The Molecular Biology of Fruit Ripening

  • Steve Picton
  • Julie E. Gray
  • Don Grierson
Part of the NATO ASI Series book series (volume 81)


The past 15 years has witnessed an explosion in the number of research papers utilising techniques of molecular biology to answer questions about the developmental regulation of gene expression in higher plants. This paper will draw on research in the authors’ laboratory utilizing the tomato (Lycopersicon esculentum) as a model plant and the process of fruit ripening as a clearly defined developmental system which has been used to examine the regulation and biochemical function of specific changes in gene expression [for review see 9]. We shall provide an overview of the development of techniques for isolation, identification and subsequent manipulation of expression of individual genes implicated in the process of fruit ripening and show how manipulation of individual genes is of use both in fundamental and applied research on food crops.


Transgenic Plant Tomato Fruit Ethylene Biosynthesis Transgenic Tomato Antisense Gene 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Biale JB, Young RE: Respiration and ripening in fruits-retrospect and prospect. In: Friend J, Rhodes MJC (eds) Recent Advances in the Biochemistry of Fruit and Vegetables, pp 1–39 Academic Press, London (1981).Google Scholar
  2. 2.
    Biggs MS, Harriman RW, Handa AK: Changes in gene expression tomato fruit ripening. Plant Physiol. 81: 395–403 (1986).PubMedCrossRefGoogle Scholar
  3. 3.
    Bird CR, Smith CJS, Moreau P, Bevan MW, Bird AS, Hughes S, Morris PC, Grierson D, Schuch W: The tomato polygalacturonase gene and ripening specific expression in transgenic plants. Plant Mol. Biol. 11: 651–662 (1988).CrossRefGoogle Scholar
  4. 4.
    Bird CR, Ray JA, Fletcher JD, Boniwell JM, Bird AS, Teulieres C, Blain I, Bramley PM, Schuch W: Using antisense RNA to study gene function: Inhibition of carotenoid biosynthesis in transgenic tomatoes. Bio-technology 9: 635–639 (1991).Google Scholar
  5. 5.
    Bouzayen M, Hamilton AJ, Picton S, Barton S, Grierson D: Identification of genes for the ethylene-forming enzyme and inhibition of ethylene synthesis in transgenic plants using antisense genes. Biochem.Soc.Trans. 20: 76–79 (1992).PubMedGoogle Scholar
  6. 6.
    Crookes PR, Grierson D: Ultrastructure of tomato fruit ripering and the role of polygalacturonase isoenzymes in cell wall degradation. Plant Physiol. 72: 1088–1093 (1983).PubMedCrossRefGoogle Scholar
  7. 7.
    Fray R, Grierson D: Identification and genetic analysis of normal and mutant phytoene synthase genes of tomato by sequencing, complementation and co-supression. Plant Mol. Biol. In Press (1993).Google Scholar
  8. 8.
    Giovanoni JJ, DellaPenna D, Bennet AB, Fischer RL: Expression of a chimeric polygalacturonase gene in transgenic rin (ripening inhibitor) tomato fruit results in polyuronide degradation but not fruit softening. Plant Cell. 1: 53–63 (1989).CrossRefGoogle Scholar
  9. 9.
    Gray J, Picton S, Shabeer J, Grierson D: Molecular biology of fruit ripening and its manipulation with antisense genes. Plant Mol. Biol. 19: 69–87 (1992).PubMedCrossRefGoogle Scholar
  10. 10.
    Grierson D, Maunders MJ, Slater A, Ray J, Bird CR, Schuch W, Holdsworth MJ, Tucker GA, Knapp JE: Gene Expression during tomato ripening. Phil. Trans. R. Soc. Lond. B 314: 399–410 (1986).CrossRefGoogle Scholar
  11. 11.
    Grierson D, Tucker GA, Keen J, Ray J, Bird CR, Schuch W: Sequencing and identification of a cDNA clone for tomato polygalacturonase. Nucl. Acids. Res. 14: 8595–8603 (1986).PubMedCrossRefGoogle Scholar
  12. 12.
