Abstract
A bacterial rnc gene coding for a double-stranded RNA-dependent RNase III endoribonuclease and a mutant, rnc70, were expressed in tobacco plants. The RNase III protein produced in the transgenic plants was the same size as the bacterial protein. Expression of the wild-type gene could cause stunting in some plant lines, but not in others. Expression of the mutant protein did not affect normal growth and development of the transgenic plants. Transgenic plants of the R1 and R2 generations, expressing the wild type, as well as a mutant protein, were resistant to infection by three disparate RNA plant viruses with a divided genome but not against two viruses with a single-stranded RNA genome. Introduction of the rnc gene in crop plants may provide resistance to economically important virus diseases.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Alluvia S, Locker-Gelada H, Kobe S, Ben-Num O, Oppenheim AB: Rnase III stimulates the translation of the eIII gene of bacteriophage lambda. Proc Natl Acad Sci USA: 83: 5175–5179 (1987).
An G, Ebert PR, Mitra A, Ha SB: Binary Vectors. In: Gelvin SB, Verma DPS, Schilperoort RA Plant Molecular Biology Manual, pp. A3: 1–19. Kluwer Academic Publishers, Dordrecht, Netherlands (1988).
Bradford M: 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 (1976).
Casadaban MJ, Cohen SN: Analysis of gene control signals by DNA fusion and cloning in Escherichia coli. J Mol Biol 138: 179–207 (1980).
Chen SM, Takiff HE, Barber AM, Dubois GC, Bardwell JCA, Court D: Expression and characterization of RNase III and Era proteins: products of the rnc operon of Escherichia coli. J Biochem 265: 2888–2895 (1990).
Cheo PC: Subliminal infection of cotton by tobacco mosaic virus. Phytopathology 60: 41–46 (1970).
Court D: RNA processing and degradation by RNase III in control of messenger RNA stability, pp. 71–116. Brawerman G, Belasco J Academic Press, New York (1993).
Falk BW, Bruening G: Will transgenic crops generate new viruses and new diseases? Science 263: 1395–1396 (1994).
Feinberg AP, Vogelstein B: A technique for radio labeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132: 6–10 (1983).
Finnegan PM, Brown CG: Autonomously replicating RNA in mitochondria ofmaize plants with S-type cytoplasm. Proc Natl Acad Sci USA 83: 5175–5179 (1986).
Gerdes K, Nielsen A, Thorsted P, Wagner EGH: Mechanism of killer gene activation: antisense RNA-dependent RNase III cleavage ensures rapid turnover of the stable Hok, ShrB and PndA effector messenger RNAs. J Mol Biol 226: 637–649 (1992).
Greene AE, Allison RF: Recombination between viral RNA and transgenic plant transcripts. Science 263: 1423–1425 (1994).
Guillemaut P, Maréchal-Drouard L: Isolation of plant DNA: A fast, inexpensive, and reliable method. Plant Mol Biol Rep 10: 60–65 (1992).
Hoekema A, Hirsch PR, Hooykaas PJJ, Schilperoort RA: A binary plant vector strategy based on separation of vir-and Tregion of the Agrobacterium tumefaciens Ti-plasmid. Nature 303: 179–181 (1983).
Horsch RB, Fry JE, Hoffman NL, Eichholtz D, Rogers SG, Fraley RT: A simple and general method for transferring genes into plants. Science 227: 1229–1231 (1985).
Hull R, Davies JW: Approaches to nonconventional control of plant virus disease. Crit Rev Plant Sci 11: 17–33 (1992).
Kameyama L, Fernandez L, Court D, Guarneros G: RNase III activation of bacteriophage lambda N synthesis. Mol Microbiol 5: 2953–2963 (1991).
Köohn B, Guerra DG, Mowry TM, Berger PH: A yeast-derived RNase that confers resistance to multiple plant viruses. Phytopathology 85: 1212 (1995).
Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage. Nature 227: 680–685 (1970).
Libonati M, Carsana A, Furia A: Double stranded RNA. Mol Cell Biochem 31: 147–164 (1980).
Logemann J, Schell J, Willmitzer L: Improved method for the isolation of RNAform plant tissues. Anal Biochem 163: 16–20 (1987).
