The Manipulation of Plant Gene Expression Using Antisense RNA

  • Wolfgang Schuch
Part of the Plant Gene Research book series (GENE)


Gene cloning techniques have allowed us to analyse in considerable detail the structure of plant genes. The availability of probes for plant genes has opened the way for detailed studies on the temporal and spatial expression of plant genes during the development of different plant organs and tissues (Kuhlemeier et al., 1987). Functional analysis of isolated plant genes was made possible through the development of gene transfer techniques (Weising et al., 1988). This has led to the detailed description of expression patterns exhibited by gene promoter sequences, and to the delineation of DNA sequences which are required for the temporal and spatial control of gene expression.


Chloramphenicol Acetyl Transferase Antisense Gene Plant Gene Expression Chloramphenicol Acetyl Transferase Activity Phosphinothricin Acetyl Transferase 
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  1. Beachy RN, Stark DM, Deom CM, Oliver MJ, Fraley RT (1987) Expression of sequences of tobacco mosaic virus in transgenic plants and their role in disease resistance. Basic Life Sci 41: 169–180Google Scholar
  2. Bird CR, Smith CJS, Ray JA, Moureau P, Bevan MW, Bird AS, Hughes S, Morris PC, Grierson D, Schuch W (1988) The tomato polygalacturonase gene and ripening-specific expression in transgenic tomato plants. Plant Mol Biol 11: 651–662CrossRefGoogle Scholar
  3. Brady C, MacAlpine G, McGlasson WB, Ueda Y (1982) Polygalacturonase in tomato fruit and the induction of ripening. Aust J Plant Physiol 9: 171–178CrossRefGoogle Scholar
  4. Cornelissen M, Vandewiele M (1989) Both RNA level and translation efficiency are reduced by antisense RNA in transgenic tobacco. Nucleic Acids Res 17: 833–843PubMedCrossRefGoogle Scholar
  5. Crookes PR, Grierson D (1983) Ultrastructure of tomato fruit ripening and the role of polygalacturonase isoenzymes in cell wall degradation. Plant Physiol 72: 1088–1093PubMedCrossRefGoogle Scholar
  6. Delauney AJ, Tabaeizadeh Z, Verma DPS (1988) A stable bifunctional antisense transcript inhibiting gene expression in transgenic plants. Proc Natl Acad Sci USA 85: 4300–4304PubMedCrossRefGoogle Scholar
  7. DellaPenna D, Alexander DC, Bennet AB (1986) Molecular cloning of tomato fruit polygalacturonase: analysis of polygalacturonase mRNA levels during ripening. Proc Natl Acad Sci USA 83: 6420–6424PubMedCrossRefGoogle Scholar
  8. Ecker JR, Davis RW (1986) Inhibition of gene expression in plant cells by expression of antisense RNA. Proc Natl Acad Sci USA 83: 5372–5376PubMedCrossRefGoogle Scholar
  9. Finkelstein R, Estelle M, Martinez-Zapater J, Somerville C (1988) Arabidopsis as a tool for the identification of genes involved in plant development. In: Verma DPS, Goldberg RB (eds) Temporal and spatial regulation of plant genes. Springer, Wien New York, pp 1–25 [Dennis ES et al (eds) Plant gene research. Basic knowledge and application]Google Scholar
  10. Green PJ, Pines O, Inouye M (1986) The role of antisense RNA in gene regulation. Annu Rev Biochem 55: 569–597PubMedCrossRefGoogle Scholar
  11. Grierson D, Tucker GA (1983) Timing of ethylene and polygalacturonase synthesis in relation to the control of tomato ripening. Planta 157: 174–179CrossRefGoogle Scholar
