The use of silicon and mycorrhizal fungi to mitigate changes in licorice leaf micromorphology, chlorophyll fluorescence, and rutin content under water-deficit conditions

Abstract

In this study, the effects of water-deficit conditions, silicon (Si) fertilizer (300 ppm), and arbuscular mycorrhizal (AM) inoculation by Claroiedoglomus etunicatum were evaluated on several features of licorice (Glycyrrhiza glabra L.). The measurable features were photosynthetic parameters, rutin content in aerial parts, and leaf micromorphology. Drought was administered at five levels determined by the percentage of field capacity (FC), i.e. 100, 80, 60, 40, and 20% of FC. Leaf extracts were utilized for measuring rutin content (via HPLC), and photosynthetic pigments; measurement of stomatal density, and trichome analysis were performed by scanning electron microscopy (SEM). Under severe drought stress, leaf area decreased by 50.84%, compared to well-irrigated plants. A significant decrease in leaf numbers (32.52%) was observed because of deficit irrigation. AM and Si improved chlorophyll fluorescence, which corresponded to the maximum efficiency of photosystem II. Rutin content decreased significantly under deficit irrigation. Also, the integration of AM and Si treatments positively affected rutin quantity under various irrigation regimes. Under moderate stress (60% FC), using AM and/or Si treatments reduced the stomatal length by 61.22 and 52.98%, respectively. Interestingly, a significant reduction in stomatal density towards control was observed as a result of the integrated treatments of Si and AM (58.28% at W20 and 59.82% at W100), which helped plants reduce water loss when facing drought stress. Principal component analysis (PCA) showed that photosynthetic pigments, chlorophyll fluorescence, and rutin changed quantitatively under moderate drought stress, while more variations were observed in leaf epidermal micromorphology under severe drought stress. These findings revealed that Si and AM, by exogenous application, synergistically mitigated the effects of drought stress on licorice.