March PE, Gonzalez MA: Characterization of the biochemical properties of recombinant ribonuclease III. Nucl Acids Res 18: 3293–3298 (1990).
Matthews REF: Plant Virology, 3rd ed. Academic Press, San Diego, CA (1991).
Mitra A, An G: Three distinct regulatory elements comprise the upstream promoter region of the nopaline synthase gene. Mol Gen Genet 215: 294–299 (1989).
Mitra A, Choi HK, An G: Structural and functional analysis of Arabidopsis thaliana chlorophyll a/b-binding protein (cab) promoters. Plant Mol Biol 12: 169–179 (1989).
Murashige T, Skoog F: A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15: 473–497 (1962).
Nameth ST, JA Dodds: Double-stranded RNA detected in cucurbit varieties not inoculated with viruses. Phytopathology 75: 1293–1298 (1985).
Pines O, Yoon H-T, Inouye M: Retroregulation of the bacteriophage lambda int gene: Limited secondary degradation of the RNase III-processed transcript. J Bact 171: 588–592 (1988).
Presting GG, Smith OP, Brown CR: Resistance to potato leaf roll virus in potato plants transformed with the coat protein gene or with vector control constructs. Phytopathology 85: 436–442 (1995).
Sanford JC, Johnston SA: The concept of parasite derived resistance deriving from the parasite's own genome. J Theor Biol 113: 395–405 (1985).
Smith HA, Powers H, Swaney S, Brown C, Dougherty WG: Transgenic potato virus Y resistance in potato: evidence for an RNA-mediated cellular response. Phytopathology 85: 864–870 (1995).
Southern EM: Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98: 503–517 (1975).
Sulziniski MA, M. Zaitlin: Tobacco mosaic virus replication in resistant and susceptible plants: In some resistant species virus is confined to a small of initially infected cells. Virology 121: 12–19 (1983).
Takiff HE, Chen S-M, Court D: Genetic analysis of the rnc operon of Escherichia coli. J Bact 171: 2581–2590 (1989).
Tepfer M: Viral genes and transgenic plants. What are the potential environmental risks? Bio/technology 11: 1125–1130 (1993).
Valverde RA, Namath S, Abdallha O, Al-Musa O, Desjardins P, Dodds A: Indigenous double-stranded RNA from pepper (7Capsicum annuum). Plant Sci 61: 227–234 (1990).
Wakarchuk DA, Hamilton RI: Cellular double-stranded RNA in Phaseolous vulgaris. Plant Mol Biol 5: 55–63 (1985).
Wakarchuk DA, Hamilton RI: Partial nucleotide sequences from enigmatic ds RCAS in Phaseolous vulgaris. Plant Mol Biol 14: 637–639 (1990).
Wang Z-Y, Zheng F-Q, Guo X-L, Hong MM: Occurrence and partial characterization of an RNA species in rice mitochondria. Plant Sci 61: 227–234 (1989).
Watanabe Y, Ogawa T, Takahashi H, Ishida I, Takenchi Y, Yamamoto M, Okada Y: Resistance against multiple plant viruses in plants mediated by a double stranded-RNA specific ribonuclease. FEBS Lett 372: 165–168 (1995).
White KA, Morris TJ: Recombination between defective tombusvirus RCAS generates functional hybrid genomes. Proc Natl Acad Sci USA 91: 3642–3646 (1994).
Wilson TMA: Strategies to protect crop plants against viruses: pathogen-derived resistance blossoms. Proc Natl Acad Sci USA 90: 3134–3141 (1993).
Yanisch-Perron C, Vieira J, Messing J: Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13 mp18 and puc19 vectors. Gene 33: 113–119 (1985).
Zabalgogeazcoa IA, Gildow EF: Double-stranded ribonucleic acid in ‘Barsoy’ barley. Plant Sci 83: 187–194 (1992).
Zaccomer B, Haenni A-L, Macaya G: The remarkable variety of plant RNA virus genomes. J Gen Virol 76: 231–247 (1995).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Langenberg, W.G., Zhang, L., Court, D.L. et al. Transgenic tobacco plants expressing the bacterial mc gene resist virus infection. Molecular Breeding 3, 391–399 (1997). https://doi.org/10.1023/A:1009697507261
Issue Date:
DOI: https://doi.org/10.1023/A:1009697507261