Investigating the simultaneous effect of chitosan and arbuscular mycorrhizal fungi on growth, phenolic compounds, PAL enzyme activity and lipid peroxidation in Salvia nemorosa L.

Considering the importance of Salvia nemorosa L. in the pharmaceutical and food industries, and also beneficial approaches of arbuscular mycorrhizal fungi (AMF) symbiosis and the use of bioelicitors such as chitosan to improve secondary metabolites, the aim of this study was to evaluate the performance of chitosan on the symbiosis of AMF and the effect of both on the biochemical and phytochemical performance of this plant and finally introduced the best treatment. Two factors were considered for the factorial experiment: AMF with four levels (non-inoculated plants, Funneliformis mosseae, Rhizophagus intraradices and the combination of both), and chitosan with six levels (0, 50, 100, 200, 400 mg L-1 and 1% acetic acid). Four months after treatments, the aerial part and root length, the levels of lipid peroxidation, H2O2, phenylalanine ammonia lyase (PAL) activity, total phenol and flavonoid contents and the main secondary metabolites (rosmarinic acid and quercetin) in the leaves and roots were determined. The flowering stage was observed in R. intraradices treatments and the highest percentage of colonization (78.87%) was observed in the treatment of F. mosseae × 400 mg L-1 chitosan. Furthermore, simultaneous application of chitosan and AMF were more effective than their separate application to induce phenolic compounds accumulation, PAL activity and reduce oxidative compounds. The cluster and principal component analysis based on the measured variables indicated that the treatments could be classified into three clusters. It seems that different treatments in different tissues have different effects. However, in an overview, it can be concluded that 400 mg L-1 chitosan and F. mosseae × R. intraradices showed better results in single and simultaneous applications. The results of this research can be considered in the optimization of this medicinal plant under normal conditions and experiments related to abiotic stresses in the future.

Phenolics and terpenoids change in response to yeast extract and chitosan elicitation in Zataria multiflora cell suspension culture.

Zataria multiflora is an important medicinal plant with antioxidant and anticancer properties attributed to its phytochemicals. To develop a method for bulk production of valuable phytochemicals, cell suspension culture of Z. multiflora were grown in liquid B5 medium and then treated in their log growth phase with chitosan (0, 10, 20, and 40 mg L-1) and yeast extract (0, 400, 800, and 1200 mg L-1) for 3 days. The levels of hydrogen peroxide (H2O2), nitric oxide (NO), malondialdehyde (MDA), and the main terpenoids and phenylpropanoids in the cell extracts were determined by HPLC and spectrophotometric techniques. The H2O2 and MDA levels significantly increased in the cells treated with both yeast extract and chitosan, while the NO level increased in those exposed to yeast extract. At their highest concentrations, both elicitors significantly increased PAL and TAL activities, as well as phenolic acids and flavonoids contents. Chitosan only induced the production of caffeic acid (22 µg g-1 DW), benzoic acid (2 µg g-1 DW), 4-hydroxy benzoic acid (6 µg g-1 DW), epicatechin (63 µg g-1 DW), and apigenin (5 µg g-1 DW) in the cells, while yeast extract increased the contents of phenylpropanoids gallic acid (50 µg g-1 DW), vanillin (35 µg g-1 DW), salicylic acid (24 µg g-1 DW), catechin (130 µg g-1 DW) and terpenoids carvacrol (7 µg g-1 DW) and thymol (24 µg g-1 DW). In conclusion, changes in the production of phenolics and terpenoids are a defensive mechanism in Z. multiflora cells treated by yeast extract and chitosan. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-022-03235-x.