BBAP Amplification Profiles of Golden Delicious and Granny Smith Apple Cultivars

Abstract

The BBAP analysis (Bet v 1 based amplified polymorphism) used in the study was aimed to detect natural variation among thirteen samples of Golden Delicious and Granny Smith apple cultivars. These cultivars were selected due to their global commercial importance and their documented association with high concentrations of the allergen Mal d 1. All of them were sourced from retail chains across eight European countries (Germany, Austria, Slovakia, Serbia, Croatia, Hungary, Italy, and Slovenia). Cross-reactivity between Bet v 1, a major allergen in birch pollen, and Mal d 1, its homolog in apples, often leads to allergic reactions in individuals sensitized to birch pollen. That is why determining the features of the genome of different cultivars of apples with the use of appropriate specific primers makes it possible to assess the possible allergenicity of products. Forward and reverse primers (R1, R2, R3, R4, Rd) were employed, and the resulting amplification products were analysed via electrophoresis on a 3% agarose gel. Most samples exhibited monomorphic patterns, suggesting similar isoform profiles, though some differences between cultivars were observed. A total of 73 fragments, ranging from 38 bp to 662 bp, were amplified. Profiles for primers FR3 and FR4 could not be successfully synthesized. For the primer pair FRd, fragment sizes ranged from 113 bp to 400 bp, with 26 fragments observed. Primer pair FR1 produced 20 fragments within a range of 181 bp to 662 bp, while primer pair FR2 resulted in 27 fragments ranging from 38 bp to 307 bp. All primer pairs exhibited the expected amplicon with a length of 388 bp. An average PIC value of 0.37 confirmed the high informativeness of the technique. The monomorphic profiles suggest that the environmental impact and varying storage conditions on the studied genes are minimal or negligible, as all samples exhibited similar profiles despite being sourced from diverse countries and retail stores. The identification of unique genetic profiles in certain regions or cultivars may provide insights into the allergenic potential of different apple cultivars, contributing to advancements in food safety, allergen management, and the development of hypoallergenic apple cultivars.