DOCSLIB.ORG

Cauliflower Mosaic Virus Gene VI Produces a Symptomatic Phenotype in Transgenic Tobacco Plants (Plant DNA Virus/Virus Symptomology) GAIL A

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Cauliflower Mosaic Virus Gene VI Produces a Symptomatic Phenotype in Transgenic Tobacco Plants (Plant DNA Virus/Virus Symptomology) GAIL A Proc. Nati. Acad. Sci. USA Vol.85, pp. 733-737, February 1988 Biochemistry Cauliflower mosaic virus gene VI produces a symptomatic phenotype in transgenic tobacco plants (plant DNA virus/virus symptomology) GAIL A. BAUGHMAN, JERRY D. JACOBS*, AND STEPHEN H. HOWELLt Biology Department C016, University of California at San Diego, La Jolla, CA 92093 Communicated by Myron K. Brakke, October 5, 1987 (receivedfor review July 29, 1987) ABSTRACT Gene VI of the cauliflower mosaic virus inclusion body. The major inclusion body matrix protein is (CaMV) genome encodes a protein (P66) in virus-infected encoded by gene VI in the CaMV genome and is translated plants that accumulates in cytoplasmic inclusion bodies. When from the 19S RNA-one of two major RNAs produced from a segment of the CaMV genome bearing gene VI is transferred the CaMV genome. Gene VI has been implicated in host to tobacco plants by the Agrobacterium tumefaciens Ti plas- range control and in symptom production (12). We demon- mid, the resulting transgenic plants display viral-like symp- strate in this paper that transgenic tobacco plants that toms. Symptoms produced by the DNA from two different express gene VI display a symptomatic phenotype charac- viral isolates (CaMV Cabb B-JI and CM1841) were distinct- teristic of virus-infected plants. symptoms from the first were mosaic-like, whereas the other caused uniform bleaching of leaves. That gene VI was respon- MATERIALS AND METHODS sible for the symptomatic phenotype was demonstrated by Plasmid Constructions. The basic construct used in these showing that symptom production was blocked by deletions experiments was a 4840-base-pair (bp) Sal I-Xho I fragment and by a frame-shifting linker mutation in gene VI. Further- from the CaMV genome linked to a DNA fragment from the more, in primary transformants, there was a strict correlation 3' end of the nopaline synthetase (nos) gene, which provides between the appearance of symptoms and the presence of gene a poly(A) signal (13). The CaMV DNA fragment (positions VI product, P66, detected by immunoblots. Hence, a protein 4833-1642) was obtained from pLW303, derived from a encoded by the CaMV genome produces viral-like symptoms CaMV Cabb B-JI isolate (14), or pCaMV10, from a CaMV in transgenic tobacco plants. CM1841 isolate (15), and carries from left to right (Fig. 1) the distal portion of open reading frame (ORF) V (V'), the 19S The pathogenic effects of viruses on plant growth, develop- promoter, ORF VI, the 35S promoter, ORF VII, ORF I, and ment, and symptom production are described in rich detail in the proximal portion of ORF II (I'). To link the CaMV DNA the plant literature; however, the mechanisms by which to the nos 3' segment, the Sal I-Xho I fragment of the viral plant viruses cause disease are poorly understood at a genome was inserted into the Sal I site of pUC18Xnos 3', molecular level (for review, see ref. 1). Symptoms produced which carries the 1028-bp nos 3' fragment in a derivative of by plant viruses are very diverse, but a common symptom in pUC18 that has an Xho I linker inserted in the HindIII site of systemic infections is a mosaic leaf pattern that results from the polylinker. the yellowing (chlorosis, bleaching, or clearing) of vascular An Xho I-Kpn I fragment containing the linked CaMV channels that delimit "green islands" of tissue in the leaf. DNA-nos 3' segment was inserted into the Sal I-Kpn I sites What causes the yellowing and the intensity of symptoms of Bin 19, an Agrobacterium tumefaciens binary Ti-plasmid