7.4 Fluorescence Microscopy . 35 v 2.7.5 Bioinformatic & Statistical Analyses . 35 3 RESULTS ..................................... 36 3.1 Construction of wildtype and GFP-tagged full-length clones for GRSPaV isolate VD-102 . 36 3.2 Infectivity of wildtype and GFP-tagged MGT clones in N. benthamiana .............................. 38 3.3 RNA & Protein Structure Predictions . 39 3.4 Effectiveness of pRSPMGT-PDSf and pRSPGG-PDSf as VIGS vectors in N. benthamiana ............................ 40 4 DISCUSSION .................................. 55 5 CONCLUSION .................................. 60 REFERENCES ..................................... 61 A APPENDIX .................................... 78 A.1 Standard Procedures and Procedures . 78 A.1.1 Agarose Gel Purification of DNA . 78 A.1.2 Agrobacterium Transformation . 78 A.1.3 Agroinfiltration of N. benthamiana and V. vinifera ...... 78 A.1.4 Broth Culture and Plasmid Miniprep . 79 A.1.5 Competent A. tumefaciens Preparation . 79 A.1.6 Competent E. coli Preparation . 79 A.1.7 Dephosphorylation . 80 A.1.8 Digestion . 80 A.1.9 E. coli Transformation . 80 A.1.10 KOD Polymerase Chain Reaction (PCR) . 81 A.1.11 Ligation . 81 A.1.12 Overlap-Extension PCR (OE-PCR) . 81 A.1.13 PaCeR PCR . 82 A.1.14 RNA Extraction . 82 A.1.15 Reverse Transcription (RT) . 82 vi A.1.16 Site-Directed Mutagenesis (SDM) . 82 A.1.17 TA cloning . 83 A.1.18 Taq PCR/Colony PCR . 83 A.2 MGT Consensus Sequence . 83 A.3 Supplementary Figures and Tables . 84 vii LIST OF TABLES 1: Percent nucleotide identity between TA cloned fragments and the GRSPaV-SY and VD-102 genomes. 37 A1: Percent similarity (bottom left) and hamming distance (top right) for genomes/proteins of prominent GRSPaV strains. 90 A2: Primers used for cloning and diagnostic assays. 93 viii LIST OF FIGURES 1: Representative diagram of the GRSPaV genome. 6 2: Syrah Decline (SD) symptoms. 13 3: Rugose Wood Complex (RWC) symptoms. 14 4: Grapevine vein Necrosis (GVN) symptoms in the indicator rootstock 110R. 15 5: Cloning plan based on VD-102 (MF979534.1) isolate. 21 6: Vector map for pCB-301.3. 26 7: Plan for creating GFP-tagged and VIGS vectors from the wildtype GRSPaV- SY clone. 28 8: Coverage Diagram of plan to sequence entire wildtype GRSPaV-SY clone (pBS-SY) genome. 34 9: Multiple sequence alignment of the pRSPMGT and 13 prominent GRSPaV strains shows a gap in VD-102 sequencing. 38 10: Phylogenetic tree of all GRSPaV isolates for which complete or near complete genomes are available. 41 11: Western blot for detecting GRSPaV CP from N. benthamiana agroinfiltrated with pRSPMGT-GFP, pRSPGG-GFP, and pCB-301.3 (–) at 10 days post-infiltration (dpi). 42 12: N. benthamiana infiltrated with pRSPMGT-GFP from 2 to 10 days post-infiltration (dpi) at 100x magnification. 43 13: N. benthamiana infiltrated with pRSPGG-GFP from 2 to 10 days post-infiltration (dpi) at 100x magnification. 44 14: N. benthamiana infiltrated with pRSPGG-GFP (left) and pRSPMGT-GFP (right) in the same leaf from 2 to 10 days post-infiltration (dpi) at 100x magnification. 45 15: Western blot of pooled protein extracts (n=3) for detecting GFP from N. ben- thamiana agroinfiltrated with pRSPMGT-GFP, pRSPGG-GFP, and pCB-301.3 (–) at 2, 3, 5, 7, and 10 days post-infiltration (dpi). 46 16: Analysis of GFP bands from a Western blot for N. benthamiana agroinfiltrated with pRSPMGT-GFP and pRSPGG-GFP from 2 to 10 days post-infiltration (dpi). A: Normalized density values. B: Normalized percent differences in density values relative to 2 dpi. 47 17: Predicted 2◦ and 3◦ RNA structures of the (+)-sense strands of the 3’ UTRs of MGT and GG. 48 18: Predicted 3◦ protein structures of the replicase domains of MGT (red) and GG (grey). 49 19: Predicted 3◦ protein structures of the proteins of MGT (red) and GG (grey). 50 20: Hydrophobicity plots of proteins for MGT (black) and GG (red). 51 21: Predicted 2◦ and 3◦ RNA structures of the (–)-sense strands of the 97 nt CP SGP region of MGT and GG. 52 22: Predicted 2◦ and 3◦ RNA structures of the (–)-sense strands of the 70 nt CP SGP region minus the CP sgRNA 5’ UTR of MGT and GG.
