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Background  Mal d 1 is a major apple allergen causing food allergic symptoms of the oral allergy syndrome (OAS) in birch-pollen sensitised patients. The  Mal d 1  gene family is known to have at least 7 intron-containing and 11 intronless members that have been mapped in clusters on three linkage groups. In this study, the allelic diversity of the seven intron-containing  Mal d 1  genes was assessed among a set of apple cultivars by sequencing or indirectly through pedigree genotyping. Protein variant constitutions were subsequently compared with  S kin  P rick  T est (SPT) responses to study the association of deduced protein variants with allergenicity in a set of 14 cultivars.    Results  From the seven intron-containing  Mal d 1  genes investigated,  Mal d 1.01  and  Mal d 1.02  were highly conserved, as nine out of ten cultivars coded for the same protein variant, while only one cultivar coded for a second variant.  Mal d 1.04 ,  Mal d 1.05  and  Mal d 1.06 A, B  and  C  were more variable, coding for three to six different protein variants. Comparison of  Mal d 1  allelic composition between the high-allergenic cultivar Golden Delicious and the low-allergenic cultivars Santana and Priscilla, which are linked in pedigree, showed an association between the protein variants coded by the  Mal d 1.04  and  -1.06A  genes (both located on linkage group 16) with allergenicity. This association was confirmed in 10 other cultivars. In addition,  Mal d 1.06A  allele dosage effects associated with the degree of allergenicity based on prick to prick testing. Conversely, no associations were observed for the protein variants coded by the  Mal d 1.01  (on linkage group 13), - 1.02 , - 1.06B, -1.06C  genes (all on linkage group 16), nor by the  Mal d 1.05  gene (on linkage group 6).    Conclusion  Protein variant compositions of Mal d 1.04 and -1.06A and, in case of  Mal d 1.06A , allele doses are associated with the differences in allergenicity among fourteen apple cultivars. This information indicates the involvement of qualitative as well as quantitative factors in allergenicity and warrants further research in the relative importance of quantitative and qualitative aspects of  Mal d 1  gene expression on allergenicity. Results from this study have implications for medical diagnostics, immunotherapy, clinical research and breeding schemes for new hypo-allergenic cultivars.

Assessment of allelic diversity in intron-containing Mal d 1 genes and their association to apple allergenicity