is a matter of considerable interest. In plants infected with vector (16), to create pCaMV201 (Cabb B-JI isolate) and tobacco mosaic virus (TMV) (2) and cucumber mosaic virus pGB220 (CaMV CM1841 isolate). These plasmids were (CMV) (3), virus is usually concentrated in the chlorotic transferred by conjugation to A. tumefaciens AtilO (LBA- regions, whereas the dark green islands are relatively virus- 4404 derivative) to yield At115 and At170, respectively (Fig. free. Among different isolates of the same virus, there 1). appears to be direct correlation between the severity of By making the appropriate deletions, we subdivided the symptoms (chlorosis) and the extent of TMV accumulation CaMV DNA insert into segments carrying either the 19S or (4). Attenuated TMV strains, such as the L11A strain, which the truncated 35S transcription units. Deletions were made differs from a normal strain by a single amino acid change in in pGB212, a progenitor plasmid (with pUC18 backbone) of the 183-kDa protein (5), accumulate to a lesser extent than pGB220. A Sal I linker was inserted at the EcoRV site the wild-type counterpart. However, among different vi- (CaMV map position 7342) or the Ava I site (position 6688) of ruses, the severity of symptoms does not necessarily relate the CaMV DNA insert, and the Sal I fragment containing a to the extent of virus accumulation (6). This may be due to portion of the 19S transcription unit was deleted; then the the fact that different viruses may have different means by remainder of the CaMV-nos 3' segment was inserted into Bin which they produce symptoms. 19 and transferred by conjugation into A. tumefaciens to Most isolates of cauliflower mosaic virus (CaMV) produce create At171 and At174, respectively. These two plasmids typical mosaic symptoms in the leaves of systemically in- differ by length of the region upstream from the 35S pro- fected host plants (7). At the cellular level, a prominent moter. In another construct, which gave rise to At172, cytopathological effect of CaMV infection is the appearance nearly all of the truncated 35S transcription unit was deleted of viral inclusion bodies or viroplasms in the cytoplasm of from the Asu II site (CaMV map position 7730), 295 bp infected cells. The most abundant viral-encoded protein, P66 (8-11), in infected cells is a major protein ofthe matrix ofthe Abbreviations: CaMV, cauliflower mosaic virus; TMV, tobacco mosaic virus; CMV, cucumber mosaic virus; P66, CaMV gene VI product; ORF, open reading frame. The publication costs of this article were defrayed in part by page charge *Present address: Zoology Department, University of Washington, payment. This article must therefore be hereby marked "advertisement" Seattle, WA 98195. in accordance with 18 U.S.C. §1734 solely to indicate this fact. tTo whom reprint requests should be addressed. 733 Downloaded by guest on October 2, 2021 734 Biochemistry: Baughman et al. Proc. Natl. Acad. Sci. USA 85 (1988) Asu 11 Asu 11 terminus so that it could be conjugated to carrier bovine (7730) serum albumin according to Walter et al. (21). RESULTS Transgenic tobacco plants in the Atl15, Atl70, or At172 plant series (transgenic plants transformed by Atll5, Atl70, or Atl72, respectively) bearing a 4.8-kb segment of the CaMV genome (in AtllS and At170) or a 2.2-kb subfragment (in At172, Fig. 1) displayed an interesting phenotype. The Kpn I leaves of most plants showed typical symptoms of virus (Asp 718) infection of host plants. The leaves either had a blotchy (mosaic-like) appearance (Fig. 2A) or were uniformly light green (Fig. 2B) when compared to asymptomatic plants (Fig. 2C). We have tested the same DNA segment from two different CaMV isolates and have found that the DNA from the CaMV Cabb B-JI isolate (Atll5 series) usually gave a blotchy phenotype, whereas the CaMV CM1841 isolate (At170 and At172 series) produced