Load more

Recommended publications
  • Grapevine Virus Diseases: Economic Impact and Current Advances in Viral Prospection and Management1
    1/22 ISSN 0100-2945 http://dx.doi.org/10.1590/0100-29452017411 GRAPEVINE VIRUS DISEASES: ECONOMIC IMPACT AND CURRENT ADVANCES IN VIRAL PROSPECTION AND MANAGEMENT1 MARCOS FERNANDO BASSO2, THOR VINÍCIUS MArtins FAJARDO3, PASQUALE SALDARELLI4 ABSTRACT-Grapevine (Vitis spp.) is a major vegetative propagated fruit crop with high socioeconomic importance worldwide. It is susceptible to several graft-transmitted agents that cause several diseases and substantial crop losses, reducing fruit quality and plant vigor, and shorten the longevity of vines. The vegetative propagation and frequent exchanges of propagative material among countries contribute to spread these pathogens, favoring the emergence of complex diseases. Its perennial life cycle further accelerates the mixing and introduction of several viral agents into a single plant. Currently, approximately 65 viruses belonging to different families have been reported infecting grapevines, but not all cause economically relevant diseases. The grapevine leafroll, rugose wood complex, leaf degeneration and fleck diseases are the four main disorders having worldwide economic importance. In addition, new viral species and strains have been identified and associated with economically important constraints to grape production. In Brazilian vineyards, eighteen viruses, three viroids and two virus-like diseases had already their occurrence reported and were molecularly characterized. Here, we review the current knowledge of these viruses, report advances in their diagnosis and prospection of new species, and give indications about the management of the associated grapevine diseases. Index terms: Vegetative propagation, plant viruses, crop losses, berry quality, next-generation sequencing. VIROSES EM VIDEIRAS: IMPACTO ECONÔMICO E RECENTES AVANÇOS NA PROSPECÇÃO DE VÍRUS E MANEJO DAS DOENÇAS DE ORIGEM VIRAL RESUMO-A videira (Vitis spp.) é propagada vegetativamente e considerada uma das principais culturas frutíferas por sua importância socioeconômica mundial.
    [Show full text]
  • Recent Advances on Detection and Characterization of Fruit Tree Viruses Using High-Throughput Sequencing Technologies
    viruses Review Recent Advances on Detection and Characterization of Fruit Tree Viruses Using High-Throughput Sequencing Technologies Varvara I. Maliogka 1,* ID , Angelantonio Minafra 2 ID , Pasquale Saldarelli 2, Ana B. Ruiz-García 3, Miroslav Glasa 4 ID , Nikolaos Katis 1 and Antonio Olmos 3 ID 1 Laboratory of Plant Pathology, School of Agriculture, Faculty of Agriculture, Forestry and Natural Environment, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece; [email protected] 2 Istituto per la Protezione Sostenibile delle Piante, Consiglio Nazionale delle Ricerche, Via G. Amendola 122/D, 70126 Bari, Italy; [email protected] (A.M.); [email protected] (P.S.) 3 Centro de Protección Vegetal y Biotecnología, Instituto Valenciano de Investigaciones Agrarias (IVIA), Ctra. Moncada-Náquera km 4.5, 46113 Moncada, Valencia, Spain; [email protected] (A.B.R.-G.); [email protected] (A.O.) 4 Institute of Virology, Biomedical Research Centre, Slovak Academy of Sciences, Dúbravská cesta 9, 84505 Bratislava, Slovak Republic; [email protected] * Correspondence: [email protected]; Tel.: +30-2310-998716 Received: 23 July 2018; Accepted: 13 August 2018; Published: 17 August 2018 Abstract: Perennial crops, such as fruit trees, are infected by many viruses, which are transmitted through vegetative propagation and grafting of infected plant material. Some of these pathogens cause severe crop losses and often reduce the productive life of the orchards. Detection and characterization of these agents in fruit trees is challenging, however, during the last years, the wide application of high-throughput sequencing (HTS) technologies has significantly facilitated this task. In this review, we present recent advances in the discovery, detection, and characterization of fruit tree viruses and virus-like agents accomplished by HTS approaches.
