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The manipulation and modification of tomato fruit ripening by expression of antisense RNA in transgenic plants

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The Methodology of Plant Genetic Manipulation: Criteria for Decision Making

Part of the book series: Developments in Plant Breeding ((DIPB,volume 3))

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Summary

The common cultivated tomato (Lycopersicon esculentum Mill.) provides a major focus for improvement of crop quality through genetic engineering. Identification of ripening-related cDNAs has enabled the modification of specific aspects of ripening by manipulating gene expression in transgenic plants. By utilizing ‘antisense RNA’ to modify expression of ripening genes, we have inhibited the production of the cell wall - metabolising enzymes polygalacturonase and pectinesterase and created transgenic plants that contain, effectively, single, targeted mutations affecting these genes. Furthermore, this approach has been used with previously unidentified cDNA clones to enable both functional identification and manipulation of genes involved in ethylene production (ACC oxidase) and carotenoid biosynthesis (phytoene synthase). The use of antisense RNA targeted to specific genes to alter ripening phenotypes and improve commercial utility of fruit by affecting shelf-life, processing characteristics and nutritional content is discussed.

We have used the extreme ripening-impaired mutant, ripening inhibitor (rin) to identify additional genes implicated in the ripening process. This approach has resulted in the cloning of several novel ripening-related mRNAs which are now being studied by antisense experiments. This may enable identification and manipulation of additional genes involved in processes such as softening, flavour and aroma generation and susceptibility to pathogens.

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Abbreviations

ACC:

1-aminocyclopropane-1-carboxylic acid

PE:

pectinesterase

PG:

polygalacturonase

SAM:

S-adenosyl methionine

SARs:

scaffold attachment regions

References

  • Allen, G.C., G.E. Hall, L.C. Childs,A.K. Wiessinger, S. Spiker & W.F. Thompson, 1993. Scaffold attachmentregionsincrease reporter gene expression in stably transformed plant cells. Plant Cell5: 603–613.

    PubMed  CAS  Google Scholar 

  • Armstrong, G.A.,M. Alberti & J.E. Hearst, 1990. Conserved enzymes mediate the earlyreactions of carotenoid biosynthesis in non-photosynthetic and photosyntheticprokaryotes. Proc. Natl. Acad. Sci. USA 87: 9975–9979.

    Article  PubMed  CAS  Google Scholar 

  • Bird, C.R.,C.J.S. Smith, J.A. Ray, P. Moureau, M.W. Bevan, A.S. Bird, S. Hughes, P.C.Morris, D. Grierson & W. Schuch, 1988. The tomato polygalacturonase geneand ripening specific expression in transgenic plants. Plant Mol. Biol. 11:651–662.

    Article  CAS  Google Scholar 

  • Bird, C.R., J.A.Ray, J.D. Fletcher, J.M. Boniwell, A.S. Bird, C. Teulieres, I. Blain, P.M.Bramley & W. Schuch, 1991. Using antisense RNA to study genefunction: Inhibition of carotenoid biosynthesis in transgenic tomatoes.Bio-Technology 9: 635–639.

    Article  CAS  Google Scholar 

  • Bouzayen, M., A.Hamilton, S. Picton, S. Barton & D. Grierson, 1992. Identification of genesfor the ethylene-forming enzyme and inhibition of ethylene synthesis intransgenic plants using antisense genes. Biochem. Soc. Trans. 20: 76–79.

    PubMed  CAS  Google Scholar 

  • Breyne, P., M.Van Montague, A. Depiicker & G. Gheysen, 1992. Characterisation of a plantscaffold attachment region in a DNA fragment that normalizes transgeneexpression in tobacco. Plant Cell 4: 463–471.

    PubMed  CAS  Google Scholar 

  • Crookes, P.R.& D. Grierson, 1983. Ultrastructure of tomato fruit ripening and the roleof polygalacturonase isoenzymes in cell wall degradation. Plant Physiol. 72:1088–1093.

    Article  PubMed  CAS  Google Scholar 

  • Davies, K.M.& D. Grierson, 1989. Identification of cDNA clones for tomato(Lycopersicon esculentum) mRNAs that accumulate during fruit ripening andleaf senescence in response to ethylene. Planta 179: 73–80.

    Article  CAS  Google Scholar 

  • DellaPenna, D.,D.C. Alexander & A.B. Bennett, 1986. Molecular cloning of tomato fruitpolygalacturonase: analysis of polygalacturonase mRNA levels during ripening.Proc. Natl. Acad. Sci.USA83: 6420–6424.

