The Influence of Methyl Jasmonate on Expression Patterns of Rosmarinic Acid Biosynthesis Genes, and Phenolic Compounds in Different Species of Salvia subg. Perovskia Kar L.

Salvia yangii B.T. Drew and Salvia abrotanoides Kar are two important fragrant and medicinal plants that belong to the subgenus Perovskia. These plants have therapeutic benefits due to their high rosmarinic acid (RA) content. However, the molecular mechanisms behind RA generation in two species of Salvia plants are still poorly understood. As a first report, the objectives of the present research were to determine the effects of methyl jasmonate (MeJA) on the rosmarinic acid (RA), total flavonoid and phenolic contents (TFC and TPC), and changes in the expression of key genes involved in their biosynthesis (phenylalanine ammonia lyase (PAL), 4-coumarate-CoA ligase (4CL), and rosmarinic acid synthase (RAS)). The results of High-performance liquid chromatography (HPLC) analysis indicated that MeJA significantly increased RA content in S. yungii and S. abrotanoides species (to 82 and 67 mg/g DW, respectively) by 1.66- and 1.54-fold compared with untreated plants. After 24 h, leaves of Salvia yangii and Salvia abrotanoides species treated with 150 M MeJA had the greatest TPC and TFC (80 and 42 mg TAE/g DW, and 28.11 and 15.14 mg QUE/g DW, respectively), which was in line with the patterns of gene expression investigated. Our findings showed that MeJA dosages considerably enhanced the RA, TPC, and TFC contents in both species compared with the control treatment. Since increased numbers of transcripts for PAL, 4CL, and RAS were also detected, the effects of MeJA are probably caused by the activation of genes involved in the phenylpropanoid pathway.

Methyl Jasmonate Induces Genes Involved in Linalool Accumulation and Increases the Content of Phenolics in Two Iranian Coriander (Coriandrum sativum L.) Ecotypes.

The medicinal herb coriander (Coriandrum sativum L.), with a high linalool (LIN) content, is widely recognized for its therapeutic benefits. As a novel report, the goals of this study were to determine how methyl jasmonate (MeJA) affects total phenolic content (TPC), LIN content, flavonoid content (TFC), and changes in gene expression involved in the linalool biosynthesis pathway (CsγTRPS and CsLINS). Our findings showed that, in comparison to the control samples, MeJA treatment substantially enhanced the TPC, LIN, and TFC content in both ecotypes. Additionally, for both Iranian coriander ecotypes, treatment-induced increases in CsγTRPS and CsLINS expression were connected to LIN accumulation in all treatments. A 24 h treatment with 150 µM MeJA substantially increased the LIN content in the Mashhad and Zanjan ecotypes, which was between 1.48 and 1.69 times greater than that in untreated plants, according to gas chromatography-mass spectrometry (GC-MS) analysis. Our findings demonstrated that MeJA significantly affects the accumulation of LIN, TPC, and TFC in Iranian C. sativum treated with MeJA, which is likely the consequence of gene activation from the monoterpene biosynthesis pathway. Our discoveries have improved the understanding of the molecular mechanisms behind LIN synthesis in coriander plants.

Ion content, antioxidant enzyme activity and transcriptional response under salt stress and recovery condition in the halophyte grass Aeluropus littoralis.

OBJECTIVE: In contrast to glycophytes, halophyte plants have evolved unique morphological and physiological mechanisms to deal with abiotic stress. This study presents the physiological responses of Aeluropus littoralis, a halophyte grass, to salt stress and recovery conditions on the molecular level. RESULTS: Elemental analysis showed that Na+ concentration increased in the analyzed tissue during salt stress application, and declined at recovery condition. With the exception of root tissue, comparable trends of K+, Ca2+, and Mg2+ concentrations were observed (decreased during salt stress, increased during recovery). Salinity led to an increase in total chlorophyll (Chl), Chl a, and carotenoids content, while Chl b content decreased. The level of the proline amino acid associated with drought and salt stress was increased. Here APX, POD, and SOD activity were strongly detectable in roots and reduced later under recovery conditions. RT-qPCR revealed up-regulation of antioxidant genes at S1 and S3 in the root but down-regulation in recovery conditions. This study found a significant halophyte index for understanding the processes of salinity tolerance in A. littoralis. These findings may provide insight into the role of antioxidant enzymes during salt stress and the mechanism underlying the plant's response to stress.