a more uniform light green background. The 4.8-kb CaMV DNA segment introduced into these plants is a complex region of the viral genome that was linked to a 3' fragment from the nos gene (Fig. 1). The viral DNA segment contained two transcription units-the 19S RNA unit, which encodes ORF VI, and a truncated 35S FIG. 1. Map of plasmids in Atl70 (pGB220) and Atll5 RNA unit, which included ORFs VII and I. To find out what (pCaMV201) and plasmids derived from pGB220. Plasmids in Atl7O viral determinants were responsible for the symptomatic and Atll5 were produced by inserting the 4.8-kilobase (kb) Sal phenotype, we produced various deletions in the viral DNA I-Xho I CaMV fragment (from CaMV isolates CM1841 and Cabb segment and reintroduced the constructs into plants by B-JI, respectively) linked to the 1.0-kb nos 3' fragment into the A. tumefaciens binary Ti-plasmid vector, Bin 19 (16). Atl71, At172, means of the Ti plasmid. We found that all transformed and At173 have plasmids with deletions (A) as indicated. At173 has plants were asymptomatic (Table 1) when we deleted part an 8-bp Sal I linker insert at the Bal I site at position 5940 in the (At174 series) or nearly all (Atl71 series) of the 19S RNA CaMV genome segment. RB, right border; LB, left border. transcription unit, leaving the 35S RNA transcription unit intact with about 750 bases upstream from the start of 35S downstream from the start of transcription, to the Asu II site RNA transcription in the case of the At174 series and only in the nos 3' segment. A frame-shifting linker mutation in about 90 bases upstream in the Atl71 constructs. However, CaMV gene VI was created by inserting an 8-bp Sal I linker deletion of the 35S RNA transcription unit (At172 series, at the Bal I site (CaMV map position 5940) in pGB212. This Fig. 1) had no effect on symptom production (Fig. 2B, Table construct was inserted into Bin 19 and transferred to A. 1). These results pointed to gene VI in the 19S transcription tumefaciens to create At173. unit as a possible determinant of symptom production.
Load more

Recommended publications
  • TESTS of the Associanon of HEAT SHOCK PROTEIN 90 with A
    TESTS OF THE ASSOCIAnON OF HEAT SHOCK PROTEIN 90 WITH A CAULIFLOWER MOSAIC VIRUS REVERSE TRANSCRIPTASE By BRESHANA QUIENE JOHNSON Bachelor of Science Xavier University of Louisiana New Orleans, Louisiana May 1997 Submitted to the Faculty of the Graduate College of the Oklahoma State University in partial fulfillment of the requirements for the Degree of MASTER OF SCIENCE July, 2000 Ok/ahoma State University Library TESTS OF THE ASSOCIAnON OF HEAT SHOCK PROTEIN 90 WITH A CAULIFLOWER MOSAIC VIRUS REVERSE TRANSCRIPTASE Thesis Approved: ~~-- Thesis Advisor ~~-~---- 11 ACKNOWLEDGEMENTS I sincerely thank my thesis advisor, Dr. Ulrich K. Melcher, for his guidance, supervision, expertise, and friendship. It was a pleasure to be a part of his Jab and I will always appreciate his dedication to his students and encouragement. I thank my committee members Dr. Richard C. Essenberg and Dr. Robelt L. Matts for their support and expertise. I also thank Dr. James B. Blair and the Department of Biochemistry and Molecular Biology for providing me with the opportunity to participate in the NIH Biomedical Graduate Students program and their financial support. I express my sincere gratitude to Dr. Steven D. Hartson for his guidance, suggestions, and inspiration throughout this project. I would like to thank staff and students of the Department of Biochemistry and Molecular Biology, who provided me with assistance: Ann Williams, Dr. Ba am Fraij, Dr. Jerry Merz, Wenhao Wang, Qi Jiang, Kenji Onodera, Wenjun Huang, Jieya Shao, Abdel Bior, and Thomas Prince I truly thank my parents for encouraging and supporting me throughout my academic pursuits. I am very grateful to them and blessed to have their love and concern.
