Skip to main content
Log in

Cauliflower mosaic virus gene VI causes growth suppression, development of necrotic spots and expression of defence-related genes in transgenic tobacco plants

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

In order to study possible functions of the inclusion body matrix protein (IBMP) encoded by gene VI of cauliflower mosaic virus (CaMV), the XbaI fragment containing the gene VI of a Japanese strain of CaMV (CaMV S-Japan) was transferred to tobacco plants by Ti mediated transformation. Eight out of 18 kanamycin resistant plants (40%) expressed detectable levels of IBMP. Those transgenic plants expressing IBMP produced leaves with light green color, and their growth was suppressed as compared with control plants. Symptom-like necrotic spots also appeared on the leaves and stems of the mature transgenic plants. Furthermore, in these transgenic plants, pathogenesis-related proteins 1a, 1b and 1c were highly expressed and the activity of 1,3-β-glucanase was increased up to eightfold. From these results, we concluded that expression of the IBMP is associated with symptom development.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • An G (1986) Development of plant promoter expression vectors and their use for analysis of differential activity of nopaline synthase promoter in transformed tobacco cells. Plant Physiol 81:86–91

    Google Scholar 

  • Balazs E, Guilley H, Jonard G, Richards K (1982) Nucleotide sequence of DNA from an altered-virulence isolate D/H of the cauliflower mosaic virus. Gene 19:239–249

    Google Scholar 

  • Baughman GA, Jacobs JD, Howell SH (1988) Cauliflower mosaic virus gene VI produces a symptomatic phenotype in transgenic tobacco plants. Proc Natl Acad Sci USA 85:733–737

    Google Scholar 

  • Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Google Scholar 

  • Covey SN, Hull R (1981) Transcription of cauliflower mosaic virus DNA. Detection of transcripts, properties, and location of the gene encoding the virus inclusion body protein. Virology 111:463–474

    Google Scholar 

  • Daubert S, Richins R, Shepherd RJ, Gardner RC (1982) Mapping of the coat protein gene of cauliflower mosaic virus by its expression in a prokaryotic system. Virology 122:444–449

    Google Scholar 

  • Daubert S, Shepherd RJ, Gardner RC (1983) Insertional mutagenesis of the cauliflower mosaic virus genome. Gene 25:201–208

    Google Scholar 

  • Daubert S, Schoelz JE, Debao L, Shephard RJ (1984) Expression of disease symptoms in cauliflower mosaic virus genomic hybrids. J Mol Appl Genet 2:537–547

    Google Scholar 

  • Dixon LK, Hohn T (1984) Initiation of translation of the cauliflower mosaic virus genome from a polycistronic mRNA: evidence from deletion mutagenesis. EMBO J 3:2731–2736

    Google Scholar 

  • Dixon LK, Koenig I, Hohn T (1983) Mutagenesis of cauliflower mosaic virus. Gene 25:189–199

    Google Scholar 

  • Franck A, Guilley H, Jonard G, Richards K, Hirth L (1980) Nucleotide sequence of cauliflower mosaic virus DNA. Cell 21:285–294

    Google Scholar 

  • Gardner RC, Howarth AJ, Hahn P, Brown-Luedi M, Shepherd RJ, Messing J (1981) The complete nucleotide sequence of an infectious clone of cauliflower mosaic virus by M13mp7 shotgun sequencing. Nucleic Acids Res 9:2871–2888

    Google Scholar 

  • Giband M, Mesnard JM, Lebeurier G (1986) The gene III product (P15) of cauliflower mosaic virus is a DNA-binding protein while an immunologically related P11 polypeptide is associated with virions. EMBO J 5:2433–2438

    Google Scholar 

  • Givord L, Xiong C, Giband M, Koenig I, Hohn T, Lebeurier G, Hirth L (1984) A second cauliflower mosaic virus gene product influences the structure of the viral inclusion body. EMBO J 3:1423–1427

    Google Scholar 

  • Goldberg KB, Young MJ, Schoelz JE, Kiernan JM, Shepherd RJ (1988) Single gene of CaMV induces disease. Phytopathology 77:1704

    Google Scholar 

  • Gordon K, Pfeiffer P, Futterer J, Hohn T (1988) In vitro expression of cauliflower mosaic virus genes. EMBO J 7:309–317

    Google Scholar 

  • Hills GJ, Campbell RN (1968) Morphology of broccoli necrotic yellows virus. J Ultrastruct Res 24:134–144

    Google Scholar 

  • Hirochika H, Takatsuji H, Ubasawa A, Ikeda, J-E (1985) Site-specific deletion in cauliflower mosaic virus DNA: possible involvement of RNA splicing and reverse transcription. EMBO J 4:1673–1680

    Google Scholar 

  • Hirth L (1986) The molecular biology of caulimoviruses. Microbiol Sci 3:260–265

    Google Scholar 

  • Hoekema A, Hirsch PR, Hooykaas PJJ, Schilperoort RA (1983) A binary plant vector strategy based on separation of vir-and T-region of the Agrobacterium tumefaciens Ti-plasmid. Nature 303:179–180

    Google Scholar 

  • Hohn T, Richards K, Lebeurier G (1982) Cauliflower mosaic virus on its way to becoming a useful plant vector. Curr Top Microbiol Immunol 96:194–236

    Google Scholar 

  • Hohn T, Hohn B, Pfeiffer P (1985) Reverse transcription in CaMV. Trends Biochem Sci 10:205–209

