Beneficial effects of endophytic fungi inoculation on tanshinones and phenolic compounds of Salvia abrotanoides.

Objectives: Salvia abrotanoides is considered as a new source of tanshinone-producing plants in Iran. Symbiosis of endophytic fungi with their host plants is an effective tool to promote the growth and secondary metabolism of medicinal herbs. Therefore, using endophytic fungi as a biotic elicitor is a proper solution to increase the yield of plant products. Materials and Methods: In this study, some endophytic fungi were first isolated from the root of S. abrotanoides, then two of them (Penicillium canescens and Talaromyces sp.) were co-cultivated with the sterile seedling of S. abrotanoides in pot culture. After proving the colonization of these fungi in the root tissues by microscopic studies, their effects on the production of critical medicinal compounds such as tanshinones and phenolic acids were investigated in the vegetation stage (120 days). Results: Our results showed that the content of cryptotanshinone (Cry) and tanshinone IIA (T-IIA) in plants inoculated with P. canescens increased by 77.00% and 19.64%, respectively, compared with non-inoculated plants (control). The contents of mentioned compounds in plants inoculated with Talaromyces sp. increased by 50.00% and 23.00%, respectively. In this case, in plants inoculated with P. canescens, it was found that the level of caffeic acid, rosmarinic acid, and its PAL enzyme activity increased by 64.00%, 69.00%, and 50.00%, respectively, compared with the control. Conclusion: Endophytic fungi have specific modes of action and the ability to provide multiple benefits. Each of the two strains is a highly considerable microbial resource for the growth and accumulation of active compounds of S. abrotanoides.

The variability of phenolic constituents and antioxidant properties among wild populations of Ziziphora clinopodioides Lam.

In this study, phenolic derivatives and antioxidant activities of fourteen Ziziphora clinopodioides populations, as well as LC-MS/MS analysis of three specific flavonoids were evaluated. Generally, high contents of phenolic derivatives were found in shoot extracts compared to roots. LC-MS/MS, a powerful analytical technique, was employed for the identification and quantify the individual flavonoids in Z. clinopodioides populations' extracts, in a quantity order of quercetin > rutin > apigenin. Scavenging activity by DPPH and FRAP was performed, and accordingly, in the shoot, the highest values for the DDPH were 4.61 ± 0.4 and 7.59 ± 0.26 µg ml- 1 in populations 1 and 13, respectively, and for the FRAP were 328.61 ± 5.54 and 292.84 ± 2.85 mg g DW- 1, in populations 6 and 1 respectively. Multivariate analysis results of the principal component analysis indicated the amount of polyphenols to be useful indicators in differentiating the geographical localities which explain 92.7% of the total variance. According to the results of hierarchical cluster analysis, the studied populations could be separated into two groups in that the contents of phenolic derivatives and antioxidant activities of different plant parts. Both shoot and root samples were well discriminated with the orthogonal partial least squares discriminant analysis (R2X: 0.861; Q2: 0.47) model. The validity of the model was confirmed by using receiver operating characteristic curve analysis and permutation tests. Such data make an important addition to our current knowledge of Ziziphora chemistry and are decisive in the identification of germplasms with a homogeneous phytochemical profile, high chemical content and bioactivity. The present results could also be helpful for the potential application of Z. clinopodioides in different kinds of industries as natural antioxidants. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-023-01283-y.

Plant elicitation and TiO2 nanoparticles application as an effective strategy for improving the growth, biochemical properties, and essential oil of peppermint.

Mentha piperita L., which is an abundant source of essential oils (EO) and phenolic acids, is well known for its medicinal significance. The present research aimed to evaluate the impact of various concentrations of methyl jasmonate (MeJA; 0, 0.1, and 0.5 mM), titanium dioxide nanoparticles (TiO2 NPs; 0 and 150 mg L-1), and salicylic acid (SA; 0, 0.1, and 1 mM) on growth, EOs, and phenolic compounds of M. piperita L. The results demonstrated that the simultaneous application of SA (0.1 mM) and TiO2 NPs (150 mg L-1) enhanced shoot dry weight, the shoot length, and membrane stability index of peppermint by 56.17, 19.52, and 36%, respectively, compared to control. Moreover, phenolic content (76%), caffeic acid content (78%), rosmarinic acid content (87%), 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging ability (78%), and catalase (155%), ascorbate peroxidase activities (95%) were further improved by simultaneously applying MeJA (0.1 mM) and TiO2 NPs (150 mg L-1) compared to control. The highest menthol production (44.51%) was obtained with exogenous application of MeJA (0.1 mM) with 150 mg L-1 TiO2 NPs. The findings of the current study presented an ideal combination of TiO2 NPs with plant growth regulators for promoting antioxidant activities and increasing major components of EO in peppermint plants.

ICP-OES assessment of trace and toxic elements in Ziziphora clinopodioides Lam. from Iran by chemometric approaches.

