Short-Term Liquid Nitrogen Storage of Pyrostegia venusta Embryos: Effects on Germination, Phenotypic and Biochemical Characteristics, and In Vitro Secondary Metabolite Production

Abstract

In this study, short-term liquid nitrogen (LN) storage was used as a strategy to conserve Pyrostegia venusta embryos, and its effects on in vitro germination, phenotypic and biochemical characteristics, and in vitro secondary metabolite production were assessed. Embryos stored in LN for 1 and 7 days presented a higher germination rate and germination speed index compared to those of the control (non-cryostored embryos). Short-term LN storage also favored the phenotypic characteristics of seedlings. LN storage significantly affected the proteins (PTN), soluble sugar (SS) and reducing sugar (RS) contents, oxidative metabolism, and phenylalanine ammonia-lyase (PAL) activity, as well as the total phenolic compound, flavonoid, phytosterol, and alkaloid levels in seedlings regenerated from embryos cryostored for 7 days. Benzoic acid derivatives and flavonoids were observed in regenerated non-acclimatized seedlings. LN storage did not affect the survival rate or phenotypic characteristics of seedlings during acclimatization. Acclimatization promoted significant changes in PTN, SS and RS contents, oxidative metabolism, and PAL activity in seedlings from embryos cryostored for 7 days. Roots from acclimatized seedlings exhibited the highest phenolic, phytosterol, and total alkaloid levels. Differences in the chromatographic profiles of the acclimatized seedlings compared with the non-acclimatized seedlings were observed. LN storage can be an effective means of ex situ conservation of P. venusta genetic resources.