Skip to main content
SpringerLink
Log in

Apple ACC-oxidase and polygalacturonase: ripening-specific gene expression and promoter analysis in transgenic tomato

  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

Abstract

Levels of 1-aminocyclopropane-1-carboxylate (ACC) oxidase and polygalacturonase (PG) mRNAs were characterized during ripening of Royal Gala, Braeburn and Granny Smith apples. Both ACC-oxidase and PG mRNAs were up-regulated in ripening fruit of all three cultivars. Expression in Royal Gala was detected earlier than in Braeburn and Granny Smith, relative to internal ethylene concentration. Genomic clones corresponding to the ACC-oxidase and PG mRNAs expressed in ripe apple fruit were isolated and ca. 2 kb of each promoter was sequenced. The start point of transcription in each gene was mapped by primer extension, and sequences homologous to elements in other ethylene-responsive or PG promoters were identified. The fruit specificity of the apple ACC-oxidase and PG promoters was investigated in transgenic tomato plants using a nested set of promoter fragments fused to the β-glucuronidase (gusA) reporter gene. For the ACC-oxidase gene, 450 bp of 5′ promoter sequence was sufficient to drive GUS expression, although this expression was not specific to ripening fruit. Larger fragments of 1966 and 1159 bp showed both fruit and ripening specificity. For the PG gene, promoter fragments of 1460 and 532 bp conferred ripening-specific expression in transgenic tomato fruit. However GUS expression was down-regulated by 2356 bp of promoter, suggesting the presence of a negative regulatory element between positions -1460 and -2356.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Atkinson RG: A cDNA clone for endopolygalacturonase from apple. Plant Physiol 105: 1437–1438 (1994).

    Article  PubMed  Google Scholar 

  2. Atkinson RG, Gardner RC: A polygalacturonase gene from kiwifruit (Actinidia deliciosa). Plant Physiol 103: 669–670 (1993).

    Article  PubMed  Google Scholar 

  3. Balagué C, Watson CF, Turner AJ, Rouge P, Picton S, Pech J-C, Grierson D: Isolation of a ripening and wound-induced cDNA from Cucumis melo L. encoding a protein with homology to the ethylene-forming enzyme. Eur J Biochem 212: 27–34 (1993).

    PubMed  Google Scholar 

  4. Bird CR, Smith CJS, Ray JA, Moureau 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).

    Google Scholar 

  5. Bolitho KM, Lay-Yee M, Knighton ML, Ross GS: Antisense apple ACC-oxidase RNA reduces ethylene production in transgenic tomato fruit. Plant Sci 122: 91–99 (1997).

    Article  Google Scholar 

  6. Brackman A, Streif J: Ethylene, CO2 and aroma volatiles production by apple cultivars. Acta Hort 368: 51–57 (1994).

    Google Scholar 

  7. Clarke HRG, Davis JM, Wilbert SM, Bradshaw HD Jr, Gordon MP: Wound-induced and developmental activation of a poplar tree chitinase gene promoter in transgenic tobacco. Plant Mol Biol: 25: 799–815 (1994).

    PubMed  Google Scholar 

  8. Deikman J, Fischer RL: Interaction of a DNA binding factor with the 5′-flanking region of an ethylene-responsive fruit ripening gene from tomato. EMBO J 7: 3315–3320 (1988).

    PubMed  Google Scholar 

  9. Deikman J, Kline R, Fischer RL: Organization of ripening and ethylene regulatory regions in a fruit-specific promoter from tomato (Lycopersicon esculentum). Plant Physiol 100: 2013–2017 (1992).

    Google Scholar 

  10. DellaPenna D, Alexander DC, Bennett AB: Molecular cloning of tomato fruit polygalacturonase: analysis of polygalacturonase mRNA levels during ripening. Proc Natl Acad Sci USA 83: 6420–6424 (1986).

    Google Scholar 

  11. Dellaporta SL, Wood J, Hicks JB: A plant DNA minipreparation: Version II. Plant Mol Biol Rep 1: 19–21 (1983).

    Google Scholar 

  12. Fischer RL, Bennett AB: Role of cell wall hydrolases in fruit ripening. Annu Rev Plant Physiol Plant Mol Biol 42: 675–703 (1991).

    Article  Google Scholar 

  13. Giovannoni JJ, DellaPenna D, Bennett 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).

    PubMed  Google Scholar 

  14. Gleave AP: A versatile binary vector system with a T-DNA organizational structure conducive to efficient integration of cloned DNA into the plant genome. Plant Mol Biol 20: 1203–1207 (1992).