    [Show full text]
  • 2008.003P (To Be Completed by ICTV Officers)
    Taxonomic proposal to the ICTV Executive Committee This form should be used for all taxonomic proposals. Please complete all those modules that are applicable (and then delete the unwanted sections). Code(s) assigned: 2008.003P (to be completed by ICTV officers) Short title: 1 new species in the genus Foveavirus (e.g. 6 new species in the genus Zetavirus; re-classification of the family Zetaviridae etc.) Modules attached 1 2 3 4 5 (please check all that apply): 6 7 Author(s) with e-mail address(es) of the proposer: Mike Adams ([email protected]) on behalf of the Flexiviridae SG ICTV-EC or Study Group comments and response of the proposer: MODULE 5: NEW SPECIES Code 2008.003P (assigned by ICTV officers) To create 1 new species assigned as follows: Fill in all that apply. Ideally, species Genus: Foveavirus should be placed within a genus, but it is Subfamily: acceptable to propose a species that is Family: proposed family Betaflexiviridae within a Subfamily or Family but not (formerly Flexiviridae) assigned to an existing genus (in which case put “unassigned” in the genus box) Order: Name(s) of proposed new species: Peach chlorotic mottle virus Argument to justify the creation of the new species: If the species are to be assigned to an existing genus, list the criteria for species demarcation and explain how the proposed members meet these criteria. Species demarcation criteria published in the 8th report are: Each distinct species usually has a specific natural host range. Distinct species do not cross-protect in infected common host plant species.
    [Show full text]
  • Quality Assessment and Validation of High-Throughput Sequencing for Grapevine Virus Diagnostics
    viruses Article Quality Assessment and Validation of High-Throughput Sequencing for Grapevine Virus Diagnostics Nourolah Soltani 1,† , Kristian A. Stevens 2,3,4,†, Vicki Klaassen 2, Min-Sook Hwang 2, Deborah A. Golino 1 and Maher Al Rwahnih 1,* 1 Department of Plant Pathology, University of California-Davis, Davis, CA 95616, USA; [email protected] (N.S.); [email protected] (D.A.G.) 2 Foundation Plant Services, University of California-Davis, Davis, CA 95616, USA; [email protected] (K.A.S.); [email protected] (V.K.); [email protected] (M.-S.H.) 3 Department of Computer Science, University of California-Davis, Davis, CA 95616, USA 4 Department of Evolution and Ecology, University of California-Davis, Davis, CA 95616, USA * Correspondence: [email protected] † These authors contributed equally to this work. Abstract: Development of High-Throughput Sequencing (HTS), also known as next generation sequencing, revolutionized diagnostic research of plant viruses. HTS outperforms bioassays and molecular diagnostic assays that are used to screen domestic and quarantine grapevine materials in data throughput, cost, scalability, and detection of novel and highly variant virus species. However, before HTS-based assays can be routinely used for plant virus diagnostics, performance specifications need to be developed and assessed. In this study, we selected 18 virus-infected grapevines as a test panel for measuring performance characteristics of an HTS-based diagnostic assay. Total nucleic acid (TNA) was extracted from petioles and dormant canes of individual samples and constructed Citation: Soltani, N.; Stevens, K.A.; libraries were run on Illumina NextSeq 500 instrument using a 75-bp single-end read platform.
    [Show full text]
  • Small Hydrophobic Viral Proteins Involved in Intercellular Movement of Diverse Plant Virus Genomes Sergey Y
    AIMS Microbiology, 6(3): 305–329. DOI: 10.3934/microbiol.2020019 Received: 23 July 2020 Accepted: 13 September 2020 Published: 21 September 2020 http://www.aimspress.com/journal/microbiology Review Small hydrophobic viral proteins involved in intercellular movement of diverse plant virus genomes Sergey Y. Morozov1,2,* and Andrey G. Solovyev1,2,3 1 A. N. Belozersky Institute of Physico-Chemical Biology, Moscow State University, Moscow, Russia 2 Department of Virology, Biological Faculty, Moscow State University, Moscow, Russia 3 Institute of Molecular Medicine, Sechenov First Moscow State Medical University, Moscow, Russia * Correspondence: E-mail: [email protected]; Tel: +74959393198. Abstract: Most plant viruses code for movement proteins (MPs) targeting plasmodesmata to enable cell-to-cell and systemic spread in infected plants. Small membrane-embedded MPs have been first identified in two viral transport gene modules, triple gene block (TGB) coding for an RNA-binding helicase TGB1 and two small hydrophobic proteins TGB2 and TGB3 and double gene block (DGB) encoding two small polypeptides representing an RNA-binding protein and a membrane protein. These findings indicated that movement gene modules composed of two or more cistrons may encode the nucleic acid-binding protein and at least one membrane-bound movement protein. The same rule was revealed for small DNA-containing plant viruses, namely, viruses belonging to genus Mastrevirus (family Geminiviridae) and the family Nanoviridae. In multi-component transport modules the nucleic acid-binding MP can be viral capsid protein(s), as in RNA-containing viruses of the families Closteroviridae and Potyviridae. However, membrane proteins are always found among MPs of these multicomponent viral transport systems.