    [Show full text]
  • CRISPR/Cas9-Mediated Resistance to Cauliflower Mosaic Virus Haijie Liu1
    bioRxiv preprint doi: https://doi.org/10.1101/191809; this version posted September 23, 2017. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. CRISPR/Cas9-mediated resistance to cauliflower mosaic virus Haijie Liu1,*, Cara L. Soyars2,7, *, Jianhui Li1, *, Qili Fei4,5, Guijuan He1, Brenda A. Peterson2,7, Blake C. Meyers4,5,6, Zachary L. Nimchuk2,3,7,8, and Xiaofeng Wang1,#. 1Department of Plant Pathology, Physiology and Weed Science; 2Department of Biological Sciences; 3Faculty of Health Sciences; Virginia Tech, Blacksburg, VA, USA; 4Department of Plant & Soil Sciences and Delaware Biotechnology Institute, University of Delaware, Newark, Delaware, USA; 5Donald Danforth Plant Science Center, St. Louis, Missouri, USA; 6University of Missouri – Columbia, Division of Plant Sciences, Columbia, MO; 7Department of Biology, University of North Carolina at Chapel Hill, and 8Curriculum in Genetics and Molecular Biology, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA; *These authors contributed equally to this project. # Author for correspondence: Xiaofeng Wang, 549 Latham Hall, 220 Ag Quad Lane, Blacksburg, VA 24061. Telephone: 1-540-231-1868, Fax: 1-540-231-7477, Email: [email protected]. H. Liu: [email protected]; C.L. Soyars: [email protected]; J. Li: [email protected]; Q. Fei: [email protected]; G. He: [email protected]; B.A. Peterson: [email protected]; B.C. Meyers: [email protected]; Z.L. Nimchuk: [email protected] Running title: CRISPR-Cas9-conferred resistance to CaMV Key words: CRISPR-Cas9, cauliflower mosaic virus, virus resistance, virus escape, small RNA Word count: Summary, 216 and text, 4201 1 bioRxiv preprint doi: https://doi.org/10.1101/191809; this version posted September 23, 2017.
    [Show full text]
  • Cauliflower Mosaic Virus (Camv)
    Cauliflower Mosaic Virus (CaMV) Dr.Ramesh C.K Cauliflower Mosaic Virus (CaMV) • Cauliflower mosaic virus (CaMV) is a member of the genus Caulimovirus, one of the six genera in the family Caulimoviridae which are pararetroviruses that infect plants. • Pararetroviruses group due to its mode of replication via reverse transcription of a pre- genomic RNA intermediate. just like retroviruses but the viral particles contain DNA instead of RNA. • True retroviruses are not known in plants; however, plant pararetroviruses (caulimoviridae) share many retroviral properties, replicating by transcription in the nucleus followed by reverse transcription in the cytoplasm. • Pararetroviruses have circular DNA genomes that do not integrate into the host genome, and display several unique expression strategies. • CaMV infects mostly plants of the family Brassicaceae (such as cauliflower and turnip) but some CaMV strains are also able to infect Solanaceae species of the genera Datura and Nicotiana. • CaMV induces a variety of systemic symptoms such as mosaic, necrotic lesions on leaf surfaces, stunted growth, and deformation of the overall plant structure. • CaMV is transmitted by aphid species such as Myzus persicae. Once introduced within a plant host cell, virions migrate to the nuclear envelope of the plant cell. Structure • The CaMV particle is an icosahedron with a diameter of 52 nm built from 420 capsid protein (CP), which surrounds a solvent-filled central cavity. • In addition to capsid proteins, caulimoviruses are also surrounded by virus associated proteins. These proteins are responsible for assisting in the binding of the virus to DNA • CaMV contains a circular double-stranded DNA molecule of about 8.0 kilobases, interrupted by nicks that result from the actions of RNAse H during reverse transcription.
