Skip to main content
SpringerLink
Log in

Isolation and characterisation of a pod dehiscence zone-specific polygalacturonase fromBrassica napus

  • Regular Article
  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

Abstract

Seven distinct partial cDNAs, similar in sequence to previously described polygalacturonases (PGs), were amplified from cDNA derived from rape pod wall, dehiscence zone and leaves by the polymerase chain reaction. Northern analysis showed that one clone, PG35-8, was expressed at low levels in the dehiscence zone during the first five weeks after anthesis but was very abundantly expressed at week 6. In contrast, no PG35-8-related RNA was detected in the pod wall. Our data suggest that there are temporal and spatial correlations between the breakdown of the middle lamella, of the dehiscence zone cells and the pattern of synthesis of PG35-8 transcripts which may indicate a role for this particular PG in rape pod dehiscence.

PG35-8 was used to isolate five cDNA clones from a rape dehiscence zone cDNA library. Restriction enzyme analysis and partial sequencing revealed that they were derived from four highly homologous transcripts which are probably allelic forms of a single gene. One full-length clone, RDPG1, was completely sequenced. The predicted protein of RDPG1 showed its highest identity with PG from apple fruit with an identity of 52%.

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

    Article  PubMed  Google Scholar 

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

    Article  PubMed  Google Scholar 

  3. Baird LM, Reid PD: Cellulase activity and localization during induced abscission ofColeus blumei. J Plant Growth Regul 11: 129–134 (1992).

    Article  Google Scholar 

  4. Blumenkrantz N, Asboe-Hansen G: New method for quantitative determination of uronic acids. Anal Biochem 54: 484–489 (1973).

    PubMed  Google Scholar 

  5. Bonghi C, Rascio N, Ramina A, Casadoro G: Cellulase and polygalacturonase involvement in the abscission of leaf and fruit explants of peach. Plant Mol Biol. 20: 839–848 (1992).

    PubMed  Google Scholar 

  6. Brown SM, Crouch ML: Characterization of a gene family abundantly expressed inOenothera organensis pollen that shows sequence similarity to polygalacturonase. Plant Cell 2: 263–274 (1990).

    Article  PubMed  Google Scholar 

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

    Google Scholar 

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

    Google Scholar 

  9. Devereux J, Haeberli P, Smithies O: A comprehensive set of sequence software programs for VAX. Nucl Acids Res 12: 387–395 (1984).

    PubMed  Google Scholar 

  10. Dopico B, Lowe AL, Wilson ID, Merodio C, Grierson D: Cloning and characterization of avocado fruit mRNAs and their expression during ripening and low-temperature storage. Plant Mol Biol 21: 437–449 (1993).

    Article  PubMed  Google Scholar 

  11. del Campillo E, Reid PD, Sexton R, Lewis LN: Occurrence and localization of 9.5 cellulase in abscising and nonabscising tissues. Plant Cell 2: 245–254 (1990).

    Article  PubMed  Google Scholar 

  12. Feinberg AP, Vogelstein B: A technique for radio-labeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132: 6–13 (1983).

    PubMed  Google Scholar 

  13. 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 

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

    PubMed  Google Scholar 

  15. John EM, Petersen MW: Cotton (Gossypium hirsutum L) pollen-specific polygalacturonase mRNA: tissue and temporal specificity of its promotor in transgenic tobacco. Plant Mol Biol 26: 1989–1993 (1994).

    PubMed  Google Scholar 

  16. Josefson E: Investigation into shattering resistance of cruciferous oil crops. Zeitschr Pflanzenzucht 59: 384–396 (1968).

    Google Scholar 

  17. Kalaitzis P, Koehler SM, Tucker ML: Cloning of a tomato polygalacturonase expressed in abscission. Plant Mol Biol 28: 647–656 (1995).

    PubMed  Google Scholar 

  18. Kutsunai SY, Lin AC; Percival FW, Laties GG, Christoffersen RE: Ripening-related polygalacturonase cDNA from avocado. Plant Physiol 103: 289–290 (1993).

    Article  PubMed  Google Scholar 

  19. Lashbrook CC, Gonzales-Bosch C, Bennett AB: Two divergent endo-b-1,4-glucanase genes exhibit overlapping expression in ripening fruit and abscising flowers. Plant Cell 6: 1485–1493 (1994).

