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The distinctive characteristics of nanoparticles and their potential applications have been given considerable attention by scientists across different fields, particularly agriculture. However, there has been limited effort to assess the impact of copper nanoparticles (CuNPs) in modulating physiological and biochemical processes in response to salt-induced stress. This study aimed to synthesize CuNPs biologically using Solenostemma argel extract and determine their effects on morphophysiological parameters and antioxidant defense system of barley ( Hordeum vulgare ) under salt stress. The biosynthesized CuNPs were characterized by (UV–vis spectroscopy with Surface Plasmon Resonance at 320 nm, the crystalline nature of the formed NPs was verified via XRD, the FTIR recorded the presence of the functional groups, while TEM was confirmed the shape (spherical) and the sizes (9 to 18 nm) of biosynthesized CuNPs. Seeds of barley plants were grown in plastic pots and exposed to different levels of salt (0, 100 and 200 mM NaCl). Our findings revealed that the supplementation of CuNPs (0, 25 and 50 mg/L) to salinized barley significantly mitigate the negative impacts of salt stress and enhanced the plant growth-related parameters. High salinity level enhanced the oxidative damage by raising the concentrations of osmolytes (soluble protein, soluble sugar, and proline), malondialdehyde (MDA) and hydrogen peroxide (H 2 O 2 ). In addition, increasing the activities of enzymatic antioxidants, total phenol, and flavonoids. Interestingly, exposing CuNPs on salt-stressed plants enhanced the plant-growth characteristics, photosynthetic pigments, and gas exchange parameters. Furthermore, CuNPs counteracted oxidative damage by lowering the accumulation of osmolytes, H 2 O 2 , MDA, total phenol, and flavonoids, while simultaneously enhancing the activities of antioxidant enzymes. In conclusion, the application of biosynthesized CuNPs presents a promising approach and sustainable strategy to enhance plant resistance to salinity stress, surpassing conventional methods in terms of environmental balance.

This research focused on improving the drought tolerance of Dracocephalum moldavica , a plant vulnerable to water stress, by exploring the combined effects of melatonin spray and mycorrhizal fungus Glomus intraradices inoculation. The experiment was designed as a factorial randomized study to evaluate the plant’s morphological, physiological, and phytochemical responses under different drought conditions (100%, 75%, and 50% field capacity). The findings revealed that the combination of melatonin and mycorrhizal inoculation significantly improved the morphological traits of Moldavian balm under drought conditions. Under severe drought (50% field capacity), chlorophyll a and b levels increased by 26.3% and 35.5%, respectively, when both treatments were applied. Stress indicators, including electrolyte leakage and malondialdehyde content, were substantially reduced with the simultaneous application of melatonin and mycorrhizal symbiosis, indicating decreased cellular damage. Moreover, the combined treatment resulted in the highest activities of the antioxidant enzymes catalase and peroxidase, suggesting that these treatments bolster the plant’s oxidative stress defense mechanisms. Additionally, drought stress alone led to an increase in secondary metabolites like phenolic and flavonoid compounds, which were further amplified by the treatments. The study also observed significant alterations in the essential oil composition of the plant. Drought stress increased the levels of α-pinene, 1,8-cineole, and borneol, and these increases were even more pronounced with the combined treatments. Conversely, the levels of geraniol and geranial decreased under drought stress and further with treatment. Overall, this research demonstrates that melatonin and Glomus intraradices inoculation can effectively enhance drought tolerance in Dracocephalum moldavica by improving its physiological characteristics and biochemical composition.

Drought is one of the main limitations impacting the growth, yield, and phytochemical properties of medicinal plants. The present study examined how foliar application of nano-silicon (n-Si) can impact drought tolerance, physiological responses, and levels of bioactive compounds (bio-actives) in the medicinal herb, Lallemantia iberica . Plants were grown in a greenhouse under either well-watered (100% field capacity) or moderate drought stress (50% field capacity) conditions. Drought-stressed plants received a spraying of n-Si (at a concentration of 100 mg L − 1 ) or bulk silicon at a similar concentration, every three days. Compared to control (well-watered) plants, biomass accumulation and photosynthetic efficiency were significantly decreased, along with seed yield and essential oil yields in drought-stressed plants. Application of nano-silicon in drought conditions improved plant water condition, chlorophyll levels, antioxidant enzyme activity, and yield-related traits compared with untreated drought-stressed plants. On the contrary, the responses produced by bulk silicon were much weaker and inconsistent. The improvement in the beneficial effects of nano-silicon resulted from better regulation of water-use efficiency, oxidative stress, and the number of secondary metabolites produced, demonstrating that there were physiological changes instead of isolated responses by individual traits. Results indicate that the beneficial effects of n-Si can lessen the impact of drought stress through the enhancement of physiological stability and the production of phytochemicals from L. iberica in a controlled environment.