    Article  PubMed  CAS  Google Scholar 

  • Fray, R.G. &D. Grierson, 1993. Identification and genetic analysis of normal and mutantphytoene synthase genes of tomato by sequencing, complementation andco-suppression. Plant Mol. Biol. 22: 589–602.

    Article  PubMed  CAS  Google Scholar 

  • Giovannoni, J.J.,D. DellaPenna, A.B. Bennett & R.L. Fischer, 1989. Expression of a chimericpolygalacturonase gene in transgenicrin (ripening inhibitor) tomatofruit results in polyuronide degradation but not fruit softening. Plant Cell1: 53–63.

    PubMed  CAS  Google Scholar 

  • Gray, J.E., S.Picton, J. Shabbeer, W. Schuch & D. Grierson, 1992. Molecular biology offruit ripening and its manipulation with antisense genes. Plant Mol. Biol. 19: 69–87.

    Article  PubMed  CAS  Google Scholar 

  • Gray, J.E., S.Picton & D. Grierson, 1994. The use of transgenic and naturally occurringmutants to understand and manipulate tomato fruit ripening. Plant, Cell &Environ. 17: 557–571.

    Article  CAS  Google Scholar 

  • Grierson, D.,M.J. Maunders, A. Slater, J. Ray, C.R. Bird, W. Schuch, M.J. Holdsworth, G.A.Tucker & J.E. Knapp,1986a.Gene expression during tomato ripening. Phil. Trans. R. Soc. Lond. B 314:399–410.

    Article  CAS  Google Scholar 

  • Grierson, D.,G.A. Tucker, J. Keen, J. Ray, C.R. Bird & W. Schuch, 1986b. Sequencing andidentification of a cDNA clone for tomato polygalacturonase. Nucl. Acids Res.14: 8595–8603.

    Article  PubMed  CAS  Google Scholar 

  • Grierson, D.,M.E. Purton,J.E.Knapp&B. Bathgate, 1987. Tomato ripening mutants. In: H. Thomas & D. Grierson(Eds) Developmental Mutants in Higher Plants, pp. 73–94. Cambridge UniversityPress, Cambridge.

    Google Scholar 

  • Grierson, D.& W. Schuch, 1993. Control of ripening. Phil. Trans. R. Soc. Lond. B342,241–250.

    Google Scholar 

  • Hall, L.N., G.A.Tucker, C.J.S. Smith, C.F. Watson, G.B. Seymour, Y. Bundick, J.M. Boniwell,J.D. Fletcher, J.A. Ray, W. Schuch, C.R. Bird & D. Grierson, 1993.Antisense inhibition of pectinesterase gene expression in transgenic tomatoes.The Plant Journal 3: 121–129.

    Article  CAS  Google Scholar 

  • Hamilton, A.J., G.W. Lycett & D.Grierson, 1990. Antisense gene that inhibits synthesis of the hormone ethylenein transgenic plants. Nature 346: 284–287.

    Article  CAS  Google Scholar 

  • Hamilton, A.J.,M. Bouzayen & D. Grierson, 1991. Identification of a tomato gene for theethylene forming enzyme by expression in yeast. Proc. Natl. Acad. Sci. USA 88:7434–7437.

    Article  PubMed  CAS  Google Scholar 

  • Hobson, G.E.,1965. The firmness of tomato fruit in relation to polygalacturonase activity.J. Hort.Sci. 40:66–72.

    CAS  Google Scholar 

  • Holdsworth, M.J.,C.R. Bird, J. Ray, W. Schuch & D. Grierson, 1987. Structure and expressionof an ethylene-related mRNA from tomato. Nucl. Acids.Res. 15: 731–739.

    Article  PubMed  CAS  Google Scholar 

  • Holdsworth, M.J.,W. Schuch & D. Grierson, 1988. Organisation and expression of awound/ripening-related small multigene family from tomato. Plant Mol. Biol. 11: 81–88.

    Article  CAS  Google Scholar 

  • John, P., 1991.How plant molecular biologists revealed a surprising relationship between twoenzymes, which took an enzyme out of a membrane where it was not located, andput it into the soluble phase where it could be studied. Plant Mol. Biol. Rep. 9: 192–194.

    Article  Google Scholar 

  • Kramer, M., R.A.Sanders, R.E. Sheehy, M. Melis, M. Kuchn & W.R. Hiatt, 1990. Fieldevaluation of tomatoes with reduced polygalacturonase by antisense RNA. In: A.B. Bennett& S.D. O’Niel (Eds) Horticultural Biotechnology, pp. 347–355. Wiley- Liss,New York.