  12. Grierson D, Tucker GA, Keen J, Bird CR, Schuch W (1986) Sequencing and identification of a cDNA clone for tomato polygalacturonase. Nucleic Acids Res 14: 8595–8603PubMedCrossRefGoogle Scholar
  13. Harland R, Weintraub H (1985) Translation of mRNA injected into Xenopus oocytes is specifically inhibited by antisense RNA. J Cell Biol 101: 1094–1099PubMedCrossRefGoogle Scholar
  14. Hastie ND, Held WA (1978) Analysis of mRNA populations by cDNA—mRNA hybrid-mediated inhibition of cell-free protein synthesis. Proc Natl Acad Sci USA 75: 1217–1221PubMedCrossRefGoogle Scholar
  15. Hemenway C, Fang R-X, Kaniewski WK, Chua N-H, Tumer NE (1988) Analysis of the mechanism of protection in transgenic plants-expressing the potato virus X coat protein or its antisense RNA. EMBO J 7: 1273–1280PubMedGoogle Scholar
  16. Herskowitz I (1987) Functional inactivation of genes by dominant negative mutations. Nature 329: 219–222PubMedCrossRefGoogle Scholar
  17. Hobson GE (1964) Polygalacturonase in normal and abnormal tomato fruit. Biochem J 92: 324–332PubMedGoogle Scholar
  18. Hobson GE (1965) The firmness of tomato fruit in relation to polygalacturonase activity. J Horticult Sci 40: 66–72Google Scholar
  19. Holdsworth MJ, Bird CR, Ray J, Schuch W, Grierson D (1987) Structure and expression of an ethylene related mRNA from tomato. Nucleic Acids Res 15: 731–739PubMedCrossRefGoogle Scholar
  20. Holdsworth MJ, Schuch W, Grierson D (1988) Organisation and expression of a wound/ripening related small multigene family from tomato. Plant Mol Biol 11: 81–88CrossRefGoogle Scholar
  21. Izant JG, Weintraub H (1984) Inhibition of thymidine kinase gene expression by antisense RNA: a molecular approach to genetic analyses. Cell 36: 1007–1015PubMedCrossRefGoogle Scholar
  22. Loesch-Fries LS, Halk E, Merlo D, Jarvis N, Nelson S, Krahn K, Burhop L, Arntzen CJ, Ryan C (1987) Molecular strategies for crop protection, expression of alfalfa mosaic virus coat protein gene and antisense cDNA in transformed tobacco tissue. UCLA Symp Proc Mol Cell Biol NS 48: 221–234Google Scholar
  23. Kuhlemeier C, Green PJ, Chua NH (1987) Regulation of gene expression in higher plants. Annu Rev Plant Physiol 38: 221–57CrossRefGoogle Scholar
  24. Paterson BM, Roberts BE, Kuff EL (1977) Structural gene identification and mapping by DNA—mRNA hybrid-arrest cell-free translation. Proc Natl Acad Sci USA 74: 4370–4374PubMedCrossRefGoogle Scholar
  25. Pines O, Inouye M (1986) Antisense RNA regulation in prokaryotes. Trends Genet 2: 284–287CrossRefGoogle Scholar
  26. Ray J, Knapp JE, Grierson D, Bird C, Schuch W (1988) Identification and sequence determination of a cDNA clone for tomato pectin esterase. Eur J Biochem 174: 119–124PubMedCrossRefGoogle Scholar
  27. Rezaian AM, Skene KGM, Ellis JG (1988) Antisense RNAs of cucumber mosaic virus in transgenic plants assessed for control of the virus. Plant Mol Biol 11: 463–471CrossRefGoogle Scholar
  28. Robert LS, Donaldson PA, Ladaique C, Altosaar I, Arnison PG, Fabijanski SF (1989) Antisense RNA inhibition of ß-glucuronidase gene expression in transgenic tobacco plants. Plant Mol Biol 13: 399–409PubMedCrossRefGoogle Scholar
  29. Rodermel SR, Abbott MS, Bogorad L (1988) Nuclear—organelle interactions: nuclear antisense gene inhibits ribulose bisphosphate carboxylase enzyme levels in transformed tobacco plants. Cell 55: 673–681PubMedCrossRefGoogle Scholar