    [Show full text]
  • Ribosome Shunting, Polycistronic Translation, and Evasion of Antiviral Defenses in Plant Pararetroviruses and Beyond Mikhail M
    Ribosome Shunting, Polycistronic Translation, and Evasion of Antiviral Defenses in Plant Pararetroviruses and Beyond Mikhail M. Pooggin, Lyuba Ryabova To cite this version: Mikhail M. Pooggin, Lyuba Ryabova. Ribosome Shunting, Polycistronic Translation, and Evasion of Antiviral Defenses in Plant Pararetroviruses and Beyond. Frontiers in Microbiology, Frontiers Media, 2018, 9, pp.644. 10.3389/fmicb.2018.00644. hal-02289592 HAL Id: hal-02289592 https://hal.archives-ouvertes.fr/hal-02289592 Submitted on 16 Sep 2019 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Distributed under a Creative Commons Attribution - ShareAlike| 4.0 International License fmicb-09-00644 April 9, 2018 Time: 16:25 # 1 REVIEW published: 10 April 2018 doi: 10.3389/fmicb.2018.00644 Ribosome Shunting, Polycistronic Translation, and Evasion of Antiviral Defenses in Plant Pararetroviruses and Beyond Mikhail M. Pooggin1* and Lyubov A. Ryabova2* 1 INRA, UMR Biologie et Génétique des Interactions Plante-Parasite, Montpellier, France, 2 Institut de Biologie Moléculaire des Plantes, Centre National de la Recherche Scientifique, UPR 2357, Université de Strasbourg, Strasbourg, France Viruses have compact genomes and usually translate more than one protein from polycistronic RNAs using leaky scanning, frameshifting, stop codon suppression or reinitiation mechanisms.
    [Show full text]
  • An Abstract of the Dissertation Of
    AN ABSTRACT OF THE DISSERTATION OF Alfredo Diaz Lara for the degree of Doctor of Philosophy in Botany and Plant Pathology presented on December 16, 2016. Title: Identification of Endogenous and Exogenous Pararetroviruses in Red Raspberry (Rubus idaeus L.) and Blueberry (Vaccinium corymbosum L.). Abstract approved: ______________________________________________________ Robert R. Martin The Pacific Northwest (Oregon and Washington in the United States and British Columbia in Canada) is one of the major producers of red raspberry (Rubus idaeus L.) and blueberry (Vaccinium corymbosum L.) in the world. The expansion of growing area with these crops has resulted in the emergence of new virus diseases that cause serious economic losses. The majority of viruses affecting plants (including blueberry and red raspberry) contain RNA genomes. In contrast, plant viruses with DNA genomes are relatively rare and most of the time ignored in virus surveys. The family Caulimoviridae is a group of plant pararetroviruses (reverse-transcribing viruses) with the ability to integrate their DNA into the host genome, resulting in complex molecular interactions that lead to inconsistencies in terms of detection and disease symptoms. Albeit, few studies have been conducted to determine the nature of plant pararetroviruses and their relationships with the associated host. To investigate the presence of pararetroviruses in blueberry and red raspberry, and their possible integration events, different plant material suspected to be infected with viruses was collected in nurseries, commercial fields and clonal germplasm repositories for a period of four years. For blueberry, using rolling circle amplification (RCA) a new virus was identified and named Blueberry fruit drop-associated virus (BFDaV) because of its association with fruit-drop disorder.