This study aims to explore the low phosphorus (P) tolerance of saplings from different Gleditsia sinensis Lam. families. It also seeks to screen for Gleditsia sinensis families with strong low P tolerance and identify key indicators for evaluating their tolerance. This research provides a foundation for the breeding of superior families of Gleditsia sinensis and the study of mechanisms underlying low P tolerance. Using saplings from 30 Gleditsia sinensis families as the research subjects, a sand culture pot experiment was conducted. This study set up low P treatment (0.01 mmol L −1 ) and normal P treatment (1 mmol L −1 ). Twenty-five indicators including growth morphology, biomass, root morphology, and P content were measured. The low P tolerance coefficient was used as the basic data for assessing the low P tolerance of Gleditsia sinensis . The fuzzy comprehensive evaluation method was employed to comprehensively assess the low P tolerance types of Gleditsia sinensis a stepwise regression model was established to identify the key evaluation indicators for low P tolerance. The results indicate that low P stress reduced plant height, stem diameter, and biomass in most Gleditsia sinensis families, but increased the root morphological indicators, root-shoot ratio and PUE of various organs. Principal component analysis transformed the 25 indicators into 6 independent comprehensive indicators, with a cumulative contribution rate of 86.743%. The fuzzy comprehensive evaluation method calculated a comprehensive evaluation value (D value), enabling the screening of Gleditsia sinensis families into low P tolerant and low P sensitive types. Cluster analysis grouped the 30 Gleditsia sinensis families into 4 types. Among them, F13, F10, F9, F18, F15, and F28 were classified as low P tolerant types; F6, F23, F3, F17, F20, F2, F12, F11, F16, F8, F5, F27, F1, and F26 were categorized as intermediate types; F30, F7, F22, F4, F19, F29, F24, F14 and F25 were considered low P sensitive types, and F21 was classified as extremely low P sensitive types. The stepwise regression analysis identified the indicators stem diameter, total root volume, shoot dry weight, total root projection area, and leaf P content as the key factors for discriminating the low P tolerance of Gleditsia sinensis . The regression model is as follows: D=-0.005 + 0.323 stem diameter *+0.154 * total root volume + 0.196* shoot dry weight + 0.139* total root projection area − 0.112* leaf P content. In summary, low P stress inhibited the growth of Gleditsia sinensis saplings, but it increased the root morphological indicators, root-shoot ratio and PUE of various organs to cope with low P environments. The screening identified F13, F10, F9, F18, F15, and F28 as low P tolerant Gleditsia sinensis families. The evaluation indicators for low P tolerance in Gleditsia sinensis were identified as stem diameter, total root volume, shoot dry weight, total root projection area and leaf P content.

Background Glechoma longituba , recognized as a medicinal plant, provides valuable pharmaceutical raw materials for treating various diseases. Saline-alkali stress may effectively enhance the medicinal quality of G. longituba by promoting the synthesis of secondary metabolites. To investigate the changes in the primary medicinal components of G. longituba under saline-alkali stress and improve the quality of medicinal materials, Na 2 CO 3 was applied to induce short-term stress under different conditions and the biomass, physiologically active substances and primary medicinal components of G. longituba were measured in this study. Results Under alkaline salt stress, the activities of catalase (CAT), superoxide dismutase (SOD), peroxidase (POD), and ascorbate peroxidase (APX) were elevated in G. longituba , accompanied by increased accumulation of proline (Pro) and malondialdehyde (MDA). Furthermore, analysis of the medicinal constituents revealed that G. longituba produced the highest levels of soluble sugars, flavonoids, ursolic acid, and oleanolic acid under 0.6% Na 2 CO 3 stress for 48 h, 0.2% Na 2 CO 3 stress for 72 h, 0.4% Na 2 CO 3 stress for 12 h, and 0.4% Na 2 CO 3 stress for 8 h, respectively. Conclusions Short-term Na 2 CO 3 stress enhances the synthesis of medicinal components in G. longituba . By manipulating stress conditions, the production of various medicinal substances could be optimized. This approach may serve as a basis for the targeted cultivation of G. longituba , offering potential applications in the treatment of diverse diseases.