Phytochemical, antioxidant, enzyme activity and antifungal properties of Satureja khuzistanica in vitro and in vivo explants stimulated by some chemical elicitors.

Context: Satureja khuzistanica Jamzad. (Lamiaceae), is known for its antifungal and antioxidant compounds, especially rosmarinic acid (RA).Objective: The study examines the effect of elicitors on RA production and phytochemical properties of S. khuzistanica.Materials and methods: In vitro plants were treated with methyl jasmonate (MeJA) and multi-walled carbon nanotubes (MWCNTs). In vivo plants were treated with MWCNTs and salicylic acid (SA). RA was measured by HPLC. Catalase (CAT), guaiacol peroxidase (POD) and ascorbate peroxidase (APX) were quantified. DPPH and ß-carotene were assayed in in vivo extracts. The antifungal effects of extracts were evaluated against Fusarium solani K (FsK).Results: The highest RA contents of in vitro plants were 50 mg/L MeJA (140.99 mg/g DW) and 250 mg/L MWCNTs (140.49 mg/g DW). The highest in vivo were 24 h MWCNTs (7.13 mg/g DW) and 72 h SA (9.12 mg/g DW). The maximum POD and APX activities were at 100 mg/L MeJA (5 and 4 mg protein, respectively). CAT had the highest activities at 50 mg/L MeJA (2 mg protein). DPPH and ß-carotene showed 50% and 80% inhibition, respectively. The FsK aggregation was the lowest for in vitro extract in number of conidia [1.82 × 1010], fresh weight (6.51 g) and dry weight (0.21 g) that proved RA inhibitory effects. The callus reduces FsK growth diameter to 2.75 on the 5th day.Discussion and conclusions: Application of MeJA, SA, and MWCNTSs could increase RA in S. khuzistanica and highlighted potential characteristics in pharmaceutical and antifungal effects.

Exploiting Differential Gene Expression to Discover Ionic and Osmotic-Associated Transcripts in the Halophyte Grass Aeluropus littoralis.

BACKGROUND: Salinity as a most significant environmental challenges affects the growth and productivity of plants worldwide. In this study, the ionic and iso-osmotic effects of salt stress were investigated in Aeluropus littoralis L., a halophyte grass species from Poaceae family, by cDNA-amplified fragment length polymorphism (cDNA-AFLP) technique. To dissect the two different effects (ionic and osmotic) exerted by salt stress, various ionic agents including 200 and 400 mM sodium chloride (NaCl), 200 and 400 mM potassium chloride (KCl) as well as 280 and 406 gl- 1 (- 0.9 and - 1.4 MPa) polyethylene glycol 6000 (PEG) as their iso-osmotic concentrations were applied. RESULTS: Application of KCl and PEG significantly reduced the fresh weight (FW) of A. littoralis seedlings compared to control while NaCl treatment markedly enhanced the FW. At the transcriptome level, different observations of changes in gene expression have been made in response of A. littoralis to ionic and osmotic stresses. Out of 69 transcript derived fragments (TDFs), 42 TDFs belong to 9 different groups of genes involved in metabolism (11.6%), transcription (10.2%), ribosomal protein (8.7%), protein binding (8.7%) transporter (5.8%), translation (5.8%), signal transduction (4.3%), nucleosome assembly protein (2.9%) and catabolism (2.9%). The 44 and 28 percent of transcripts were expressed under ionic stress (NaCl-specific and KCl-specific) and osmotic stress (common with NaCl, KCl and PEG), respectively which indicating a greater response of plants to ionic stress than osmotic stress. Expression pattern of eight candidate TDFs including; SYP81, CAND1, KATN, ISB1, SAMDC, GLY1, HAK18 and ZF30 was evaluated by RT-qPCR at high salinity levels and recovery condition. CONCLUSION: Differential regulation of these TDFs was observed in root and shoot which confirm their role in salt stress tolerance and provide initial insights into the transcriptome of A. littoralis. Expression pattern of ionic and osmotic-related TDFs at A. littoralis can be taken as an indication of their functional relevance at different salt and drought stresses.

The Effect of Food-Simulating Agents on the Bond Strength of Hard Chairside Reline Materials to Denture Base Resin.

