Plant Molecular Biology

, Volume 55, Issue 3, pp 369–388 | Cite as

Characterisation of Mal d 1-related genes in Malus

  • Lesley Beuning
  • Judith Bowen
  • Helena Persson
  • Diane Barraclough
  • Sean Bulley
  • Elspeth MacRae
Article

Abstract

It has been suggested that there are at least 15 Mal d 1-related (PR10) genes in one genotype of apple (Malus × domestica Borkh.). We sequenced cDNA libraries of cultivar ‘Royal Gala’ and identified 12 members of the Mal d 1 family, including the previously reported Mal d 1b and Mal d 1d, an allelic variant of the previously reported Mal d 1a. Eight Mal d 1 gene products were expressed in tree-ripened fruit, in either the cortex or the skin, and most of these were also expressed in leaves in response to challenge with Venturia inaequalis—a fungal disease of apple. Mal d 1 gene products were identified from a large number of different tissues. Degree of ripeness as measured by standard parameters was shown not to predict either the amount of protein able to bind to a specific monoclonal antibody 5H8, previously shown to bind to an allergenic epitope in Mal d 1b and a/d, or the amount of Mal d 1 mRNA present. Mal d 1d and Mal d 1b were the most highly expressed isoforms in ‘Royal Gala’, particularly in the skin of fruit, and these isoforms were also predominant in other cultivars and species of apple. Genotypes, however, differed in relative predominance of Mal d 1b and Mal d 1d. The predominantly expressed Mal d 1 genes in ripe apple fruit were translated in vivo into proteins and proteins binding to the antibody were found in all cultivars and species examined. New Mal d 1 proteins were identified that bound to the 5H8 antibody. At least two new subfamilies have been identified, and while some structural differences are predicted between groups of isoforms, the P-loop motif is identical in all except two isoforms. A role in intracellular signalling in plants is suggested and in vitro expression of the isoforms should help in assessing their relative roles in disease, allergic responses, senescence and nucleotide-, cytokinin- and brassinosteroid-binding.