Clinopodium nepeta (L.) Kuntze is a medicinal and aromatic species of the Lamiaceae family, rich in phenolic compounds; however, studies regarding its in vitro culture, growth regulation properties and secondary metabolism remain limited. The present study investigated the effects of three adenine-type cytokinins: 6-benzylaminopurine (BAP), meta-topolin (m-TOP), and BAP riboside (r-BAP), applied at concentrations of 0.5, 1.0, 2.0 or 4.0 mg L−1, on shoot proliferation, biomass accumulation, phenolic profile, and antioxidant activity in C. nepeta shoot cultures. The phenolic constituents of hydromethanolic shoot extracts were subjected to qualitative profiling using UHPLC–DAD–ESI–MS, while antioxidant potential was evaluated using spectrophotometric assays (DPPH, ABTS, FRAP). All tested cytokinins stimulated shoot proliferation and biomass growth compared with the control; of these, m-TOP demonstrated the most pronounced positive effect, characterized by high multiplication rate and improved shoot morphology. UHPLC–DAD–ESI–MS analysis revealed the presence of numerous caffeic acid derivatives including rosmarinic acid, chlorogenic acids, salvianolic acid derivatives, and flavonoid glycosides; their accumulation was strongly influenced by cytokinin type and concentration. Notably, rosmarinic acid, the dominant phenolic constituent in the treated shoots, reached 23.28 mg g−1 DW under m-TOP treatment, i.e., an approximate 20-fold increase compared with the control. The extracts from shoots cultured on cytokinin-supplemented media exhibited enhanced antioxidant activity, which correlated with increased phenolic content. These relationships were confirmed by principal component analysis (PCA). Hence, C. nepeta shoot cultures represent an efficient in vitro system for biomass production and phenolic compound biosynthesis, and the selection of cytokinin type is a critical factor modulating both morphogenetic and metabolic responses.

Silver nanoparticles (AgNPs) are increasingly recognized as active elicitors capable of modulating plant growth and metabolism. This study evaluated the effects of AgNPs (1.0–50.0 mg/L) on in vitro shoot cultures of Clinopodium nepeta subsp. spruneri (Boiss.) Bartolucci & F. Conti (Lamiaceae). AgNP exposure significantly reduced shoot length, leaf and node numbers, and completely suppressed root formation, while biomass accumulation increased in a concentration-dependent manner. High-performance liquid chromatography (HPLC) identified rosmarinic acid (RA) as the major phenolic, with levels rising 2.5- to 4.5-fold in treated shoots compared with control and wild plants. The maximum RA content, total phenolic concentration, antioxidant activity (DPPH and FRAP), and acetylcholinesterase (AChE) inhibition were observed at 20.0 mg/L AgNP. In contrast, the highest flavonoid content was recorded in the wild plant. Gas chromatography–mass spectrometry (GC-MS) analysis revealed that isoeugenol dominated in AgNP-treated cultures, while menthalactone was the principal constituent in both control and wild samples. These findings demonstrate that AgNPs substantially alter the metabolic profile of C. nepeta subsp. spruneri , highlighting their potential as elicitors for enhancing phenolic accumulation, antioxidant activity, and enzyme inhibition under in vitro conditions.

This study examined the dual effects of aluminum oxide nanoparticles (Al O NPs) on Dracocephalum kotschyi Boiss, focusing on physiological, biochemical, and antioxidant defense responses. Moreover, the root and shoot length, biomass production, uptake, and translocation of Al were measured in the studied plants under different doses of Al O NPs (0, 50, 100, 1000, and 2500 ppm). Field Emission Scanning Electron Microscopy (FE-SEM) analysis revealed Al O NPs clusters on the plant root surface. The Al O NPs at 50 ppm promoted the root and shoot dry weight (65.6 and 47.9%, respectively) and the root elongation (21.9%), whereas they decreased the root biomass at 1000 ppm. The amount of chlorophylls (Chl-a and Chl-b) and carotenoids in the fresh leaves of D. kotschyi plants grown under Al O NPs treatment at a dose of 1000 ppm increased compared to the untreated control plants. An increase in total soluble sugar and protein levels at all doses and proline content at 100-2500 ppm was observed in plant exposed to Al O NPs. In response to the toxic effects of Al O NPs, the treated plants stimulated the enzymatic defense system with changes in superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and ascorbate peroxidase (APX) activities. In addition, D. kotschyi plants activated non-enzymatic antioxidants to detoxify the impacts of Al O NPs by increasing the total level of flavonoid, saponin, and iridoid as reactive oxygen species (ROS)-scavenging compounds. Accordingly, the stimulatory and inhibitory impacts of Al O NPs in the treated plants depended on the applied doses and their physicochemical properties; however, these responses may differ in different plants.

