Proteomic and physiological analyses to elucidate nitric oxide-mediated adaptive responses of barley under cadmium stress.

Nitric oxide (NO) is known to induce plant resistance for several environmental stresses. The protective roles of NO in cadmium (Cd) toxicity have been well documented for various plant species; nevertheless, little information is available about its molecular regulation in improving Cd tolerance of barley plants. Therefore, we combined a comparative proteomics with physiological analyses to evaluate the potential roles of NO in alleviating Cd stress (50 µM) in barley (Hordeum vulgare L.) seedlings. Exogenous application of NO donor sodium nitroprusside (SNP, 100 µM) decreased the Cd-mediated seedling growth inhibition. This observation was supported by the reduction of lipid peroxidation as well as the improvement of chlorophyll content and inhibition of hydrogen peroxide accumulation. Activities of the superoxide dismutase and guaiacol peroxidase were reduced following the application of SNP, while ascorbate peroxidase activity was enhanced. In this study, a total of 34 proteins were significantly regulated by NO in the leaves under Cd stress using a gel-based proteomic approach. The proteomic analysis showed that several pathways were noticeably influenced by NO including photosynthesis and carbohydrate metabolism, protein metabolism, energy metabolism, stress defense, and signal transduction. These results provide new evidence that NO induce photosynthesis and energy metabolism which may enhance Cd tolerance in barley seedlings. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-022-01214-3.

Growth responses and essential oil profile of Salvia officinalis L. Influenced by water deficit and various nutrient sources in the greenhouse.

Salvia officinalis L. is a medicinal plant extensively used in foods, traditional medicine, and the pharmacological industry. In the current study, the effects of different irrigation regimes [irrigation after 70 ± 5 (regular), 105 ± 5 (moderate drought stress), and 140 ± 5 (severe drought stress) mm evaporation] and nutrient sources (control, NPK, farmyard manure, foliar fertilizer, and hydrogel) were investigated on the growth parameters and essential oil (EO) components of S. officinalis in the greenhouse. The plants were harvested two times. The regular irrigation treatment had the most significant effect on plant height (51 cm), fresh and dry herb weight (51.5 and 18.1 g plant-1), and fresh and dry leaf weight (40.1 and 13.1 g plant-1). The highest amount of EO was observed after moderate drought stress (1.48%). The NPK treatment had the greatest effect on plant height (40 cm), branch number (19 per plant), fresh and dry herb weight (53.4 and 18.9 g plant-1), fresh and dry leaf weight (41.2 and 13.6 g plant-1), and EO content (1.67%). The 1st cutting was superior in EO amount, while the 2nd cutting had a high agronomic yield. α-Thujone (from 21.6 to 34.2%) was identified as the predominant compound. Additionally, the content of α-thujone in the 2nd cutting was higher after moderate drought stress, NPK, and hydrogel treatments. Moreover, 1,8-cineole, ß-thujone, camphene, α-pinene, α-humulene, viridiflorol, borneol, and bornyl acetate were the other main compounds. As a general result, regular irrigation and NPK treatments improved the agronomic yield of S. officinalis. The plants under drought stress produced high amounts of EO. The farmyard manure also improved plant yield by providing a part of the plant's nutritional needs. Therefore, it could be concluded that it is crucial to determine the effects of limited water availability and various nutrient sources on yield and chemical compositions for medicinal and aromatic plant growth.

Proteomic analysis reveals the role of exogenous cysteine in alleviating chromium stress in maize seedlings.