allergen apple Mal d 1 PR10 protein 

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References

  1. Akkerdaas, J. H., van Ree, R., Aalbers, M., Stapel, S. O. and Aalberse, R. C. 1995. Multiplicity of cross-reactive epitopes on Bet v I as detected with monoclonal antibodies and human IgE. Allergy 50: 215-220.PubMedGoogle Scholar
  2. Altschul, S. F., Madden, T. L., Schaffer, A. A., Zhang, J., Zhang, Z., Miller, W. and Lipman, D. J. 1997. Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res. 25: 3389-3402.PubMedGoogle Scholar
  3. Atkinson, R. G., Perry, J., Matsui, T., Ross, G. S. and MacRae, E. A. 1996. A stress-, pathogenesis-, and allergen-related cDNA in apple fruit is also ripening-related. NZ J. Crop Hort. Sci. 24: 103-107.Google Scholar
  4. Bais, H. P., Vepachedu, R., Lawrence, C. B., Stermitz, F. R., Vivanco, J. M. 2003. Molecular and biochemical characterization of an enzyme responsible for the formation of hypericin in St John's wort (Hypericum perforatum L. ). J. Biol. Chem. 278: 32413-32422.PubMedGoogle Scholar
  5. Bantignies, B., Seguin, J., Muzac, I., Dedaldechamp, F., Gulick, P. and Ibrahim, R. 2000. Direct evidence for ribonucleolytic activity of a PR-10-like protein from white lupin roots. Plant Mol. Biol. 42: 871-881.PubMedGoogle Scholar
  6. Barraclough D., Obenland, D., Laing, W., Carroll, T. 2004. A general method for two-dimensional protein electrophoresis of fruit samples. Postharvest Biol. Technol. (in press)Google Scholar
  7. Breiteneder, H., Pettenburger, K., Bito, A., Valenta, R., Kraft, D., Rumpold, H., Scheiner, O. and Breitenbach, M. 1989. The gene coding for the major birch pollen allergen, Betv1, is highly homologous to a pea disease resistance response gene. EMBO J. 8: 1935-1938.PubMedGoogle Scholar
  8. Bufe, A., Spangfort, M. D., Kahlert, H., Schlaak, M. and Becker, W.-M. 1996. The major birch pollen allergen, Bet v 1, shows ribonuclease activity. Planta 199: 413-415.PubMedGoogle Scholar
  9. Chang, S., Puryear, J. and Cairney, J. 1993. A simple and efficient method for isolating RNA from pine trees. Plant Mol. Biol. Rep. 11: 113-116.Google Scholar
  10. Dreborg, S. 1988. Food allergy in pollen-sensitive patients. Ann. Allergy 61: 41-46.PubMedGoogle Scholar
  11. Ebner, C., Birkner, T., Valenta, R., Rumpold, H., Breitenbach, M., Scheiner, O. and Kraft, D. 1991. Common epitopes of birch pollen and apples-studies by western and northern blot. J. Allergy Clin. Immunol. 88: 588-594.PubMedGoogle Scholar
  12. Ebner, C., Hirschwehr, R., Bauer, L., Breiteneder, H., Valenta, R., Hoffmann, K., Krebitz, M., Kraft, D. and Scheiner, O. 1996. Identification of allergens in apple, pear, celery, carrot and potato: cross-reactivity with pollen allergens. Monogr. Allergy 32: 73-77.PubMedGoogle Scholar
  13. Ekramoddoullah, A. K. M., Yu, X., Sturrock, R., Zamani, A. and Taylor, D. 2000. Detection and seasonal expression pattern of a pathogenesis-related protein (PR-10) in Douglas-fir (Pseudotsuga menziesii) tissues. Physiol. Plant. 110: 240-247.Google Scholar
  14. Fahlbusch, B., Rudeschko, O., Müller, W. D., Schlenvoigt, G., Vettermann, S. and Jäger, L. 1995. Purification and characterization of the major allergen from apple and its allergenic cross-reactivity with Bet v 1. Int. Arch. Allergy Immunol. 108: 119-126.PubMedGoogle Scholar
  15. Fernández-Rivas, M. and Cuevas, M. 1999. Peels of Rosaceae fruits have a higher allergenicity than pulps. Clin. Exp. Allergy 29: 1239-1247.PubMedGoogle Scholar