    PubMed  Google Scholar 

  15. Grabowski PJ: Characterization of RNA. In: Higgins SJ, Hames BD (eds), RNA Processing: A Practical Approach. IRL Press, Oxford University, Oxford (1994).

    Google Scholar 

  16. Gussman CD, Goffreda JC, Gianfagna TJ: Ethylene production and fruit-softening rates in several apple fruit ripening variants. HortScience 28: 135–137 (1993).

    Google Scholar 

  17. Hamilton AJ, Lycett GW, Grierson D: Antisense gene that inhibits synthesis of the hormone ethylene in transgenic plants. Nature 346: 284–287 (1990).

    Article  Google Scholar 

  18. Holdsworth MJ, Bird CR, Ray J, Schuch W, Grierson D: Structure and expression of an ethylene-related mRNA from tomato. Nucl Acids Res 15: 731–739 (1987).

    PubMed  Google Scholar 

  19. Holdsworth MJ, Schuch W, Grierson D: Nucleotide sequence of an ethylene-related gene from tomato. Nucl Acids Res 15: 10600 (1987).

    PubMed  Google Scholar 

  20. Hollick JB, Gordon MP: A poplar tree proteinase inhibitor-like gene promoter is responsive to wounding in transgenic tobacco. Plant Mol Biol 22: 561–572 (1993).

    PubMed  Google Scholar 

  21. Itzhaki H, Maxson JM, Woodson WR: An ethylene-responsive enhancer element is involved in the senescence-related expression of the carnation glutathione-S-transferase (GST1) gene. Proc Natl Acad Sci USA 91: 8925–8929 (1994).

    PubMed  Google Scholar 

  22. Joshi CP: An inspection of the domain between putative TATA box and translation start site in 79 plant genes. Nucl Acids Res 15: 6644–6652 (1987).

    Google Scholar 

  23. Lasserre E, Bouquin T, Hernandez JA, Bull J, Pech J-C, Balagué C: Structure and expression of three genes encoding ACC oxidase homologs from melon (Cucumis melo L.) Mol Gen Genet 251: 81–90 (1996).

    Article  PubMed  Google Scholar 

  24. Lester DR, Speirs J, Orr G, Brady CJ: Peach (Prunus persica) endopolygalacturonase cDNA isolation and mRNA analysis in melting and nonmelting peach cultivars. Plant Physiol 105: 225–231 (1994).

    Article  PubMed  Google Scholar 

  25. Lin E, Burns DJW, Gardner RC: Fruit developmental regulation of the kiwifruit actinidin promoter is conserved in transgenic petunia plants. Plant Mol Biol 23: 489–499 (1993).

    Article  PubMed  Google Scholar 

  26. Lin HC, Lei SP, Wilcox G: An improved DNA sequencing strategy. Anal Biochem 147: 114–119 (1985).

    PubMed  Google Scholar 

  27. MacDiarmid CWB, Gardner RC: A cDNA sequence from kiwifruit homologous to 1-aminocyclopropane-1-carboxylic acid oxidase. Plant Physiol 101: 691–692 (1993).

    PubMed  Google Scholar 

  28. Montgomery J, Goldman S, Deikman J, Margossian L, Fischer RL: Identification of an ethylene-responsive region in the promoter of a fruit ripening gene. Proc Natl Acad Sci USA 90: 5939–5943 (1993).

    PubMed  Google Scholar 

  29. Montgomery J, Pollard V, Deikman J, Fischer RL: Positive and negative regulatory regions control the spatial distribution of polygalacturonase transcription in tomato fruit pericarp. Plant Cell 5: 1049–1062 (1993).

    PubMed  Google Scholar 

  30. Nicholass FJ, Smith CJS, Schuch W, Bird CR, Grierson D: High levels of ripening-specific reporter gene expression directed by tomato fruit polygalacturonase gene-flanking regions. Plant Mol Biol 28: 423–435 (1995).

    PubMed  Google Scholar 

  31. Ross GS, Knighton ML, Lay-Yee M: An ethylene-related cDNA from ripening apples. Plant Mol Biol 19: 231–238 (1992).

    PubMed  Google Scholar 

  32. Ross GS, Wegrzyn T, MacRae EA, Redgwell RJ: Apple β-galactosidase: activity against cell wall polysaccharides and characterization of a related cDNA clone. Plant Physiol 106: 521–528 (1994).