    Google Scholar 

  • Kramer, M., R.Sanders, H. Bolkan, C. Waters, R.E. Sheehy & R.W. Hiatt, 1992. Postharvestevaluation of transgenic tomatoes with reduced levels of polygalacturonase:Processing, firmness and disease resistance. Postharvest Biol. Technol. 1:241–255.

    Article  CAS  Google Scholar 

  • Lincoln, J.E., S.Cordes,E.Read & R.L. Fischer, 1987. Regulation of gene expression by ethylene duringLycopersicon esculentum (Tomato) fruit development. Proc. Natl. Acad. Sci. USA 84:2793–2797.

    Article  PubMed  CAS  Google Scholar 

  • Murray, A.J.,G.E. Hobson, W. Schuch & C.R. Bird, 1993. Reduced ethylene biosynthesis inEFE-antisense tomatoes has differential effects on the ripening process.Postharvest Biol. Technol. 2: 301–313.

    Article  CAS  Google Scholar 

  • Nicholass, F.J.,C.F. Watson, C.J.S. Smith, W. Schuch, C.R. Bird & D. Grierson, 1995. Highlevels of ripening-specific reporter gene expression directed by tomato fruitpolygalacturonase gene flanking regions. Plant Mol. Biol., in press.

    Google Scholar 

  • Oeller, P.W., L.M. Wong,L.P. Taylor, D.A. Pike & A. Theolo- gis, 1992. Reversible inhibition oftomato fruit senescence by antisense 1-aminocyclopropane-1-carboxylatesynthase. Science 254: 427–439.

    Google Scholar 

  • Peach, C. &J. Velten,1991.Transgeneexpressionvariability (position effect) of CAT and GUS reporter genesdriven by linked divergent T-DNA promoters. Plant Mol. Biol. 17: 49–60.

    Article  PubMed  CAS  Google Scholar 

  • Peñarrubia, L.,M. Aguilar, L. Margossian & R.L. Fischer, 1992. An antisense genestimulates ethylene hormone production during tomato fruit ripening. Plant Cell4: 681–687.

    PubMed  Google Scholar 

  • Picton, S., S.L.Barton, M. Bouzayen, A.J. Hamilton & D. Grierson, 1993a. Altered fruitripening and leaf senescence in tomatoes expressing an antisenseethylene-forming enzyme transgene. The Plant Journal 3: 469–481.

    Article  CAS  Google Scholar 

  • Picton, S., J.E.Gray, S. Barton, U. AbuBaker, A. Lowe & D. Grierson, 1993b. cDNA cloningand characterisation of novel ripening-related mRNAs with altered patterns ofaccumulation in theripening inhibitor (rin) tomato ripening mutant.Plant Mol.Biol.23: 193–207.

    Article  PubMed  CAS  Google Scholar 

  • Picton, S., J.E.Gray, S. Payton, S. Barton, A. Lowe & D. Grierson, 1993c. A histidinedecarboxylase-like mRNA is involved in tomato fruit ripening. Plant Mol. Biol. 23:627–631.

    Article  PubMed  CAS  Google Scholar 

  • Picton, S., J.E.Gray & D. Grierson, 1994. Ethylene genes and fruit ripening. In: P.J.Davies (Ed) Plant Hormones: Physiology, Biochemistry and Molecular Biology, inpress. Kluwer Academic Publishers, The Netherlands.

    Google Scholar 

  • Ray, J., C. Bird,M. Maunders, D. Grierson & W. Schuch, 1987. Sequence of pTOM 5, a ripeningrelated cDNA from tomato. Nucl. Acids Res. 15: 10587.

    Article  PubMed  CAS  Google Scholar 

  • Redenbaugh, K.,W. Hiatt, B. Martineau, M. Kramer, R. Sheehy, R. Sanders, C. Houck & D.Emlay, 1991. Safety assessment of genetically engineered fruits and vegetables.A case study of the Flavr Savr™ tomato. CRC Press, Boca Raton, USA.

    Google Scholar 

  • Schuch, W., G.Hobson, K. Kanczler, G. Tucker, D. Robertson, D. Grierson, S. Bright & C.Bird, 1991. Improvement of tomato fruit quality through genetic engineering.HortScience 26: 1517–1520.

    CAS  Google Scholar 

  • Seymour, G.B.,R.G. Fray, P. Hill & G.A. Tucker, 1993. Down- regulation of twonon-homologous endogenous tomato genes with a single chimeric sense geneconstruct. Plant Mol.Biol. 23: 1–9.

