Abstract
Non-specific lipid transfer proteins (nsLTPs) of Rosaceae fruits, such as peach, apricot, cherry, plum and apple, represent major allergens for Mediterranean atopic populations. As a first step in elucidating the genetics of nsLTPs, we directed the research reported here towards identifying the number and location of nsLTP (Mal d 3) genes in the apple genome and determining their allelic diversity. PCR cloning was initially performed on two cultivars, Prima and Fiesta, parents of a core apple mapping progeny in Europe, based on two Mal d 3 sequences (AF221502 and AJ277164) in the GenBank. This resulted in the identification of two distinct sequences (representing two genes) encoding the mature nsLTP proteins. One is identical to accession AF221502 and has been named Mal d 3.01, and the other is new and has been named Mal d 3.02. Subsequent genome walking in the upstream direction and DNA polymorphism analysis revealed that these two genes are intronless and that they could be mapped on two homoeologous segments of linkage groups 12 and 4, respectively. Further cloning and sequencing of the coding and upstream region of both Mal d 3 genes in eight cultivars was performed to identify allelic variation. Assessment of the deduced nsLTP amino acid sequences gave a total of two variants at the protein level for Mal d 3.01 and three for Mal d 3.02. The consequences of our results for allergen nomenclature and the breeding of low allergenic apple cultivars are discussed.
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Acknowledgements
This research was funded in part by the EU (EU-SAFE, QLK1-CT-2000-01394). We took the advantage of using the updated molecular maker linkage map of Prima and Fiesta from the European DARE project (FAIR5-CT97-3898). The authors thank Professors Evert Jacobsen and Richard Visser for their critical reading of this manuscript and their stimulating interest. The first author is grateful to ICSC-World Laboratory, Lausanne for providing a one-year scholarship in 2001.
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Gao, Z.S., van de Weg, W.E., Schaart, J.G. et al. Linkage map positions and allelic diversity of two Mal d 3 (non-specific lipid transfer protein) genes in the cultivated apple (Malus domestica). Theor Appl Genet 110, 479–491 (2005). https://doi.org/10.1007/s00122-004-1856-9
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DOI: https://doi.org/10.1007/s00122-004-1856-9