Modulation of anthraquinones and phloroglucinols biosynthesis in Hypericum spp. by cryogenic treatment

Beside the high post-cryogenic recovery rate, a reinstated scale of secondary metabolites in recovered plant tissues represents another inevitable aspect of an effective cryopreservation protocol for medicinal plants. The current study was aimed at evaluation of the elicitation potential of cryogenic treatment on secondary metabolism of some Hypericum species. In agreement with our assumption, the cold stimuli applied during the pre-cryogenic phase increased the tolerance to low temperatures (-196°C) in H. perforatum, H. rumeliacum and H. tetrapterum reaching a maximum of 46% recovery rate in St. John's wort plants. The effect of cryogenic treatment-associated stressors on the spectrum of the profiling secondary metabolites, naphthodianthrones and phloroglucinols, was ambiguous. The content of hypericins in both pre-cultured H. tetrapterum donor plants and H. perforatum shoots regenerated from cryopreserved meristems increased more than 3-times. The highest 38-fold enhancement of phloroglucinols was observed in H. rumeliacum shoots recovered after cryostorage. Our findings indicate that modulated biosynthesis of secondary metabolites represented by naphtodianthrones and phloroglucinols can be considered as a part of overall plant adaptations to stress conditions associated with liquid nitrogen (LN) treatment.