Harpagide and its derivatives have valuable medicinal properties, such as anti-inflammatory, analgesic and potential antirheumatic effects. There is the demand for searching plant species containing these iridoids or developing biotechnological methods to obtain the compounds. The present study investigated the effects of methyl jasmonate (MeJa, 50 μM), ethephon (Eth, 50 μM) and L-phenylalanine (L-Phe, 2.4 g/L of medium), added to previously selected variant of Murashige and Skoog medium (supplemented with plant growth regulators: 6-benzylaminopurine 1.0 mg/L, α-naphthaleneacetic acid 0.5 mg/L, gibberellic acid 0.25 mg/L) on the accumulation of harpagide and 8-O-acetyl-harpagide in Melittis melissophyllum L. agitated shoot cultures. Plant material was harvested 2 and 8 days after the supplementation. Iridoids were quantitatively analyzed by the UPLC-MS/MS method in extracts from the biomass and the culture medium. It was found that all of the variants caused an increase in the accumulation of harpagide. In the biomass harvested after 2 days, the highest harpagide content of 247.3 mg/100 g DW was found for variant F (L-Phe and Eth), and the highest 8-O-acetyl-harpagide content of 138 mg/100 g DW for variant E (L-Phe and MeJa). After 8 days, in some variants, a portion of the metabolites was released into the culture medium. Considering the total amount of the compounds (in the biomass and medium), the highest accumulation of harpagide, amounting to 619 mg/100 g DW, was found in variant F, and the highest amount of 8-O-acetyl-harpagide, of 255.4 mg/100 g DW, was found in variant H (L-Phe, MeJa, Eth) when harvested on the 8th day. These amounts were, respectively, 24.7 and 4.8 times higher than in the control culture, and were, respectively, 15 and 6.7 times higher than in the leaves of the soil-grown plant. The total amount of the two iridoids was highest for variant F (0.78% DW) and variant H (0.68% DW) when harvested on the 8th day. The results indicate that the agitated shoot cultures of M. melissophyllum can be a rich source of harpagide and 8-O-acetyl-harpagide, having a potential practical application. To the best of our knowledge we present for the first time the results of the quantitative UPLC-MS/MS analysis of harpagide and 8-O-acetyl-harpagide in M. melissophyllum shoot cultures and the enhancement of their accumulation by means of medium supplementation with elicitors and precursor.

Background Dracocephalum kotschyi Boiss., a perennial medicinal and aromatic plant endemic to Iran, is renowned for its diverse biological and medicinal properties. This study aimed to evaluate the influence of foliar application of selenium nanoparticles (SeNPs) and sodium selenite (SSe) on the growth, yield, physiological and biochemical characteristics, and essential oil (EO) of D. kotschyi . The experiment included seven treatments: a control (distilled water) and a foliar spray of 50, 100, and 200 mg L −1 of both SeNPs and SSe separately. Results SeNPs were more effective than SSe in improving plant growth and yield. Specifically, a 200 mg L −1 SeNPs treatment led to the highest plant height, fresh weight of the plant, dry weight of the leaf, and relative water content. Elevated Se concentrations were associated with a decrease in chlorophyll levels. Both SeNPs and SSe treatments increased hydrogen peroxide levels, and catalase and ascorbate peroxidase activity. The phenylalanine ammonia-lyase activity increased with SeNPs up to 50 mg L −1 and SSe up to 100 mg L −1 . Application of 200 mg L −1 SSe resulted in the highest total phenol content and antioxidant activity. Although spraying 100 mg L −1 SeNPs led to the highest EO content, the EO yield peaked with 200 mg L −1 SeNPs. Notably, applying 200 mg L −1 SeNPs reduced the level of neral, while 200 mg L −1 SSe decreased both geranial and neral levels. Conversely, levels of α -pinene and geranyl acetate increased with Se application. Conclusions The results revealed that SeNPs were more effective than the SSe in improving plant growth and yield. Overall, the 200 mg L −1 SeNPs treatment was the most effective in improving the growth, yield attributes, and EO yield of D. kotschyi.