PURPOSE: To investigate the influence of food-simulating agents on the shear bond strength between direct hard liners and denture base acrylic resin. In addition, mode of failure was evaluated. MATERIALS AND METHODS: One hundred fifty cylindrical columns of denture base resin were fabricated and bonded to three types of hard reline materials (Hard GC Reline, Tokuyama Rebase II Fast, TDV Cold Liner Rebase). Specimens of each reline material were divided into five groups (n = 10) to undergo 12-day immersion in distilled water, 0.02 N citric acid aqueous solution, heptane, and 40% ethanol/water solution at 37°C. The control group was not immersed in any solution. The shear bond strength test was performed, and the failure mode was determined. Statistics were analyzed with two-way ANOVA and chi-square test (α = 0.05). RESULTS: Significant interaction was found between the hard liners and food simulating agents (p < 0.001). The shear bond strength of Tokuyama in 40% ethanol and TDV in heptane decreased significantly (p = 0.001, p < 0.001 respectively); however, none of the solutions could significantly affect the shear bond strength of Hard GC Reline (p = 0.208). The mixed failure mode occurred more frequently in Hard GC Reline compared with the other liners (p < 0.001) and was predominant in specimens with higher bond strength values (p = 0.012). CONCLUSIONS: Food simulating agents did not adversely affect the shear bond strength of Hard GC Reline; however, ethanol and heptane decreased the bond strength of Tokuyama and TDV, respectively. These findings may provide support to dentists to recommend restricted consumption of some foods and beverages for patients who have to use dentures relined with certain hard liners.

Bond strength of hard direct reline materials to heat-cured acrylic denture base after immersion in denture cleansers.

BACKGROUND: Immersion-type denture cleansers are commonly used for denture hygiene maintenance. Hence, it is crucial to investigate the effect of denture cleansing solutions on bond strength between direct reline materials and denture base resin. AIMS: This in vitro study aimed to determine the effect of denture cleansers on bond strength between direct hard reline materials and denture base resin. MATERIALS AND METHODS: Cylindrical columns of hard-liners (Hard GC Reline, TDV Cold Liner Rebase, Tokuyama Rebase II Fast) were bonded to heat-polymerized denture base resin. A total of fifty specimens were fabricated for each reline material and divided into five groups (n = 10): Group I (control): No solution was used; Group II: Specimens were stored in distilled water for 60 days; Groups III, IV, and V: Specimens were stored in distilled water for 60 days with daily immersion in either sodium hypochlorite, calgon + sodium hypochlorite, or dentipur tablet for 5 min. The shear bond strength was examined at a cross-head speed of 1 mm/min. Failure mode was evaluated by stereomicroscope. STATISTICAL ANALYSIS: Data were analyzed by two-way ANOVA and Chi-square test (α=0.05). RESULTS: The results showed no significant interaction between the direct hard-liners and denture cleansers (P = 0.119). Hard GC Reline had the highest bond strength, followed by Tokuyama Rebase II Fast, and then, TDV Cold Liner Rebase. No significant difference existed in bond strength between samples immersed in water and cleansers or between the cleansers themselves. Hard GC Reline had more mixed failure mode compared to TDV Cold Liner Rebase and Tokuyama Rebase II Fast. There was a significant correlation between mixed mode of failure and higher values of bond strength (P = 0.008). CONCLUSIONS: Within the limitations of the present study, denture cleansing solutions could not significantly influence the bond strength between hard direct liners and denture base resin.

Three-Component Reaction of Benzothiazole, Acetylenic Esters, Phenoles; Synthesis of Dialkyl 2-benzo[d]thiazol Derivatives Under Grinding.