  30. Rothstein SJ, DiMaio J, Strand M, Rice D (1987) Stable and heritable inhibition of the expression of nopaline synthase in tobacco expressing antisense RNA. Proc Natl Acad Sci USA 84: 8439–8443PubMedCrossRefGoogle Scholar
  31. Sandler SJ, Stayton M, Townsend JA, Ralston ML, Bedbrook JR, Dunsmuir P (1988) Inhibition of gene expression in transformed plants by antisense RNA. Plant Mol Biol 11: 301–310CrossRefGoogle Scholar
  32. Schuch W, Bird CR, Ray J, Smith CJS, Watson CF, Morris PC, Gray JE, Arnold C, Seymour GB, Tucker GA, Grierson D (1989) Control and manipulation of gene expression during tomato fruit ripening. Plant Mol Biol 13: 303–311PubMedCrossRefGoogle Scholar
  33. Seymour GB, Harding SE (1987) Molecular size of tomato (Lycopersicon esculentum Mill) fruit polyuronides by gel filtration and low-speed sedimentation equilibrium. Biochem J 245: 463–466PubMedGoogle Scholar
  34. Sheehy RE, Pearson J, Brady CJ, Hiatt WR (1987) Molecular characterisation of tomato fruit polygalacturonase. Mol Gen Genet 208: 30–36CrossRefGoogle Scholar
  35. Sheehy RE, Kramer MK, Hiatt WR (1988) Reduction of polygalacturonase activity in tomato fruit by antisense RNA. Proc Natl Acad Sci USA 85: 8805–8809PubMedCrossRefGoogle Scholar
  36. Simons RW, Kleckner N (1988) Biological regulation by antisense RNA in prokaryotes. Annu Rev Genet 22: 567–600PubMedCrossRefGoogle Scholar
  37. Slater A, Maunders MJ, Edwards K, Schuch W, Grierson D (1985) Isolation and characterisation of cDNA clones for tomato proteins. Plant Mol Biol 5: 137–147CrossRefGoogle Scholar
  38. Smith CJS, Slater A, Grierson D (1986) Rapid appearance of an mRNA correlated with ethylene synthesis encoding a protein of molecular weight 35,000. Planta 168: 94–100CrossRefGoogle Scholar
  39. Smith CJS, Watson CF, Ray J, Bird CR, Morris PC, Schuch W, Grierson D (1988) Antisense RNA inhibition of polygalacturonase gene expression in tomatoes. Nature 334: 724–726CrossRefGoogle Scholar
  40. Smith CJS, Watson CF, Morris PC, Bird CR, Seymour GB, Gray JE, Arnold C, Tucker GA, Schuch W, Grierson D (1990) Inheritance and effect on ripening of antisense polygalacturonase genes in transgenic tomatoes. Plant Mol BiolGoogle Scholar
  41. Smithies O, Gregg RG, Boggs SS, Koralewski MA, Kucherlapati RS (1985) Insertion of DNA sequences into the human chromosomal /i-globin locus by homologous recombination. Nature 317: 230–234PubMedCrossRefGoogle Scholar
  42. Thomas H, Grierson D (1987) Developmental mutants in higher plants. Cambridge University Press, CambridgeGoogle Scholar
  43. Tucker GA, Grierson D (1982) Synthesis of polygalacturonase during tomato fruit ripening. Planta 155: 64–67CrossRefGoogle Scholar
  44. van der Krol AR, Lenting PE, Veenstra J, van der Meer IM, Koes RE, Gerats AGM, Mol JNM, Stuitje AR (1988a) An antisense chalcone synthase gene in transgenic plants inhibits flower pigmentation. Nature 333: 866–869CrossRefGoogle Scholar
  45. van der Krol AR, Mol JNM, Stuitje AR (1988b) Modulation of eukaryotic gene expression by complementary RNA or DNA sequences. BioTechniques 6: 958–976PubMedGoogle Scholar
  46. Weising K, Schell J, Kahl G (1988) Foreign genes in plants: transfer, structure, expression, and applications. Annu Rev Genet 22: 421–477PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag/Wien 1991

Authors and Affiliations

  • Wolfgang Schuch
    • 1
  1. 1.ICI Seeds, Plant Biotechnology SectionJealott’s Hill Research StationBracknell, Berks.UK

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