Abstract
Sorbus domestica L. leaves are a phenylpropanoid-rich herbal medicine with antioxidant and anti-inflammatory activity. Here, to optimise harvesting time of the leaves for medicinal application, seasonal variability in the expression of genes encoding enzymes involved in phenylpropanoid biosynthesis (qRT-PCR, quantitative real time polymerase chain reaction) and its relationship with changes in the phytochemical profile (UHPLC-PDA-ESI-MS3, HPLC-PDA-fingerprint, spectrophotometric methods) and antioxidant properties (2,2-diphenyl-1-picrylhydrazyl scavenging and ferric reducing antioxidant power) was evaluated during foliar development. Significant quantitative differences in polyphenolic composition were revealed between spring leaves (predominance of phenylpropanoid acids and flavonoids) and those harvested during the rest of the growing season (predominance of proanthocyanidins, major active components of S. domestica), while their qualitative composition was stable. A set of 40 compounds were detected in the samples by LC–MS/MS including 38 phenylpropanoids. The total phenylpropanoid level (HPLC) in dried leaves dropped from 25.7 mg/g in May to 20.7 mg/g in October but proanthocyanidins and the antioxidant capacity increased simultaneously up to 12.7 mg/g and 49.7 mmol Trolox equivalents/100 g, respectively. Based on the correlation between gene expression and total phenolics and proanthocyanidins (r > 0.825, p < 0.05), phenylalanine ammonia lyase (PAL) and 4-coumaroyl:CoA ligase (4CL) appeared to be core regulators of the phenolic profile among analysed genes (PAL; 4CL; C4H, cinnamate 4-hydroxylase; CHS, chalcone synthase; F3H, flavanone 3-hydroxylase; FLS, flavonols synthase). Due to the synergy of individual components, variability in activity parameters was relatively moderate. Considering the phenylpropanoid levels and antioxidant capacity, the late summer and autumn months (August-October) might be the most advantageous time for harvesting leaf material of optimal quality.
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