    [Show full text]
  • A New and Distinct Species in the Genus Caulimovirus Exists As an Endogenous Plant Pararetroviral Sequence in Its Host, Dahlia Variabilis
    Virology 376 (2008) 253–257 Contents lists available at ScienceDirect Virology journal homepage: www.elsevier.com/locate/yviro Rapid Communication A new and distinct species in the genus Caulimovirus exists as an endogenous plant pararetroviral sequence in its host, Dahlia variabilis Vihanga Pahalawatta, Keri Druffel, Hanu Pappu ⁎ Department of Plant Pathology, Washington State University, Pullman, WA, USA article info abstract Article history: Viruses in certain genera in family Caulimoviridae were shown to integrate their genomic sequences into their Received 6 August 2007 host genomes and exist as endogenous pararetroviral sequences (EPRV). However, members of the genus Returned to author for revision Caulimovirus remained to be the exception and are known to exist only as episomal elements in the 13 September 2007 infected cell. We present evidence that the DNA genome of a new and distinct Caulimovirus species, associated Accepted 4 March 2008 with dahlia mosaic, is integrated into its host genome, dahlia (Dahlia variabilis). Using cloned viral genes as Available online 7 May 2008 probes, Southern blot hybridization of total plant DNA from dahlia seedlings showed the presence of viral DNA in the host DNA. Fluorescent in situ hybridization using labeled DNA probes from the D10 genome localized Keywords: the viral sequences in dahlia chromosomes. The natural integration of a Caulimovirus genome into its host and Pararetrovirus – Dahlia mosaic virus its existence as an EPRV suggests the co-evolution of this plant virus pathosystem. Caulimovirus © 2008 Elsevier Inc. All rights reserved. Introduction genome, dahlia (Dahlia variabilis) and thus exists as an endogenous plant pararetrovirus (EPRV). Dahlia mosaic caulimovirus (DMV) is an important viral pathogen of dahlia in the US and several parts of the world.
    [Show full text]
  • Cauliflower Mosaic Virus P6 Dysfunctions Histone Deacetylase
    cells Article Cauliflower mosaic virus P6 Dysfunctions Histone Deacetylase HD2C to Promote Virus Infection Shun Li 1,2, Shanwu Lyu 1, Yujuan Liu 2, Ming Luo 1,3, Suhua Shi 4 and Shulin Deng 1,3,5,* 1 Guangdong Provincial Key Laboratory of Applied Botany & CAS Key Laboratory of South China Agricultural Plant Molecular Analysis and Genetic Improvement, South China Botanical Garden, Chinese Academy of Sciences, Guangzhou 510650, China; [email protected] (S.L.); [email protected] (S.L.); [email protected] (M.L.) 2 School of Life Sciences, University of Chinese Academy of Sciences, Beijing 100049, China; [email protected] 3 Center of Economic Botany, Core Botanical Gardens, Chinese Academy of Sciences, Guangzhou 510650, China 4 State Key Laboratory of Biocontrol, Guangdong Provincial Key Laboratory of Plant Resources, School of Life Sciences, Sun Yat-Sen University, Guangzhou 510275, China; [email protected] 5 National Engineering Research Center of Navel Orange, School of Life Sciences, Gannan Normal University, Ganzhou 341000, China * Correspondence: [email protected] Abstract: Histone deacetylases (HDACs) are vital epigenetic modifiers not only in regulating plant development but also in abiotic- and biotic-stress responses. Though to date, the functions of HD2C— an HD2-type HDAC—In plant development and abiotic stress have been intensively explored, its function in biotic stress remains unknown. In this study, we have identified HD2C as an interaction partner of the Cauliflower mosaic virus (CaMV) P6 protein. It functions as a positive regulator in defending against CaMV infection. The hd2c mutants show enhanced susceptibility to CaMV infection.
    [Show full text]
  • Review Article How to Deal with the Upcoming Challenges in GMO Detection in Food and Feed
    Hindawi Publishing Corporation Journal of Biomedicine and Biotechnology Volume 2012, Article ID 402418, 11 pages doi:10.1155/2012/402418 Review Article How to Deal with the Upcoming Challenges in GMO Detection in Food and Feed Sylvia R. M. Broeders, Sigrid C. J. De Keersmaecker, and Nancy H. C. Roosens Platform Biotechnology and Molecular Biology, Wetenschappelijk Instituut Volksgezondheid-Institut Scientifique de Sant´e Publique (WIV-ISP), J. Wytsmanstraat 14, 1050 Brussel, Belgium Correspondence should be addressed to Nancy H. C. Roosens, [email protected] Received 30 March 2012; Accepted 13 September 2012 Academic Editor: Joel W. Ochieng Copyright © 2012 Sylvia R. M. Broeders et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Biotech crops are the fastest adopted crop technology in the history of modern agriculture. The commercialisation of GMO is in many countries strictly regulated laying down the need for traceability and labelling. To comply with these legislations, detection methods are needed. To date, GM events have been developed by the introduction of a transgenic insert (i.e., promoter, coding sequence, terminator) into the plant genome and real-time PCR is the detection method of choice. However, new types of genetic elements will be used to construct new GMO and new crops will be transformed. Additionally, the presence of unauthorised GMO in food and feed samples might increase in the near future. To enable enforcement laboratories to continue detecting all GM events and to obtain an idea of the possible presence of unauthorised GMO in a food and feed sample, an intensive screening will become necessary.