    [Show full text]
  • Grapevine Fanleaf Virus: Biology, Biotechnology and Resistance
    GRAPEVINE FANLEAF VIRUS: BIOLOGY, BIOTECHNOLOGY AND RESISTANCE A Dissertation Presented to the Faculty of the Graduate School of Cornell University In Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy by John Wesley Gottula May 2014 © 2014 John Wesley Gottula GRAPEVINE FANLEAF VIRUS: BIOLOGY, BIOTECHNOLOGY AND RESISTANCE John Wesley Gottula, Ph. D. Cornell University 2014 Grapevine fanleaf virus (GFLV) causes fanleaf degeneration of grapevines. GFLV is present in most grape growing regions and has a bipartite RNA genome. The three goals of this research were to (1) advance our understanding of GFLV biology through studies on its satellite RNA, (2) engineer GFLV into a viral vector for grapevine functional genomics, and (3) discover a source of resistance to GFLV. This author addressed GFLV biology by studying the least understood aspect of GFLV: its satellite RNA. This author sequenced a new GFLV satellite RNA variant and compared it with other satellite RNA sequences. Forensic tracking of the satellite RNA revealed that it originated from an ancestral nepovirus and was likely introduced from Europe into North America. Greenhouse experiments showed that the GFLV satellite RNA has commensal relationship with its helper virus on a herbaceous host. This author engineered GFLV into a biotechnology tool by cloning infectious GFLV genomic cDNAs into binary vectors, with or without further modifications, and using Agrobacterium tumefaciens delivery to infect Nicotiana benthamiana. Tagging GFLV with fluorescent proteins allowed tracking of the virus within N. benthamiana and Chenopodium quinoa tissues, and imbuing GFLV with partial plant gene sequences proved the concept that endogenous plant genes can be knocked down.
    [Show full text]
  • Ancient Recombination Events and the Origins of Hepatitis E Virus Andrew G
    Kelly et al. BMC Evolutionary Biology (2016) 16:210 DOI 10.1186/s12862-016-0785-y RESEARCH ARTICLE Open Access Ancient recombination events and the origins of hepatitis E virus Andrew G. Kelly, Natalie E. Netzler and Peter A. White* Abstract Background: Hepatitis E virus (HEV) is an enteric, single-stranded, positive sense RNA virus and a significant etiological agent of hepatitis, causing sporadic infections and outbreaks globally. Tracing the evolutionary ancestry of HEV has proved difficult since its identification in 1992, it has been reclassified several times, and confusion remains surrounding its origins and ancestry. Results: To reveal close protein relatives of the Hepeviridae family, similarity searching of the GenBank database was carried out using a complete Orthohepevirus A, HEV genotype I (GI) ORF1 protein sequence and individual proteins. The closest non-Hepeviridae homologues to the HEV ORF1 encoded polyprotein were found to be those from the lepidopteran-infecting Alphatetraviridae family members. A consistent relationship to this was found using a phylogenetic approach; the Hepeviridae RdRp clustered with those of the Alphatetraviridae and Benyviridae families. This puts the Hepeviridae ORF1 region within the “Alpha-like” super-group of viruses. In marked contrast, the HEV GI capsid was found to be most closely related to the chicken astrovirus capsid, with phylogenetic trees clustering the Hepeviridae capsid together with those from the Astroviridae family, and surprisingly within the “Picorna-like” supergroup. These results indicate an ancient recombination event has occurred at the junction of the non-structural and structure encoding regions, which led to the emergence of the entire Hepeviridae family.