The aim of the study was to evaluate the concentrations of some essential and toxic elements (including Ca, K, Mg, Na, Fe, Cu, Mn, Zn, Co, Mo, Pb, Ni, Cr, and Cd) in Ziziphora clinopodioides Lam. (endemic Iranian herb) from 14 different regions by ICP-OES (inductively coupled plasma-optical emission spectrometry) method followed by multivariate statistical analyses. The analytical performances were assessed as the limit of detection (LOD), limit of quantification (LOQ), precision, and accuracy. Multivariate analysis (CA, PCA and HCA) showed the elemental distribution in the roots were higher than the shoots and significant element was Ca between regions. Three principal components (PCs) explained 77.94% of the total variance. They were as follows: PC1 with Cu, Zn, K, Cr, Ni, and Mn; PC2 with Na, Ca, Pb, and Fe; and PC3 with Mg. Hierarchical cluster analysis indicated four groups of Z. clinopodioides samples from the 14 regions based on their trace and toxic element levels. These chemometric approaches with multivariate analysis enable researchers to understand and quantify the relationship between the variables in a data set, and the analysis considers more than one factor. The concentrations of Cu, Na, Mn, Zn, and Pb in most Z. clinopodioides samples were below the WHO (world health organization) limit for herbal medicines (10, 51,340, 200, 50, and 10 µg g-1 respectively), while Fe and Ca levels were higher than allowed (15 and 614 µg g-1 respectively). However, the WHO limit for Mg, K, Co, and Mo in medicinal plants has not yet been determined. The results of this study confirmed that different parts of Z. clinopodioides can be used as an important source for human nutrition due to its essential mineral elements.

Variation in phenolic compounds, α-linolenic acid and linoleic acid contents and antioxidant activity of purslane (Portulaca oleracea L.) during phenological growth stages.

Purslane (Portulaca oleracea L.) contains a variety of natural products with different biological properties. The objective of this research was to estimate changes in total phenolics, flavonoids, and fatty acids (α-linolenic acid and linoleic acid) contents as well as antioxidant activity of P. oleracea at different growth stages. Seeds were germinated in soil-filled plastic pots at greenhouse. Leaves and stems were collected at the vegetative and flowering stages. Total phenol and flavonoid contents of the samples were determined by Folin-Ciocalteau and aluminum chloride methods, respectively. The contents of α-linolenic and linoleic acids were determined using gas chromatography analysis after transesterification of fatty acids. Furthermore, Ferric reducing antioxidant power and 1,1-diphenyl-2-picrylhydrazyl assays were used to determine the antioxidant activities. The highest contents of total phenols (698.6 mg GAE 100 g-1 DW), flavonoids (46.9 mg QE 100 g-1 DW), α-linolenic acid (2.7 mg g-1 DW) and linoleic acid (0.8 mg g-1 DW) were determined in the leaves at flowering stage. Free radical scavenging capacity was significantly affected (P ≤ 0.05) by age; and the leaves of purslane had more antioxidant potential compared to stems. A positive correlation was observed between the antioxidant activities and total phenols content. Overall, purslane leaves at flowering stage can be regarded as a valuable source of fatty acids (especially α-linolenic acid) and antioxidants in human diet.

Optimization of Callus Induction with Enhancing Production of Phenolic Compounds Production and Antioxidants Activity in Callus Cultures of Nepeta binaloudensis Jamzad (Lamiaceae).

BACKGROUND: World Health Organization (WHO) reported that more than 80% of people in the world use herbal traditional medicines nowadays. Many endemic medicinal plants, especially Nepeta species, are facing to extinction as a result of high harvesting, limited distribution, and habitat destruction.Tissue culture is a successful method for plant secondary metabolites production. Nepeta binaloudensis is a medicinal plant belonging to family Lamiaceae. OBJECTIVE: Our study was focused on devising an optimum procedure for callus induction and phenolic compounds production in N. binaloudensis. First, we are focused on finding suitable explants and media for callus induction. Then, subsequent experiments were conducted to find an optimal concentration of plant growth regulators (PGRs) and reduced- glutathione for maximum biomass production, and phenolic compounds production in calli. MATERIAL AND METHOD: In this study, the usage of whole plant grown in Hoagland nutrient solution, were used as a source of explants. Also, different media including, ½ MS, MS, and B5 and different combination of PGRs (NAA and BAP) were used for optimization of calli induction. RESULTS: Based on the results of the first experiment, leaf-originated explants, and macro half strength MS (½ MS) medium were used for the next experiments. The highest FW (Fresh Weight) and DW (Dry Weight) of calli were observed in ½ MS medium, supplemented with 2 µM/L reduced-glutathione, 2 mg.L-1 BAP, and 2 mg.L-1 NAA. The maximum amount of total phenolic, flavonoid, tannin contents and free-radical scavenger were observed in calli which were grown in ½ MS medium supplemented with 2 µM/L reduced-glutathione, 2 mg.L-1 BAP, and 2 mg.L-1 NAA. CONCLUSION: Our study finds the optimum condition for calli induction and phenolic compounds production in N. binaloudensis.

Endophytic Fungi of Native Salvia abrotanoides Plants Reveal High Taxonomic Diversity and Unique Profiles of Secondary Metabolites.