AIM AND OBJECTIVE: Due to biological activity of a significant number of compounds containing benzothiazole ring system and in continuation of our interest in developing new environmentally benign methods for the synthesis of heterocyclic compounds by MCRs in this study we performed an efficient three-component from benzothiazole, acetylenic esters and hydroxyl aromatics compounds to synthesize of 2- benzothiazole derivatives in high yield. MATERIALS AND METHODS: IR spectra were recorded using an FTIR apparatus. Melting points measured on an Electrothermal 9100 apparatus. Spectra were obtained in solution of CDCl3 using TMS as internal standard. Elemental analyses were carried out using a Heracus CHN-O- Rapid analyzer. A mixture of benzothiazole, dimethyl acetylenedicarboxylate and phenol were placed in a mortar. The mixture was ground with a mortar and pestle at room temperature for 12 min. After completion of the reaction, as indicated by TLC (ethyl acetate: n-hexane, 1: 3), the solvent was distilled off under reduced pressure and the residue was crystallized from diethyl ether. RESULTS: Treatment of benzothiazole and dialkyl acetylenedicarboxylates in presence of resorcinol and ß-naphthol led to products 4 (dialkyl (E)-2-(2-(2-hydroxyphenyl)benzo[d]thiazol-3(2H)-yl-3- methylbut-2-enedioate), while we observed two isomer (Z) and (E) configurations (major and minor) in nearly 70:30 ratio when the reaction was repeated in presence of 8-hydroxy quinolone and DMAD. Also when we examined 2-nitro phenol, 4-nitrophenol, and 4-hydroxy quinoline, only one product 5 was obtained. This indicates that the reaction proceeds efficiently with electron-releasing substituted phenols. CONCLUSION: The reaction between benzothiazole and dialkyl acetylenedicarboxylates in the presence of some phenols without electron-withdraw substitution, presents a novel, one-pot, clean, convenient, simple and inexpensive approach into the synthesis of 2-benzothiazole derivatives of potential synthetic and pharmacologically interest. This procedure carries significant advantages because of the minimization of labor, time, and cost.

Green approach to stereoselective synthesis of benzo[d]chromeno[3,4- h]oxathiazonine derivatives via MCRs in water: a combined experimental and DFT study.

(7S, 14S, 16R)- dialkyl 6-oxo-6,7,13,14-tetrahydro-7,14-methanobenzo[d]chromeno[3,4- h][1,6,3]oxathiazo-nine-14,16-dicarboxylates 4 and (S)-methyl 2-(2-methoxy-2-oxoethyl)-2,3- dihydrobenzo[d]thiazole-2-carboxylates 5 were readily synthesized in a ratio 3:1 and moderated yield by multicomponent reactions of 4-hydroxycoumarin and acetylenic esters with benzothiazole without using a catalyst. Also, The GIAO/DFT approach at the B3LYP/6-31G** level of theory was used to calculate the (1)H, (15)N, (17)O and (13)C NMR chemical shifts of product 4b. These computations are performed on the basis of X-ray structural data which are collected at 120(2) K temperature. In order to take into account intermolecular hydrogen bonds and the van der Waals effects, two different sizes of clusters (two model of trimeric and pentameric clusters) have been considered. A comparison between the experimental (Exp.) and calculated (Cal.) (1)H and (13)C NMR chemical shifts may reveal that the solution contains monomer, trimer1, trimer2, and pentamer models.

Evaluation of proliferation activity in dysplastic and nondysplastic oral lichen planus through the analysis of argyrophilic nucleolar organizer regions.

Lichen planus is a mucocutaneous disease, and its potential for malignant transformation is a subject of controversy. We found dysplastic changes in slide review of about 11% of oral lichen planus (OLP) in our previous research. Dysplastic changes can be an initial phase for carcinogenesis. The question is whether it is possible to detect early malignant changes in OLP through the evaluation of proliferation activity. The aim of this study was to compare the cell proliferation activity in dysplastic and nondysplastic oral lichen planus using the quantitative and qualitative analysis of argyrophilic nucleolar organizer region (AgNOR) and to compare candidal infection in these 2 groups. In this cross-sectional, observational, and analytical study, all 14 cases of confirmed dysplastic OLPs and 17 nondysplastic OLPs were stained with an AgNOR technique. Analysis of argyrophilic nucleolar organizer region dots were counted in 100 cells of basal and parabasal layers, and proliferation index (cells with ≥ 5 dots in nucleus) and variations in size of AgNOR dots were also evaluated. Periodic acid-Schiff staining was performed for detection of candidal infection. Mean AgNOR count in dysplastic OLP (2.39 ± 0.2) was more than nondysplastic OLP (2.27 ± 0.59). Candidiasis was found in 14.3% of dysplastic OLP and in 12.5% of nondysplastic OLP. There was no statistically significant difference between the 2 groups in mean AgNOR count (P = 0.62), variation in size of AgNOR dots (P = 1), proliferation index (P = 0.53), and candidiasis (P = 1). The current study showed slight difference in proliferation rate and candidal infection between dysplastic and nondysplastic OLP.