    [Show full text]
  • A Diverse Virome of Leafroll-Infected Grapevine Unveiled by Dsrna Sequencing
    viruses Article A Diverse Virome of Leafroll-Infected Grapevine Unveiled by dsRNA Sequencing Mamadou L. Fall 1,* , Dong Xu 1, Pierre Lemoyne 1, Issam E. Ben Moussa 1,2, Carole Beaulieu 2 and Odile Carisse 1 1 Saint-Jean-sur-Richelieu Research and Development Centre, Agriculture and Agri-Food Canada, St-Jean-sur-Richelieu, Quebec, QC J3B 3E6, Canada; [email protected] (D.X.); [email protected] (P.L.); [email protected] (I.E.B.M.); [email protected] (O.C.) 2 Département de Biologie, Université de Sherbrooke, Sherbrooke, QC J1K 2R1, Canada; [email protected] * Correspondence: [email protected]; Tel.: +1-579-224-3024 Received: 28 August 2020; Accepted: 6 October 2020; Published: 8 October 2020 Abstract: Quebec is the third-largest wine grape producing province in Canada, and the industry is constantly expanding. Traditionally, 90% of the grapevine cultivars grown in Quebec were winter hardy and largely dominated by interspecific hybrid Vitis sp. cultivars. Over the years, the winter protection techniques adopted by growers and climate changes have offered an opportunity to establish V. vinifera L. cultivars (e.g., Pinot noir). We characterized the virome of leafroll-infected interspecific hybrid cultivar and compared it to the virome of V.vinifera cultivar to support and facilitate the transition of the industry. A dsRNA sequencing method was used to sequence symptomatic and asymptomatic grapevine leaves of different cultivars. The results suggested a complex virome in terms of composition, abundance, richness, and phylogenetic diversity. Three viruses, grapevine Rupestris stem pitting-associated virus, grapevine leafroll-associated virus (GLRaV) 3 and 2 and hop stunt viroid (HSVd) largely dominated the virome.
    [Show full text]
  • Evidence to Support Safe Return to Clinical Practice by Oral Health Professionals in Canada During the COVID-19 Pandemic: a Repo
    Evidence to support safe return to clinical practice by oral health professionals in Canada during the COVID-19 pandemic: A report prepared for the Office of the Chief Dental Officer of Canada. November 2020 update This evidence synthesis was prepared for the Office of the Chief Dental Officer, based on a comprehensive review under contract by the following: Paul Allison, Faculty of Dentistry, McGill University Raphael Freitas de Souza, Faculty of Dentistry, McGill University Lilian Aboud, Faculty of Dentistry, McGill University Martin Morris, Library, McGill University November 30th, 2020 1 Contents Page Introduction 3 Project goal and specific objectives 3 Methods used to identify and include relevant literature 4 Report structure 5 Summary of update report 5 Report results a) Which patients are at greater risk of the consequences of COVID-19 and so 7 consideration should be given to delaying elective in-person oral health care? b) What are the signs and symptoms of COVID-19 that oral health professionals 9 should screen for prior to providing in-person health care? c) What evidence exists to support patient scheduling, waiting and other non- treatment management measures for in-person oral health care? 10 d) What evidence exists to support the use of various forms of personal protective equipment (PPE) while providing in-person oral health care? 13 e) What evidence exists to support the decontamination and re-use of PPE? 15 f) What evidence exists concerning the provision of aerosol-generating 16 procedures (AGP) as part of in-person
    [Show full text]
  • Analyses of Virus/Viroid Communities in Nectarine Trees by Next
    www.nature.com/scientificreports OPEN Analyses of virus/viroid communities in nectarine trees by next-generation sequencing Received: 4 February 2019 Accepted: 9 August 2019 and insight into viral synergisms Published: xx xx xxxx implication in host disease symptoms Yunxiao Xu1, Shifang Li1,2, Chengyong Na3, Lijuan Yang1 & Meiguang Lu 1 We analyzed virus and viroid communities in fve individual trees of two nectarine cultivars with diferent disease phenotypes using next-generation sequencing technology. Diferent viral communities were found in diferent cultivars and individual trees. A total of eight viruses and one viroid in fve families were identifed in a single tree. To our knowledge, this is the frst report showing that the most-frequently identifed viral and viroid species co-infect a single individual peach tree, and is also the frst report of peach virus D infecting Prunus in China. Combining analyses of genetic variation and sRNA data for co-infecting viruses/viroid in individual trees revealed for the frst time that viral synergisms involving a few virus genera in the Betafexiviridae, Closteroviridae, and Luteoviridae families play a role in determining disease symptoms. Evolutionary analysis of one of the most dominant peach pathogens, peach latent mosaic viroid (PLMVd), shows that the PLMVd sequences recovered from symptomatic and asymptomatic nectarine leaves did not all cluster together, and intra-isolate divergent sequence variants co-infected individual trees. Our study provides insight into the role that mixed viral/viroid communities infecting nectarine play in host symptom development, and will be important in further studies of epidemiological features of host-pathogen interactions. Peach is one of the most widely grown fruit crops in China, and nectarine (Prunus persica cv.