    Article  PubMed  CAS  Google Scholar 

  • Sheehy, R.E., J.Pearson, C.J. Brady & W.R. Hiatt, 1987. Molecular characterization oftomato fruit polygalacturonase. Mol. Gen. Genet. 208: 30–36.

    Article  CAS  Google Scholar 

  • Sheehy, R.E., M.Crammer & W.R. Hiatt, 1988. Reduction of polygalacturonase activity intomato fruit by antisense RNA. Proc. Natl. Acad. Sci. USA 85:8805–8809.

    Article  PubMed  CAS  Google Scholar 

  • Slater, A., M.J.Maunders, K. Edwards, W. Schuch & D. Grierson, 1985. Isolation andcharacterization of cDNA clones for tomato polygalacturonase and other ripeningrelated proteins. Plant Mol. Biol. 5: 137–147.

    Article  CAS  Google Scholar 

  • Smith, C.J.S., A.Slater & D. Grierson, 1986. Rapid appearance of a mRNA correlated withethylene synthesis encoding a protein of MW 35,000. Planta 168: 94–100.

    Article  CAS  Google Scholar 

  • Smith, C.J.S.,C.F. Watson, J. Ray,C.R.Bird, P.C. Morris, W. Schuch & D. Grierson, 1988. Antisense RNA inhibition ofpolygalacturonase gene expression in transgenic tomatoes. Nature 334: 724–726.

    Article  CAS  Google Scholar 

  • Smith, C.J.S.,C.F. Watson, P.C. Morris, C.R. Bird, G.B. Seymour, J.E. Gray, C. Arnold, G.A.Tucker, W. Schuch, S. Harding & D. Grierson, 1990. Inheritance and effecton ripening of antisense polygalacturonase genes in transgenic tomatoes. Plant Mol. Biol. 14:369–379.

    Article  PubMed  CAS  Google Scholar 

  • Spanu, P., D. Reinhart & T. Boller,1991. Analysis and cloning of the ethylene-forming enzyme from tomato byfunctional expression of its mRNA inXenopus laevis oocytes. EMBO J. 10:2007–2013.

    PubMed  CAS  Google Scholar 

  • Tieman, D.M.,R.W. Harriman,G. Ramamohan & A.K. Handa, 1992. An antisense pectinmethylesterase genealters pectin chemistry and soluble solids in tomato fruit. Plant Cell 4:667–679.

    PubMed  CAS  Google Scholar 

  • van der Krol, A.R., P.E.Lenting, J.Veenstra, I.M. Van der Meer, R.E. Koes, A.G.M. Gerats, J.N.M. Mol & A.R. Stuitje, 1988. An anti-sense chalcone synthase gene in transgenic plantsinhibits flower pigmentation. Nature 333: 866–869.

    Article  Google Scholar 

  • Yang, S.F. &N.E. Hoffman, 1984. Ethylene biosynthesis and its regulation in higher plants.Ann. Rev. Plant Physiol. 35: 155– 189.

    Article  CAS  Google Scholar 

  • Yang, S.F., 1985.Biosynthesis and action of ethylene. HortScience 20: 41–45.

    CAS  Google Scholar 

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Author notes

  1. Steve Picton

    Present address: Kelvin close, Applied Biosystems Ltd., Birchwood Science Park North, Warrington, Cheshire, WA3 7PB, UK

Authors and Affiliations

  1. BBSRC Research Group in Plant Gene Regulation, Department of Physiology and Environmental Science, University of Nottingham, Sutton Bonington Campus, Loughborough, Leics., LE12 5RD, UK

    Steve Picton & Don Grierson

  2. Department of Molecular Biology and Biotechnology, Firth court, University of Sheffield, Western Bank, Sheffield, S10 2TN, UK

    Julie E. Gray

Authors
  1. Steve Picton
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  2. Julie E. Gray
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  3. Don Grierson
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Editors and Affiliations

  1. Department of Plant Science, University College, Cork, Ireland

    Alan C. Cassells  & Peter W. Jones  & 

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© 1995 Springer Science+Business Media New York

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Picton, S., Gray, J.E., Grierson, D. (1995). The manipulation and modification of tomato fruit ripening by expression of antisense RNA in transgenic plants. In: Cassells, A.C., Jones, P.W. (eds) The Methodology of Plant Genetic Manipulation: Criteria for Decision Making. Developments in Plant Breeding, vol 3. Springer, Dordrecht. https://doi.org/10.1007/978-94-011-0357-2_24

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  • DOI: https://doi.org/10.1007/978-94-011-0357-2_24

  • Publisher Name: Springer, Dordrecht

  • Print ISBN: 978-94-010-4159-1

  • Online ISBN: 978-94-011-0357-2

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