Endophytic fungi are often embedded in their host's metabolic networks, which can result in alterations of metabolite production and higher amounts of active compounds in medicinal plants. This study reports the occurrence, diversity, and secondary metabolite profiles of endophytic fungi isolated from Salvia abrotanoides plants obtained from three geographically distinct sites in Iran. A total of 56 endophytic fungi were isolated from roots and leaves of S. abrotanoides; site-specificity and root-dominated colonization was found to be a general characteristic of the endophytes. Based on molecular identification, the endophytic fungi were classified into 15 genera. Mycelial extracts of these isolates were subjected to high-resolution mass spectrometry analyses and revealed a broad spectrum of secondary metabolites. Our results demonstrated that Penicillium canescens, P. murcianum, Paraphoma radicina, and Coniolariella hispanica are producers of cryptotanshinone, which is a main bioactive compound of S. abrotanoides. Moreover, it was shown that it can be produced independent of the host plant. The effect of exogenous gibberellin on S. abrotanoides and endophytic fungi was shown to have a positive effect on increasing the cryptotanshinone production in the plant as well as in endophytic fungi cultivated under axenic conditions. Our findings provide further evidence that endophytic fungi play an important role in the production plant bioactive metabolites. Moreover, they provide an exploitable basis to increase cryptotanshinone production in S. abrotanoides.

Comparative proteomics analysis of whitetop (Lepidium draba L.) seedlings in response to exogenous glucose.

In this research, a comparative proteomics approach was conducted to understand the physiological processes behind the sulforaphane formation in whitetop seedlings in response to exogenous glucose. Initially, 5-day-old whitetop seedlings were elicited by different concentrations (0, 166, 250, 277, 360 mM) of glucose for 72 h. According to the results, sulforaphane formation was influenced in a dose-dependent manner by glucose, and was maximized with the concentrations of 166 and 250 mM. Consequently, 2-dimensional gel electrophoresis was performed on the 166 mM glucose-elicited seedlings and it was shown that 25 protein spots were differentially expressed between glucose-elicited seedlings and control. Two hypothetical (were down-regulated) and 9 unique proteins (44% and 56% up- and down-regulated, respectively) were identified based on the Mass spectrometry analysis. According to the functional classification of the unique proteins, photosynthetic, chaperone, energy metabolism, signaling and sorting related proteins are marked in response to the glucose elicitation. This is the first report to successfully identify the Abscisic acid receptor PYR1-like and sorting nexin 1 isoform X1 by proteomics technique. In addition, the role of the sorting nexin 1 isoform X1 in the glucose-elicited whitetop seedling is reported for the first time.

Developmental role of phenylalanine-ammonia-lyase (PAL) and cinnamate 4-hydroxylase (C4H) genes during adventitious rooting of Juglans regia L. microshoots.

Phenylalanine-ammonia-lyase and cinnamate-4-hydroxylase play important role in the phenylpropanoid pathway, which produces many biologically important secondary metabolites participating in normal plant development. Flavonol quercetin is the main representant of these compounds that has been identified in numerous Juglans spp. In this survey, the developmental expression patterns of PAL and C4H genes during in vitro rooting of two walnut cultivars 'Sunland' and 'Howard' was examined by RT-PCR. To understand the potential role in rooting, the changing pattern of endogenous content of quercetin was also analyzed by HPLC. The 'Sunland' with better capacity to root had more quercetin content during the "inductive phase" of rooting than 'Howard'. In each cultivar, the level of PAL transcripts showed the same behavior with the changing patterns of quercetin during root formation of microshoots. The positive correlation between the changes of quercetin and PAL-mRNA indicated that PAL gene may have an immediate effect on flavonoid pathway metabolites including quercetin. Although the behavioral change of C4H expression was similar in both cultivars during root formation (with significantly more level for 'Howard'), it was not coincide with the changes of quercerin concentrations. Our results showed that C4H function is important for the normal development, but its transcriptional regulation does not correlate with quercetin as an efficient phenolic compound for walnut rhizogenesis.

Long-term exposure of rapeseed (Brassica napus L.) to ZnO nanoparticles: anatomical and ultrastructural responses.

Rapid development of nanotechnology in recent years has raised concerns about nanoparticle (NPs) release into the environment and its adverse effects on living organisms. The present study is the first comprehensive report on the anatomical and ultrastructural changes of a variety of cells after long-term exposure of plant to NPs or bulk material particles (BPs). Light and electron microscopy revealed some anatomical and ultrastructural modifications of the different types of cell in the root and leaf, induced by both types of treatment. Zinc oxide (ZnO) BPs-induced modifications were surprisingly more than those induced by ZnO NPs. The modifications induced by ZnO BPs or ZnO NPs were almost similar to those induced by excess Zn. Zn content of the root and leaf of both ZnO NPs- and ZnO BPs-treated plants was severely increased, where the increase was greater in the plants treated with ZnO BPs. Overall, these results indicate that the modifications induced by ZnO particles can be attributed, at least partly, to the Zn(2+) dissolution by ZnO particles rather than their absorption by root and their subsequent effects.