    Article  PubMed  Google Scholar 

  20. 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 

  21. Meakin PJ, Roberts JA. Dehiscence of fruit in oilseed rape (Brassica napus L) I. Anatomy of pod dehiscence. J Exp Bot 229: 995–1002 (1990).

    Google Scholar 

  22. Meakin PJ, Roberts JA. Dehiscence of fruit in oilseed rape (Brassica napus L) II. The role of cell wall degrading enzymes and ethylene. J Exp Bot 229: 1003–1011 (1990).

    Google Scholar 

  23. Niogret MF, Dubald M, Mandaron P, Mache R: Characterization of pollen polygalacturonase encoded by several cDNA clones in maize. Plant Mol Biol 17: 1155–1167 (1991).

    PubMed  Google Scholar 

  24. Orr G, Brady C: Relationship of endopolygalacturonase activity to fruit softening in a freestone peach. Postharv Biol Technol 3: 121–130 (1993).

    Article  Google Scholar 

  25. Osborne DJ: Abscission. CRC Crit Rev Plant Sci 2: 103–129 (1989).

    Google Scholar 

  26. Picart JA, Morgan DG: Preliminary studies on the effect of 4-chlorophenoxyacetic acid on the senescence and dehiscence of pods in oil seed rape (Brassica napus). Plant Growth Regul 4: 169–175 (1986).

    Google Scholar 

  27. Robert LS, Allard S, Gerster JL, Cass L, Simmonds J: Isolation and characterization of a polygalacturonase gene highly expressed in Brassica napus pollen. Plant Mol Biol 23: 1273–1278 (1993).

    PubMed  Google Scholar 

  28. Rogers HJ, Allen RL, Hamilton WDO and Lonsdale DM: Pollen specific cDNA clone fromZea mays. Biochim Biophys Acta 1089: 411–413 (1991).

    PubMed  Google Scholar 

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

    Google Scholar 

  30. Sexton R, Jamieson GGC, Hall MHIL: An ultrastructual study of abscission zone cells with special reference to the mechanism of enzyme secretion. Protoplasma 91: 369–387 (1977).

    Google Scholar 

  31. Smith CJS, Watson CF, Ray J, Bird CR, Morris PC, Schuch W, Grierson D: Antisense RNA inhibition of polygalacturonase gene expression in ripening tomatoes. Nature 334: 724–726 (1988).

    Article  Google Scholar 

  32. Taylor JE, Tucker GA, Lasslett Y, Smith CJS, Arnold CM, Watson CF, Schuch W, Grierson D, Roberts JA: Polygalacturonase expression during leaf abscission of normal and transgenic tomato plants. Planta 133–138 (1990).

  33. Tebbutt SJ, Rogers HJ, Lonsdale DM: Characterization of a tobacco gene encoding a pollen-specific polygalacturonase. Plant Mol Biol 25: 283–297 (1994).

    Article  PubMed  Google Scholar 

  34. Wegrzyn TF, MacRae EA: Pectinesterase, polygalacturonase and β-galactosidase during softening of ethylene-treated kiwifruit. Hort Science 27: 900–902 (1992).

    Google Scholar 

  35. Wu Q, Szakacs-Dobozi M, Hennat M, Hrazdina G: Endo-poly-galacturonase in apples (Malus domestica) and its expression during fruit ripening. Plant Physiol 102: 219–225 (1993).

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. The Biotechnology Group, DIPS, DK-2800, Lottenborgvej 2, Lyngby

    Morten Petersen, Lilli Sander, Peter Ulvskov & Bernhard Borkhardt

  2. IACR-Long Ashton Research Station, Department of Agricultural Sciences, University of Bristol, BS18 9AF, Long Ashton, UK

    Robin Child

  3. Department of Biochemistry, University of Antwerp, 1 Universiteitsplein, Wilrijk, B-26 10, Antwerp, Belgium

    Harry van Onckelen

Authors
  1. Morten Petersen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Lilli Sander
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Robin Child
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Harry van Onckelen
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Peter Ulvskov
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Bernhard Borkhardt
    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

Petersen, M., Sander, L., Child, R. et al. Isolation and characterisation of a pod dehiscence zone-specific polygalacturonase fromBrassica napus . Plant Mol Biol 31, 517–527 (1996). https://doi.org/10.1007/BF00042225

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00042225

Key words

Search

Navigation