    [Show full text]
  • The Viroses and Virus-Like Diseases of the Grapevine
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by JKI Open Journal Systems (Julius Kühn-Institut) Vitis 25, 227-275 (1986) SCIENTIA VITIS ET VINI The viroses and virus-like diseases of the grapevine A bibliographic report, 1979-1984 by R. BOVEY1) and G. P. MARTELLI 2) Foreword The compilation of 'bibliographic reports' on virus and virus-like diseases of Vitis species was initiated in 1965, under the auspices of the International Council for the Study of Viruses and Virus Diseases of the Grapevine (ICVG). Three reports have already been published: CAUDWELL, A.; 1965: Bibliographie des viroses de la vigne des origines a 1965. Office International de la Vigne et du Vin, Paris, 76 pp. (references 1-1019). CAUDWELL, A.; HEWITT, W. B.; BOVEY, R.; 1972: Les viroses de Ja vigne. Bibliographie de 1965-1970. Vitis 11, 303-324 (references 1020-1386). HEWITT, W. B.; BOVEY, R.; 1979: The viroses and virus-like diseases of the grapevine. A bibliographic report 1971-1978. Vitis 18, 316-376 (references 1387-2163). The present report constitues the fourth of the series, covering the period from 1979 through 1984. lt includes 20 references (2164-2183) of papers published prior to 1979, which had been omitted in previous lists and all papers presented at the 8th Meeting of ICVG held in Septe mber 1984 in Bari, Italy, whose Proceedings were pub­ lished in the first 1985 issue of Phytopathologia Mediterranea. 636 i·eferences of research papers 01· reviews on virus, mycoplasma-like organisms and virus-like diseases of Vitis spp„ their causal agents, vectors, control meas­ ures and various aspects of practical applications of virological knowledge to the improvement of viticulture are contained in this presentation.
    [Show full text]
  • Disease Progression of Vector-Mediated Grapevine Leafroll-Associated Virus 3 Infection of Mature Plants Under Commercial Vineyard Conditions
    UC Berkeley UC Berkeley Previously Published Works Title Disease progression of vector-mediated Grapevine leafroll-associated virus 3 infection of mature plants under commercial vineyard conditions Permalink https://escholarship.org/uc/item/7dp9s67t Journal European Journal of Plant Pathology, 146(1) ISSN 0929-1873 Authors Blaisdell, GK Cooper, ML Kuhn, EJ et al. Publication Date 2016-09-01 DOI 10.1007/s10658-016-0896-8 Peer reviewed eScholarship.org Powered by the California Digital Library University of California Eur J Plant Pathol (2016) 146:105–116 DOI 10.1007/s10658-016-0896-8 Disease progression of vector-mediated Grapevine leafroll-associated virus 3 infection of mature plants under commercial vineyard conditions G. Kai Blaisdell & Monica L. Cooper & Emily J. Kuhn & Katey A. Taylor & Kent M. Daane & Rodrigo P. P. Almeida Accepted: 24 February 2016 /Published online: 2 March 2016 # Koninklijke Nederlandse Planteziektenkundige Vereniging 2016 Abstract Grapevine leafroll-associated virus 3 newly symptomatic vines in commercial vineyards (GLRaV-3) is associated with the economically damag- probably became infected during the previous growing ing grapevine leafroll disease, and is transmitted in a season, and a decline in berry quality can be expected semi-persistent manner by several mealybug species. during the same year in which symptoms appear. We performed the first controlled field study of vector- mediated inoculations with GLRaV-3 in a commercial Keywords Grapevine leafroll-associated virus-3 . vineyard with previously asymptomatic vines, and mon- Grapevine leafroll disease . Incubation time . itored the vines during four growing seasons. We then Pseudococcusmaritimus . Semi-persistent transmission . compared the outcome of vector-mediated inoculations Vitis vinifera in the field study to an analogous laboratory study.
    [Show full text]