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Abstract
A detailed histo-anatomical and phytochemical study on Globularia alypum was carried out to investigate season-induced plant responses related to leaf morphology and content of secondary metabolites (SMs). Leaf tissue analysis by light and scanning electron microscopy showed neither morphological nor significant anatomical adaptions/variations to the coldest and the hottest seasons. Both summer and winter leaf tissue resulted amphistomatic with compact mesophyll and capitate glandular trichomes. Epidermal and mesophyll cells showed a significant accumulation of osmiophilic compounds, further characterized as phenolics by the histochemical screening. Beside the morpho-anatomical similarities in tissue arrangement, SMs production, within and among the different natural product classes, resulted seasonally dependent. In total, 24 SMs belonging to iridoids, flavonoids and phenylethanoid glycosides were tentatively assigned by UHPLC-HRMS analysis. Among iridoids, (E)-globularicisin showed to be the prevalent constituent, with content up to 70%. Total iridoid production was increased by 48.1% during summer. Similarly, flavonoid anabolism was more active during the same period, although with less remarkable seasonal variation reaching 25.6%. In contrast to the overall higher production of iridoids and flavonoids, a drastic fall (33.5%) of phenylethanoids was observed during summer.