    Hall LN, Tucker GA, Smith CJS, Watson CF, Seymour GB, Bundick Y, Boniwell JM, Fletcher JD, Ray JA, Schuch W, Bird CR, Grierson D. Antisense inhibition of pectin esterase gene expression in transgenic tomatoes. The Plant Journal. 3: 121–129 (1993).CrossRefGoogle Scholar
  13. 13.
    Hamilton AJ, Lycett GW, Grierson D: Antisense gene that inhibits synthesis of the hormone ethylene in transgenic plants. Nature. 346: 284–287 (1990).CrossRefGoogle Scholar
  14. 14.
    Hamilton AJ, Bouzayen M, Grierson D: Identification of a tomato gene for the ethylene forming enzyme by expression in yeast. Proc. Natl. Acad. Sci. USA. 88: 7434–7437 (1991).PubMedCrossRefGoogle Scholar
  15. 15.
    Hobson GE: Polygalacturonase in normal and abnormal tomato fruit. Biochem. J. 92: 324–332 (1964).PubMedGoogle Scholar
  16. 16.
    Hobson GE: The firmmess of tomato fruit in relation to polygalacturonase activity. J. Hort. Sci. 40: 66–72 (1965).Google Scholar
  17. 17.
    Horsch RB, Fry JE, Hoffman NL, Eiochholtz D, Rogers SG, Frayley RT: A simple and general method for transferring genes into plants. Science. 227: 1229–1231 (1985).CrossRefGoogle Scholar
  18. 18.
    John P: How plant molecular biologists revealed a surprising relationships between two enzymes, which took an enzyme out of a membrane where it was not located, and put it into the soluble phase where it could be studied. Plant Mol. Biol. Rep. 9: 192–192 (1991).CrossRefGoogle Scholar
  19. 19.
    Jorgensen R: Altered gene expression in plants due to trans interactions between homologous genes. TIBTECH. 8: 340–344 (1990).Google Scholar
  20. 20.
    Klee HJ, Haiford MB, Kretzner KA, Barry GF, Kishmore GM: Control of Ethylene synthesis by expression of a bacterial enzyme in transgenic tomato plants. The Plant Cell. 3: 1187–1193 (1991).PubMedCrossRefGoogle Scholar
  21. 21.
    Lincoln JE, Cordes S, Read E, Fischer RL: Regulation of gene expression by ethylene during Lycopersicon esculentum (Tomato) fruit development. Proc. Natl. Acad. Sci. USA 84: 2793–2797 (1987).PubMedCrossRefGoogle Scholar
  22. 22.
    Mansson PE, Hsu D, Stalker D: Characterisation of fruit specific cDNAs from tomato. Mol. Gen. Genet. 200: 356–361 (1985).CrossRefGoogle Scholar
  23. 23.
    Maunders MJ, Holdsworth MJ, Slater A, Knapp JE, Bird CR, Schuch W, Grierson D: Ethylene stimulates the accumulation of ripening-related mRNAs in tomatoes. Plant Cell Envir. 10: 177–184 (1987).Google Scholar
  24. 24.
    McCormick S, Neidermeyer J, Fry J, Barnasson A, Horsch A, Frayley R: Leat disc transformation of cultivated tomato (L. esculentum) using Agrobacterium tumefaciens. Plant Cell Rep. 5: 81 (1986).CrossRefGoogle Scholar
  25. 25.
    Oeller PW, Wong LM, Taylor LP, Pike DA, Theologis A: Reversible inhibition of tomato fruit senescence by antisense 1-aminocyclopropane-1-carboxylate synthase. Science. 254: 437–439 (1991).PubMedCrossRefGoogle Scholar
  26. 26.
    Picton S, Barton SL, Bouzayen M, Hamilton AJ, Grierson D: Altered fruit ripening and leaf senescence in tomatoes expressing an antisense ethylene-forming enzyme transgene. The Plant Journal. 3: 469–481 (1993).CrossRefGoogle Scholar
  27. 27.
    Picton S, Gray J, AbuBaker U, Lowe A, Grierson D: cDNA cloning and characterisation of novel ripening-related mRNAs with altered patterns of accumulation in the ripening inhibitor (rin) tomato ripening mutant. Plant Mol.Biol. Submitted for publication (1993).Google Scholar
  28. 28.
    Ray J, Bird C, Maunders M, Grierson D, Schuch W: Sequence of pTOM 5, a ripening related cDNA from tomato. Nucl. Acids Res. 15: 10587 (1987).PubMedCrossRefGoogle Scholar
  29. 29.
    Redenbaugh K, Hiatt W, Martineau B, Kramer M, Sheehy R, Sanders R, Houck C, Emlay D: Safety Assessment of Genetically Engineered fruits and Vegetables. A Case study of the Flavr Savr™ Tomato. CRC Press, Boca Ratón, USA. (1992).Google Scholar