    PubMed  Google Scholar 

  33. Saltveit ME Jr: Procedures for extracting and analyzing internal gas samples from plant tissues by gas chromatography. HortScience 17: 878–881 (1982).

    Google Scholar 

  34. Sambrook J, Fritsch EF, Maniatis T: Molecular Cloning. A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1989).

    Google Scholar 

  35. Santino CG, Stanford GL, Conner TW: Developmental and transgenic analysis of two tomato fruit enhanced genes. Plant Mol Biol 33: 405–416 (1997).

    PubMed  Google Scholar 

  36. Schneider A, Salamini F, Gebhardt C: Expression patterns and promoter activity of the cold-regulated gene ci21A of potato. Plant Physiol 113: 335–345 (1997).

    PubMed  Google Scholar 

  37. Sheehy RE, Pearson J, Brady CJ, Hiatt WR: Molecular characterization of tomato fruit polygalacturonase. Mol Gen Genet 208: 30–36 (1987).

    Article  Google Scholar 

  38. Shinshi H, Usami S, Ohme-Takagi M: Identification of an ethylene-responsive region in the promoter of a tobacco class I chitinase gene. Plant Mol Biol 27: 923–932 (1995).

    PubMed  Google Scholar 

  39. Speirs J, Lee E, Brady CJ, Robertson J, McGlasson WB: En-dopolygalacturonase: Messenger RNA, enzyme and softening in the ripening fruit of a range of tomato genotypes. J Plant Physiol 135: 576–582 (1989).

    Google Scholar 

  40. van der Meer IM, Spelt CE, Mol JNM, Stuitje AR: Promoter analysis of the chalcone synthase (chsA) gene of Petunia hybrida: a 67 bp promoter region directs flower-specific expression. Plant Mol Biol 15: 95–109 (1990).

    Article  PubMed  Google Scholar 

  41. Van Haaren MJJ, Houck CM: Strong negative and positive regulatory elements contribute to the high-level fruit-specific expression of the tomato 2A11 gene. Plant Mol Biol 17: 615–630 (1991).

    PubMed  Google Scholar 

  42. Van Haaren MJJ, Houck CM: A functional map of the fruit-specific promoter of the tomato 2A11 gene. Plant Mol Biol 21: 625–640 (1993).

    PubMed  Google Scholar 

  43. Wu Q, Szakács-Dobozi M, Hemmat M, Hrazdina G: Endopolygalacturonase in apples (Malus domestica) and its expression during fruit ripening. Plant Physiol 102: 219–225 (1993).

    PubMed  Google Scholar 

  44. Yao J-L, Cohen D, Atkinson RG, and Morris BAM: Genetic transformation in apple (Malus × domestica). In: Bajaj YPS (ed) Biotechnology in Agriculture and Forestry: Plant Protoplasts and Genetic Engineering. Springer. Verlag Berlin/Heidelberg/New York (in press).

  45. Zhu J, Gardiner SE, Lay-Yee M: Physical mapping of three fruit ripening genes: endopolygalacturonase, ACC oxidase and ACC synthase from apple (Malus × domestica) in an apple rootstock A106 (Malus sieboldii). Cell Res 5: 243–253 (1995).

    Google Scholar 

Download references

Author information

Author notes

  1. Teresa Iturriagagoitia-Bueno

    Present address: Dept of Agricultural Botany, University of Reading, Whiteknights, P.O. Box 227, Reading, RG6 2AS, UK

Authors and Affiliations

  1. Horticultural Research Institute of New Zealand, Mt Albert Research Centre, Private Bag 92169, Auckland, New Zealand

    Ross G. Atkinson, Karen M. Bolitho, Michele A. Wright, Suzanne J. Reid & Gavin S. Ross

Authors
  1. Ross G. Atkinson
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Karen M. Bolitho
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Michele A. Wright
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Teresa Iturriagagoitia-Bueno
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Suzanne J. Reid
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Gavin S. Ross
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Atkinson, R.G., Bolitho, K.M., Wright, M.A. et al. Apple ACC-oxidase and polygalacturonase: ripening-specific gene expression and promoter analysis in transgenic tomato. Plant Mol Biol 38, 449–460 (1998). https://doi.org/10.1023/A:1006065926397

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1006065926397

Search

Navigation