Characterisation of Mal d 1-related genes in Malus
- 1 May 2004
- journal article
- Published by Springer Nature in Plant Molecular Biology
- Vol. 55 (3) , 369-388
- https://doi.org/10.1007/s11103-004-0904-9
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.Keywords
This publication has 64 references indexed in Scilit:
- Gapped BLAST and PSI-BLAST: a new generation of protein database search programsNucleic Acids Research, 1997
- X-ray and NMR structure of Bet v 1, the origin of birch pollen allergyNature Structural & Molecular Biology, 1996
- Apple Allergy: The cDNA Sequence of the Major Allergen of Apple, Determined by Performing PCR with a Primer Based on theN-Terminal Amino Acid Sequence, is Highly Homologous to the Sequence of the Major Birch Pollen AllergenJournal of the Science of Food and Agriculture, 1996
- A stress‐, pathogenesis‐, and allergen‐related cDNA in apple fruit is also ripening‐relatedNew Zealand Journal of Crop and Horticultural Science, 1996
- 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 AllergenBiochemical and Biophysical Research Communications, 1995
- A simple and efficient method for isolating RNA from pine treesPlant Molecular Biology Reporter, 1993
- Occurrence of IgE binding allergens during ripening of apple fruitsFood and Agricultural Immunology, 1993
- Cloning, Expression, and Sequence Conservation of Pathogenesis-Related Gene Transcripts of PotatoMolecular Plant-Microbe Interactions®, 1989
- Improved staining of proteins in polyacrylamide gels including isoelectric focusing gels with clear background at nanogram sensitivity using Coomassie Brilliant Blue G‐250 and R‐250Electrophoresis, 1988
- Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.Proceedings of the National Academy of Sciences, 1979