  30. 30.
    Sato T, Theologis A: Cloning the mRNA encoding 1-amino-cyclopropane-1-carboxylate synthase, the key enzyme for ethylene biosynthesis in plants. Proc. Natl. Acad. Sci. USA. 86: 6621–6625 (1989).PubMedCrossRefGoogle Scholar
  31. 31.
    Sato T, Oeller PW, Theologis A: The 1-aminocyclopropane-1-carboxylate synthase of Curcubita. J. Biol.Chem. 266: 3752–3759 (1991).PubMedGoogle Scholar
  32. 32.
    Schuch W, Hobson G, Kanczler J, Tucker G, Robertson D, Bright S, Bird C: Improvement of tomato fruit quality through genetic engineering. HortScience. 26: 1517–1520 (1991).Google Scholar
  33. 33.
    Sheehy RE, Pearson J, Brady CJ, Hiatt WR: Molecular characterization of tomato fruit polygalacturonase. Mol. Gen. Genet. 208: 30–36 (1987).CrossRefGoogle Scholar
  34. 34.
    Sheehy RE, Kramer M, Hiatt WR: Reduction of polygalacturonase activity in tomato fruit by antisense RNA. Proc. Natl. Acad. Sci. USA. 85: 8805–8809 (1988).PubMedCrossRefGoogle Scholar
  35. 35.
    Slater A, Maunders MJ, Edwards K, Schuch W, Grierson D: Isolation and characterization of cDNA clones for tomato polygalacturonase and other ripening related proteins. Plant Mol. Biol. 5: 137–147 (1985).CrossRefGoogle Scholar
  36. 36.
    Smith CJS, Slater A, Grierson D: Rapid appearance of mRNA correlated with ethylene synthesis encoding a protein of MW 35,000. Planta. 168: 94–100 (1986).CrossRefGoogle Scholar
  37. 37.
    Smith CJS, Watson CF, Ray J, Bird CR, Morris PC, Schuch W and Grierson D: Antisense RNA inhibition of polygalacturonase gene expression in transgenic tomatoes. Nature. 334: 724–726 (1988).CrossRefGoogle Scholar
  38. 38.
    Smith CJS, Watson CF, Bird CR, Ray J, Schuch W, Grierson D: Expression of a truncated tomato polygalacturonase gene inhibits expression of the endogenous gene in transgenic plants. Mol.Gen.Genet. 224: 477–481 (1990).PubMedCrossRefGoogle Scholar
  39. 39.
    Smith CJS, Watson CF, Morris PC, Bird CR, Seymour GB, Gray JE, Arnold C, Tucker GA, Schuch W, Harding S, Grierson D: Inheritance and effect on ripening of antisense polygalacturonase genes in transgenic tomatoes. Plant Mol.Biol. 14: 369–379 (1990).PubMedCrossRefGoogle Scholar
  40. 40.
    Spanu P, Reinhart D, Boller T: Analysis and cloning of the ethylene-forming enzyme from tomato by functional expression of its mRNA in Xenopus laevis oocytes. EMBO. J. 10: 2007–2013 (1991).PubMedGoogle Scholar
  41. 41.
    Themmen APN, Tucker GA, Grierson D: Degradation of isolated tomato cell walls by purified polygalacturonase in vitro. Plant Physiol. 69: 122–124 (1982).PubMedCrossRefGoogle Scholar
  42. 42.
    Tieman DM, Harriman RW, Ramamohan G, Handa AK: An antisense pectinmethylesterase gene alters pectin chemistry and soluble solids in tomato fruit. The Plant Cell. 4: 681–687 (1992).CrossRefGoogle Scholar
  43. 43.
    van der Krol AR, Lenting PE, Veenstra J, van der Meer IM, Koes RE, Gerats AGM, Mol JNM and Stuitje AR: An anti-sense chalcone synthase gene in transgenic plants inhibits flower pigmentation. Nature. 333: 866–869 (1988).CrossRefGoogle Scholar
  44. 44.
    Ververidis P, John P: Complete recovery in vitro of ethylene forming enzyme activity. Phytochemistry. 30: 725–727 (1991).CrossRefGoogle Scholar
  45. 45.
    Yang SF, Hoffmann NE: Ethylene biosynthesis and its regulation in higher plants. Ann. Rev. Plant Physiol. 35: 155–189 (1984).CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1994

Authors and Affiliations

  • Steve Picton
    • 1
  • Julie E. Gray
    • 1
  • Don Grierson
    • 1
  1. 1.Department of Physiology and Environmental Science, Sutton Bonington CampusUniversity of Nottingham, AFRC Research Group in Plant Gene RegulationLoughborough, LeicsUK

Personalised recommendations