    [Show full text]
  • Cauliflower Mosaic Virus (Camv) Biology, Management, And
    MINI REVIEW published: 31 March 2020 doi: 10.3389/fsufs.2020.00021 Cauliflower mosaic virus (CaMV) Biology, Management, and Relevance to GM Plant Detection for Sustainable Organic Agriculture Aurélie Bak and Joanne B. Emerson* Department of Plant Pathology, University of California, Davis, Davis, CA, United States In today’s global market, some organic farmers must meet regulatory requirements to demonstrate that their plants and feedstocks are genetically modified organism (GMO)-free. Many GM plants are engineered to contain a promoter from the plant virus, Cauliflower mosaic virus (CaMV), in order to facilitate expression of an engineered target gene. The relative ubiquity of this CaMV 35S promoter (P35S) in GM constructs means that assays designed to detect GM plants often target the P35S DNA sequence, but these detection assays can yield false-positives from plants that are infected by naturally-occurring CaMV or its relatives within the Caulimoviridae. This review places Edited by: CaMV infection and these ambiguous GM plant detection assays in context, serving Avtar Krishan Handa, Purdue University, United States as a resource for industry professionals, regulatory bodies, and researchers at the Reviewed by: nexus of organic farming and global commerce. We first briefly introduce GM plants James Schoelz, from a regulatory perspective, and then we describe CaMV biology, transmission, and University of Missouri, United States Tahira Fatima, management practices, highlighting the relatively widespread nature of CaMV infection Purdue University, United States in both GM and non-GM crops within the Brassicaceae and Solanaceae families. Finally, *Correspondence: we discuss current knowledge of public food safety related to the consumption of Joanne B.
    [Show full text]
  • Criteria for Identification of Cauliflower Mosaic Virus's of the Far Eastern
    Plant Protection Science – 2002 Plant Protection Science – 2002 Vol. 38, Special Issue 2: 258–260 Criteria for Identification of Cauliflower Mosaic Virus’s of the Far Eastern Strains R. V. GNUTOVA*, V. F. TOLKACH and JU. V. BOGUNOV Institute of Biology & Soil Science Far Est Branch, 690022 Vladivostok, Russia *E-mail: [email protected] Abstract On the base of the present-day principles to classify plant viruses the identification of Cauliflower mosaic virus (CaMV), a new virus for the Russian Federation, is carried out. Biological properties of 7 isolates have been studied. Symptoma- tology, range of host-plants and physical properties of virions of studied strains differ. The least thermostable strain is CaMV-B3 (TIP – 75°C) and the highest TIP (85°C) is CaMV-B1. The highest virus concentration in sap was observed for CaMV-B2 (DEP – 10–6), and lowest – CaMV-R1 (10–1–10–2). CaMV-B2 and CaMV-C2 lost infection during 4 days in room conditions, CaMV-B3 – 1 day. A virus proteins were isolated (42 and 44 kD). The native nucleic acid of CaMV have been extracted. The DNA was separated into mixtures of circular and linear molecules. Size of the DNA is about 8000 base pairs. Keywords: Cauliflower mosaic virus; Brassicaceae; identification INTRODUCTION Primorye Territory. CaMV is the first DNA-containing phytovirus, discovered in the territory of Russia. The fundamental investigations are essential for study of properties virus-specific proteins and nu- MATERIALS AND METHODS cleic acids of phytoviruses and their strains not only described in literature but also recent for description The virus isolates with virus-like symptoms were to a new species according to the latest requirement used, isolated from cauliflower (CaMV-C1, CaMV-C2), of classification.