    Google Scholar 

  • Howell SH (1982) Plant molecular vehicles: Potential vectors for introducing foreign DNA into plants. Annu Rev Plant Physiol 33:609–650

    Google Scholar 

  • Kauffmann S, Legrand M, Geoffroy P, Fritig B (1987) Biological function of ‘pathogenesis-related’ proteins: four PR proteins of tobacco have 1,3-β-glucanase activity. EMBO J 6:3209–3212

    Google Scholar 

  • Koenig R (1981) Indirect ELISA methods for the broad specificity detection of plant viruses. J Gen Virol 55:53–62

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacterial phage T4. Nature 227: 680–685

    Google Scholar 

  • Legrand M, Kauffmann S, Geoffroy P, Fritig B (1987) Biological function of pathogenesis-related proteins: Four tobacco pathogenesis-related proteins are chitinases. Proc Natl Acad Sci USA 84:6750–6754

    Google Scholar 

  • Martinez-Izquierdo JA, Hohn T (1987) Cauliflower mosaic virus coat protein is phosphorylated in vitro by a virion-associated protein kinase. Proc Natl Acad Sci USA 84:1824–1828

    Google Scholar 

  • Martinez-Izquierdo JA, Futterer J, Hohn T (1987) Protein encoded by ORF I of cauliflower mosaic virus is part of the viral inclusion body. Virology 160:527–530

    Google Scholar 

  • Mazzolini L, Bonneville JM, Volovitch M, Magazin M, Yot P (1985) Strand-specific viral DNA synthesis in purified viroplasms isolated from turnip leaves infected with cauliflower mosaic virus. Virology 145:293–303

    Google Scholar 

  • Menissier J, Laquel P, Lebeurier G, Hirth L (1984) A DNA polymerase activity is associated with cauliflower mosaic virus. Nucleic Acids Res 12:8769–8778

    Google Scholar 

  • Modjtahedi N, Volovitch M, Sossountzov L, Habricot Y, Bonneville JM, Yot P (1984) Cauliflower mosaic virus-induced viroplasms support viral DNA synthesis in a cell-free system. Virology 133:289–300

    Google Scholar 

  • Morrissey JH (1981) Silver stain for proteins in polyacrylamide gels: A modified procedure with enhanced uniform sensitivity. Anal Biochem 117:307–310

    Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassay with tobacco tissue cultures. Physiol Plant 15:473–497

    Google Scholar 

  • Odell JT, Howell SH (1980) The identification, mapping, and characterization of mRNA for P66, a cauliflower mosaic viruscoded protein. Virology 102:349–359

    Google Scholar 

  • Ohashi Y, Matsuoka M (1985) Synthesis of stress proteins in tobacco leaves. Plant Cell Physiol 26:473–480

    Google Scholar 

  • Pfeiffer M, Laquel P, Hohn T (1984) Cauliflower mosaic virus replication complexes: characterization of the associated enzymes and of the polarity of the DNA synthesized in vitro. Plant Mol Biol 3:261–270

    Google Scholar 

  • Schoelz JE, Shepherd RJ (1988) Host range control of cauliflower mosaic virus. Virology 162:30–37

    Google Scholar 

  • Schoelz JE, Shepherd RJ, Daubert S (1986) Region VI of cauliflower mosaic virus encodes a host range determinant. Mol Cell Biol 6:2632–2637

    Google Scholar 

  • Shepherd RJ (1979) DNA plant viruses. Annu Rev Plant Physiol 30:405–423

    Google Scholar 

  • Shewmaker CK, Caton JR, Houck CM, Gardner RC (1985) Transcription of cauliflower mosaic virus integrated into plant genomes. Virology 140:281–288

    Google Scholar 

  • Takatsuji H, Hirochika H, Fukushi T, Ikeda J-E (1986) Expression of cauliflower mosaic virus reverse transcriptase in yeast. Nature 319:240–243

    Google Scholar 

  • Toh H, Hayashida H, Miyata T (1983) Sequence homology between retroviral reverse transcriptase and putative polymerases of hepatitis B virus and cauliflower mosaic virus. Nature 305:827–829

    Google Scholar 

  • Towbin H, Staehlin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354

    Google Scholar 

  • van Loon LC (1985) Pathogenesis-related proteins. Plant Mol Biol 4:111–116

    Google Scholar 

  • Volovitch M, Modjtahedi N, Yot P, Brun G (1984) RNA-dependent DNA polymerase activity in cauliflower mosaic virus-infected plant leaves. EMBO J 3:309–314

    Google Scholar 

  • Woolston CJ, Covey SN, Penswick JR, Davies JW (1983) Aphid transmission and a polypeptide are specified by a defined region of the cauliflower mosaic virus genome. Gene 23:15–23

    Google Scholar 

  • Xiong C, Muller S, Lebeurier G, Hirth L (1982) Identification by immuno-precipitation of cauliflower mosaic virus in vitro major translation product with a specific serum against viroplasm protein. EMBO J 1:971–976

    Google Scholar 

  • Young M, Shewmaker CK, Daubert S, Shepherd RJ (1987) Expression of cauliflower mosaic virus proteins from an integrated form of the viral genome. J Gen Virol 68:3217–3222

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by J. Schell

Rights and permissions

Reprints and permissions

About this article

Cite this article

Takahashi, H., Shimamoto, K. & Ehara, Y. Cauliflower mosaic virus gene VI causes growth suppression, development of necrotic spots and expression of defence-related genes in transgenic tobacco plants. Mol Gen Genet 216, 188–194 (1989). https://doi.org/10.1007/BF00334355

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00334355

Key words

Navigation