Introduction – Very rarely fruit pulp has been used in in vitro culture to produce secondary metabolites useful in promoting
health.
Objectives – The aims of this work were the study of the best conditions to obtain the callus cultures from the pulp of two
varieties of apples, Golden Delicious (GD) and “Mela Rosa Marchigiana” (MRM), and the quali-quantitative analysis of secondary
metabolites produced by the two in vitro callus cultures.
Methodology – Callus was induced on both Murashige and Skoog and Gamborg B5 media containing various combinations of
supplements. To achieve the maximum recovery of secondary metabolites produced, preliminary extraction tests were carried
out on GD apple culture using two different organic solvents (MeOH and EtOAc). The quali-quantitative analysis of the methanolic
extract of both cultures was carried out by ESI-MSn and GC–MS techniques.
Results – The GC–MS analysis revealed the presence of triterpenic acids, in particular, oleanolic, ursolic, maslinic, pomolic,
tormentic, corosolic and annurcoic acid along with a phytosterol, β-sitosterol. In addition, GD callus culture produced phloridzin,
absent in the MRM culture. In this last culture, however, the total amount of secondary metabolites was markedly higher. The
in vivo production of these bioactive compounds were also quantified in the GD and MRM apple pulps.
Conclusion – Apple pulps produced higher amounts of triterpenic acids in vitro than in vivo. The presentwork can be considered a
method to amplify the production of important secondary metabolites which exert beneficial effects on human health.