  16. Ferreira, F., Hirtenlehner, K., Jilek, A., Godnik-Cvar, J., Breiteneder, H., Grimm, R., Hoffmann-Sommergruber, K., Scheiner, O., Kraft, D., Breitenbach, M., Rheinberger, H.-J. and Ebner, C. 1996. Dissection of immunoglobulin E and T lymphocyte reactivity of isoforms of the major birch pollen allergen Bet v1: potential use of hypoallergenic isoforms for immunotherapy. J. Exp. Med. 183: 599-609.PubMedGoogle Scholar
  17. Ferreira, F., Ebner, C., Kramer, B., Casari, G., Briza, P., Kungl, A. J., Grimm, R., Jahn-Schmid, B., Breiteneder, H., Kraft, D., Breitenbach, M., Rheinberger, H.-J. and Scheiner, O. 1998. Modulation of IgE reactivity of allergens by site-directed mutagenesis: potential use of hypoallergenic variants for immunotherapy. FASEB J. 12: 231242.Google Scholar
  18. Fujimoto, Y., Nagata, R., Fukasawa, H., Yano, K., Azuma, M., Iida, A., Sugimoto, S., Shudo, K. and Hashimoto, Y. 1998. Purification and cDNA cloning of cytokinin-specific binding protein from mung bean (Vigna radiata). Eur. J. Biochem. 258: 794-802.PubMedGoogle Scholar
  19. Gajhede, M., Osmark, P., Poulsen, F. M., Ipsen, H., Larsen, J. N., Joost van Neerven, R. J., Schou, C., Løwenstein, H. and Spangfort, M. D. 1996. X-ray and NMR structure of Bet v 1, the origin of birch pollen allergy. Nature Struct. Biol. 3: 1040-1045.PubMedGoogle Scholar
  20. Ganglberger, E., Grunberger, K., Sponer, B., Radauer, C., Breiteneder, H., Boltz-Nitulescu, G., Scheiner, O. and Jensen-Jarolim, E. 2000. Allergen mimotopes for 3-dimensional epitope search and induction of antibodies inhibiting human IgE. FASEB J. 14: 2177-2184.PubMedGoogle Scholar
  21. Gleave, A. P., Mitra, D. S., Markwick, N. P., Morris, B. A. M. and Beuning, L. L. 1998. Enhanced expression of the Bacillus thuringiensis cry9Aa2 gene in transgenic plants by nucleotide sequence modification confers resistance to potato tuber moth. Mol. Breed. 4: 459-472.Google Scholar
  22. Gonneau, M., Pagant, S., Brun, F. and Laloue, M. 2001. Photoaffinity labelling with the cytokinin agonist azido-CPPU of a 34 kDa peptide of the intracellular pathogenesisrelated protein family in the moss Physcomitrella patens. Plant Mol. Biol. 46: 539-548.PubMedGoogle Scholar
  23. Guilford, P., Prakash, S., Zhu, J. M., Rikkerink, E., Gardiner, S., Bassett, H. and Forster, R. (1997). Microsatellites in Malus × domestica (apple): abundance, polymorphism and cultivar identification. Theor. Appl. Genet. 94:249-254.Google Scholar
  24. Helsper, J. P. F. G., Gilissen, L. J. W. J., van Ree, R., America, A. H. P., Cordewener, J. H. G. and Bosch, D. 2002. Quadrupole time-of-flight mass spectrometry: a method to study the actual expression of allergen isoforms identified by PCR cloning. J. Allergy Clin. Immunol. 110: 131-138.PubMedGoogle Scholar
  25. Hoffmann-Sommergruber, K. 2002. Pathogenesis-related (PR)-proteins identified as allergens. Biochem. Soc. Trans. 30: 930-935.PubMedGoogle Scholar
  26. Hoffmann-Sommergruber, K., Vanek-Krebitz, M., Radauer, C., Wen, J., Ferreira, F., Scheiner, O. and Breiteneder, H. 1997. Genomic characterization of members of the Bet v 1 family: genes coding for allergens and pathogenesis-related proteins share intron positions. Gene 197: 91-100.PubMedGoogle Scholar
  27. Hsieh, L.-S., Moos, M., Jr and Lin, Y. 1995. Characterisation of apple 18 and 31 kd allergens by microsequencing and evaluation of their content during storage and ripening. J. Allergy Clin. Immunol. 96: 960-970.PubMedGoogle Scholar