    [Show full text]
  • The Expression of Foreign Gene Under the Control of Cauliflower Mosaic Virus 35S RNA Promoter
    Cell Research (1990), 1, 1-10 The expression of foreign gene under the control of cauliflower mosaic virus 35s RNA promoter Wang Hao and Bai Yongyan Shanghai Institute of Plant Physiology, Academia Sinica. 300 Fenglin Road, Shanghai 200032, China ABSTRACT The promoter region of cauliflower mosaic virus (CaMV) 35s RNA was employed to construct an intermediate expres- sion vector which can be used in Ti plasmid system of Agro- bacterium tumefaciens. The original plasmid, which contains a polylinker between CaMV 35s RNA and its 3' termination signal in pUC18 was modified to have another antibiotic resistance marker (kanamycin resistance gene Kmr) to facili- tate the selection of recombinant with Ti plasmid. Octopine synthase (ocs) structural gene was inserted into this vector downstream of CaMV 35s RNA promoter. This chimaeric gene was introduced into integrative Ti plasmid vector pGV3850, and then transformed into Nicotiana tobaccum cells. A binary plasmid vector was also used to introduce the chimaeric gene into tobacco cells. In both cases, the expression of ocs gene was demonstrated. The amount of oc- topine was much more than the nopaline synthesized by no- paline synthase (nos) gene transferred at the same time with Ti plasmid vector. This demonstrated that CaMV 35s RNA promoter is stronger in transcriptional function than the pro- moter of nos in tobacco cells. Key words: Agrobacterium tumefaciens, gene expression, cauliflower mosaic Cirus 35s RNA promoter. INTRODUCTION During the past decade, extensive research on the Ti plasmid of Agrobacterium tumefaciens has brought development of useful and efficient vectors to introduce fo- reign genes into dicotyledonous plants.
    [Show full text]
  • Replication of Cauliflower Mosaic Virus ORF I Mutants in Turnip Protoplasts
    日木直病 報 60: 27-35 (1994) Ann. Phytopath. Soc. Japan 60: 27-35 (1994) Replication of Cauliflower Mosaic Virus ORF I Mutants in Turnip Protoplasts Seiji TSUGE*,•õ, Kappei KOBAYASHI*,•õ•õ, Hitoshi NAKAYASHIKI*, Tetsuro OKUNO* and Iwao FURUSAWA* Abstract We succeeded in infecting turnip protoplasts with a cloned cauliflower mosaic virus (CaMV) DNA, pCa122, which contains 1.2 copy of CaMV genomic DNA and a plasmid expressing open reading frame (ORF) VI products (pEXP6) using polyethylene glycol. Fluorescent antibody staining showed that up to 50% of protoplasts were infected. It was difficult to detect the progeny DNA and the viral protein in protoplasts inoculated with pCa122 alone. Co-transfection with the plasmid pEXP6 produced larger fluorescing specks in each infected protoplasts and increased the accumulation of the progeny DNA and some other viral proteins to detectable levels. Using this protoplast system, three CaMV ORF I insertional mutants which were not infectious on turnip plants were tested for their infectivity on turnip protoplasts. Viral DNA and products accumulated in infected proto- plasts to the same extent of the wild type DNA-infected protoplasts. These results indicate that ORF I product is not required for multiplication of CaMV in protoplasts, but is indispensable for infection on whole plants, strongly supporting that ORF I product is involved in cell-to-cell movement of CaMV. (Received May 6, 1993) Key words: cauliflower mosaic virus, protoplasts, movement protein. INTRODUCTION Cauliflower mosaic virus (CaMV) has an 8kb circular double stranded DNA genome which encodes six major open reading frames (ORFs) I to VI on the same DNA strand17).
    [Show full text]