  28. Knox, B. and Suphioglu, C. 1996. Environmental and molecular biology of pollen allergens. Trends Plant Sci. 1: 156-164.Google Scholar
  29. Laffer, S., Hamdi, S., Lupinek, C., Sperr, W. R., Valent, P., Verdino, P., Keller, W., Grote, M., Hoffmann-Sommergruber, K., Scheiner, O., Kraft, D., Rideau, M. and Valenta, R. 2003. Molecular characterization of recombinant T1, a nonallergenic periwinkle (Catharanthus roseus) protein, with sequence similarity to the Bet v 1 plant allergen family. Biochem. J. 373: 261-269.PubMedGoogle Scholar
  30. Lahti, A., Bjorksten, F. and Hannuksela, M. 1980. Allergy to birch pollen and apple, and cross-reactivity of the allergens studied with the RAST. Allergy 35: 297-300.PubMedGoogle Scholar
  31. Langenkämper, G., McHale, R., Gardner, R. C. and MacRae, E. 1998. Sucrose-phosphate synthase steady-state mRNA increases in ripening kiwifruit. Plant Mol. Biol. 36: 857-869.PubMedGoogle Scholar
  32. Liu, J.-J. and Ekramoddoullah, A. K. M. 2003. Root-specific expression of a western white pine PR10 gene is mediated by different promoter regions in transgenic tobacco. Plant Mol. Biol. 52: 103-120.PubMedGoogle Scholar
  33. Maliepaard, C., Alston, F. H., van Arkel, G., Brown. L. M., Chevreau. E., Duneman, F., Evans, K. M., Gardiner, S., Guilford, P., van Heusden, A. W., Janse, J., Laurens, F., Lynn, J. R., Manganaris, A. G., den Nijs, A. P. M., Periam, N., Rikkerink, E., Roche, P., Ryder, C., Sansavini, S., Schmidt, H., Tartarini, S., Verhaegh, J. J., Vrielink-van Ginkel, M. and King, G. J. (1998). Aligning male and female linkage maps of apple (Malus pumila Mill. ) using multi-allelic markers. Theor. Appl. Genet. 97: 60-73.Google Scholar
  34. Markovic-Housley, Z., Degano, M., Lamba, D., von Roepenack-Lahaye, E., Clemens, S., Susani, M., Ferreira, F., Scheiner, O. and Breiteneder, H. 2003. Crystal structure of a hypoallergenic isoform of the major birch pollen allergen Bet v 1 and its likely biological function as a plant steroid carrier. J. Mol. Biol. 325: 123-133.PubMedGoogle Scholar
  35. Matton, D. P. and Brisson, N. 1989. Cloning, expression, and sequence conservation of pathogenesis-related gene transcripts of potato. Mol. Plant Microbe Interact. 2: 325-331.PubMedGoogle Scholar
  36. Mogensen, J. E., Wimmer, R., Larsen, J. N., Spangfort, M. D. and Otzen, D. E. 2002. The major birch allergen, Bet v 1, shows affinity for a broad spectrum of physiological ligands. J. Biol. Chem. 277: 23684-23692.PubMedGoogle Scholar
  37. Neudecker, P., Schweimer, K., Nerkamp, J., Scheurer, S., Vieths, S., Stic, H. and Roschy, P. 2001. Allergic crossreactivity made visible: solution structure of the major cherry allergen Pruav 1. J. Biol. Chem. 276: 22756-22763.PubMedGoogle Scholar
  38. Neudecker, P., Lehmann, K., Nerkamp, J., Haase, T., Wangorsch, A., Fotisch, K., Hoffmann, S., Rosch, P., Vieths, S. and Scheurer, S. 2003. Mutational epitope analysis of Pru av 1 and Api g 1, the major allergens of cherry (Prunus avium) and celery (Apium graveolens): correlating IgE reactivity with three-dimensional structure. Biochem. J. 376: 97-107.PubMedGoogle Scholar
  39. Neuhoff, V., Norbert, A., Taube, D. and Ehrhardt, W. 1988. Improved staining of proteins following separation in polyacrylamide gels including isoelectric focusing gels with clear background at nanogram sensitivity using Coomassie Brilliant Blue G-250 and R-250. Electrophoresis 9: 255-262.PubMedGoogle Scholar
  40. Noiton, D. A. M. and Alspach, P. A. 1996. Founding clones, inbreeding, coancestry, and status number of modern apple cultivars. J. Am. Soc. Hort. Sci. 121: 773-782.Google Scholar
  41. Oraguzie, N. C., Gardiner, S. E., Basset, H. C. M., Stefanati, M., Ball, R. D., Bus, V. G. M. and White, A. G. 2001. Genetic diversity and relationships in Malus sp. germplasm collections as determined by randomly amplified polymorphic DNA. J. Am. Soc. Hort. Sci. 126: 318-328.Google Scholar
  42. Poupard, P., Strullu, D.-G. and Simoneau, P. 1998. Two members of the Bet v 1 gene family encoding birch pathogenesis-related proteins display different patterns of root expression and wound-inducibility. Aust. J. Plant Physiol. 25: 459-464.Google Scholar
  43. Pühringer, H., Moll, D., Hoffmann-Sommergruber, K., Watillon, B., Katinger, H. and Laimer da Camara Machado, M. 2000. The promoter of an apple Ypr10 gene, encoding the major allergen Mal d 1, is stress-and pathogen-inducible. Plant Sci. 152: 35-50.Google Scholar
  44. Pühringer H. M., Zinoecker, I., Marzban, G., Katinger, H. and Laimer, M. 2003. MdAP, a novel protein in apple, is associated with the major allergen Mal d 1. Gene 321: 173-183.PubMedGoogle Scholar
  45. Saraste, M., Sibbald, P. R. and Wittinghofer, A. 1990. The Ploop-a common motif in ATP-and GTP-binding proteins. TIBS 15: 430-434.PubMedGoogle Scholar
  46. Scheurer, S., Son, D. Y., Boehm, M., Karamloo, F., Franke, S., Hoffmann, A., Haustein, D. and Vieths, S. 1999. Crossreactivity and epitope analysis of Pru av 1, the major cherry allergen. Mol. Immun. 26: 155-167.Google Scholar
  47. Schöning, B., Ziegler, W. H., Vieths, S. and Baltes, W. 1996. Apple allergy: the cDNA sequence of the major allergen of apple, determined by performing PCR with a primer based on the N-terminal amino acid sequence, is highly homologous to the sequence of the major birch pollen allergen. J. Sci. Food Agric. 71: 475-482.Google Scholar
  48. Simon, C. J. and Weeden, N. F. 1992. Molecular analysis and cloning of Malus ribosomal DNA. J. Am. Soc. Hort. Sci. 117:164-168.Google Scholar
  49. Son, D. Y., Scheurer, S., Hoffmann, A., Haustein, D. and Vieths, S. 1999. Pollen-related food allergy: cloning and immunological analysis of isoforms and mutants of Mal d 1, the major apple allergen, and Bet v 1, the major birch pollen allergen. Eur. J. Nutr. 38: 201-215.PubMedGoogle Scholar
  50. Sonnewald, U., Quick, W. P., MacRae, E., Krause, K. P. and Stitt, M. 1993. Purification, cloning and expression of spinach leaf sucrose-phosphate synthase in Escherichia coli. Planta 189: 174-181.PubMedGoogle Scholar
  51. Spangfort, M. D., Ipsen, H., Sparholt, S. H., Aasmul-Olsen, S., Osmark, P., Poulsen, F. M., Larsen, M., Mortz, E., Roepstorff, P. and Larsen, J. N. 1996. Characterisation of recombinant isoforms of birch pollen allergen Bet v 1. Adv. Exp. Med. Biol. 409: 251-254.PubMedGoogle Scholar
  52. Swoboda, I., Scheiner, O., Kraft, D., Breitenbach, M., Heberle-Bors, E. and Vicente, O. 1994. A birch gene family encoding pollen allergens and pathogenesis-related proteins. Biochim. Biophys. Acta 1219: 457-464.PubMedGoogle Scholar
  53. Swoboda, I., Scheiner, O., Heberle-Bors, E. and Vicente, O. 1995a. cDNA cloning and characterization of three genes in the Bet v 1 gene family that encode pathogenesis-related proteins. Plant, Cell Environ. 18: 865-874.Google Scholar
  54. Swoboda, I., Jilek, A., Ferreira, F., Engel, E., Hoffmann-Sommergruber, K., Scheiner, O., Kraft, D., Breiteneder, H., Pittenauer, E., Schmid, E., Vicente, O., Heberle-Bors, E., Ahorn, H. and Breitenbach, M. 1995b. Isoforms of Bet v 1, the major birch pollen allergen, analyzed by liquid chromatography, mass spectrometry, and cDNA cloning. J. Biol. Chem. 270: 2607-2613.PubMedGoogle Scholar
  55. Swoboda, I., Hoffman-Sommergruber, K., O'Riordain, G., Scheiner, O., Heberle-Bors, E. and Vicente, O. 1996. Bet v 1 proteins, the major birch pollen allergens and members of a family of conserved pathogenesis-related proteins, show ribonuclease activity in vitro. Physiol. Plant 96: 433-438.Google Scholar
  56. Towbin, H., Staehelin, T. and Gordon, J. 1979. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc. Natl Acad. Sci. USA 76: 4350-4354.PubMedGoogle Scholar
  57. Utriainen, M., Kokko, H., Auriola, S., Sarrazin, O. and Kärenlamp, S. 1998. PR-10 protein is induced by copper stress in roots and leaves of a Cu/Zn tolerant clone of birch, Betula pendula. Plant Cell Environ. 21: 821-828.Google Scholar
  58. van Ree, R. and Aalberse, R. C. 1993. Pollen-vegetable food crossreactivity: serological and clinical relevance of crossreactive IgE. J. Clin. Immunoassay 16: 124-130.Google Scholar
  59. Vanek-Krebitz, M., Hoffmann-Sommergruber, K., Laimer da Camara Machado, M., Susani, M., Ebner, C., Kraft, D., Scheiner O. and Breiteneder, H. 1995. Cloning and sequencing of Mal d 1, the major allergen from apple (Malus domestica), and its immunological relationship to Bet v 1, the major birch pollen allergen. Biochem. Biophys. Research Commun. 214: 538-551.Google Scholar
  60. Vieths, S., Schöning, B. and Jankiewicz, A. 1993. Occurrence of IgE binding allergens during ripening of apple fruits. Food Agric. Immunol. 5: 93-105.Google Scholar
  61. Vieths, S., Schöning, B. and Peterson, A. 1994a. Characterization of the 18-kDa apple allergen by two-dimensional immunoblotting and microsequencing. Int. Arch. Allergy Immunol. 104: 399-404.PubMedGoogle Scholar
  62. Vieths, S., Jankiewicz, A., Schöning, B. and Aulepp, H. 1994b. Apple allergy: the IgE-binding potency of apple strains is related to the occurrence of the 18-kDa allergen. Allergy 49: 262-271.PubMedGoogle Scholar
  63. Vieths, S., Scheurer, S. and Ballmer-Weber, B. 2002. Current understanding of cross-reactivity of food allergens and pollen. Ann. NY. Acad. Sci. 964: 47-68.PubMedGoogle Scholar
  64. Walter, M. H., Liu, J.-W., Wünn, J. and Hess, D. 1996. Bean ribonuclease-like pathogenesis-related protein genes (Ypr10) display complex patterns of developmental, dark-induced and exogenous-stimulus-dependent expression. Eur. J. Biochem. 239: 281-293.PubMedGoogle Scholar
  65. Yu, X., Ekramoddoullah, A. K. M. and Misra, S. 2000. Characterization of Pin m III cDNA in western pine. Tree Physiol. 20: 663-671.PubMedGoogle Scholar
  66. Ziadi S., Poupard P., Brisset M-N., Paulin J. and Simoneau P. 2001. Characterisation in apple leaves of two subclasses of PR-10 transcripts inducible by acibenzolar-S-methyl, a functional analogue of salicylic acid. Physiol. Mol. Plant Path. 59: 33-43.Google Scholar

Copyright information

© Kluwer Academic Publishers 2004

Authors and Affiliations

  • Lesley Beuning
    • 1
  • Judith Bowen
    • 1
  • Helena Persson
    • 2
  • Diane Barraclough
    • 1
  • Sean Bulley
    • 1
  • Elspeth MacRae
    • 1
  1. 1.The Horticulture and Food Research Institute of New Zealand Ltd.Mt Albert Research CentrePrivate BagNew Zealand
  2. 2.SLU-Balsgård, Department of Crop ScienceSwedish University of Agricultural SciencesSweden

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