Cysteine (Cys) is incorporated into several compounds which are involved in detoxification of heavy metals. It is evident from recent studies that Cys is effective in alleviating the toxicity of heavy metals. Nevertheless, little is known about the Cys-mediated alleviation of chromium (Cr) toxicity. In our study, the impacts of exogenous Cys on Cr-stressed maize (Zea mays L.) were examined by using physiological and proteomic analyses. The results showed that Cr (100 µM) increased the accumulation of hydrogen peroxide, decreased cell viability, enhanced lipid peroxidation and consequently inhibited plant growth. The application of Cys (500 µM) attenuated the adverse effects of Cr on seedling growth. Cys supplementation to Cr treated plants decreased Cr accumulation in the shoots and increased Cr accumulation in roots. Cys treatment also modulated the activities of antioxidant enzymes and increased endogenous Cys content. Sixty proteins in root tissue were significantly affected by exogenous Cys under Cr stress using two-dimensional electrophoresis. Forty-six differentially expressed proteins were successfully identified by MALDI-TOF/TOF mass spectrometry. These differentially expressed proteins were involved in various biological pathways such as stress response (41.3%), energy and carbohydrate metabolism (21.7%), protein metabolism (6.5%), amino acid metabolism (6.5%), and others of unknown functions. The defense response-related proteins including glutathione peroxidase, glutathione S-transferases, pathogenesis-related proteins, glyoxalases and superoxide dismutase were differently regulated by Cys suggesting their roles in the Cys-mediated Cr tolerance.

Proteomic analysis of the anticancer effect of various extracts of endemicThermopsisturcica in human cervical cancer cells

Background/aim: Thermopsisturcica is a perennial species endemic to Turkey and different extracts of T. turcica have an antiproliferative effect on cancer cells, but there has not been any report on HeLa (human cervical cancer) cells. Materials and methods: To get a better understanding of the molecular mechanism of anticancer activity of methanolic extracts of leaves (LE) and flowers (FE) of T. turcica, we employed 2-DE-based proteomics to explore the proteins involved in anticancer activity in HeLa cells. Results: T. turcica extracts showed a potent cytotoxic effect on HeLa cells with the IC50 values of 1.75 mg/mL for LE and 3.25 mg/mL for FE. The induction of apoptosis by LE and FE was also consistent with increased expression of caspase mRNAs and DNA fragmentation. In terms of the proteomic approach, 27 differentially expressed proteins were detected and identified through MALDI-TOF/TOF mass spectrometry. These altered proteins were involved in cytoskeleton organization and movement, protein folding, proteolysis and translation, cell cycle and proliferation, signal transduction, cell redox homeostasis, and metabolism. Conclusion: Up-regulation of protein disulfide isomerases and down-regulation of Rho GDP-dissociation inhibitor, heterogeneous nuclear ribonucleoproteins, and heat shock proteins may contribute to the induction of apoptosis and arresting of the cell cycle in HeLa cells.

Interactive effects of sulfur and chromium on antioxidative defense systems and BnMP1 gene expression in canola (Brassica napus L.) cultivars differing in Cr(VI) tolerance.

Plants suffer with combined stress of sulfur (S) deficiency and hexavalent chromium [Cr(VI)] in soils. There are a few reports on the interactive effects of S-deficiency and Cr(VI) stress. Therefore, the interactions between S nutrition and Cr(VI) stress were investigated in hydroponically grown canola (Brassica napus L.) cultivars differing in Cr(VI) tolerance. The relatively Cr(VI)-tolerant (NK Petrol) and Cr(VI)-susceptible (Sary) cultivars were grown in S-sufficient nutrient solution and then exposed to variable S concentrations [deficient (0 mM S, -S) and sufficient (1 mM S, +S)]. The seedlings were then exposed to 100 µM Cr(VI) for 3 days. S-deficiency (-S/-Cr) and combined stress (-S/+Cr) caused a significant decrease in growth parameters of Sary than NK Petrol (P < 0.05). In -S/+Cr treatment, Cr accumulation in Sary was significantly higher than NK Petrol. The higher level of Cr in Sary increased lipid peroxidation and decreased chlorophyll content. The activities of antioxidant enzymes and cysteine content were significantly higher in NK Petrol than in Sary under combined stress. The levels of ascorbate (AsA) and glutathione (GSH) were significantly decreased by S deficiency. The expression level of metallothionein gene (BnMP1) in the tolerant NK Petrol was increased by -S/+Cr treatment. However, expression level of BnMP1 gene in the susceptible Sary was enhanced by +S/+Cr treatment. This result suggests metallothionein (MT) may be involved in Cr(VI) tolerance under S-deficient condition. In conclusion, S nutrition influenced Cr accumulation and enhanced tolerance caused by a positive effect on defense systems and gene expression.

Proteomic and biochemical responses of canola (Brassica napus L.) exposed to salinity stress and exogenous lipoic acid.

To evaluate the mitigating effects of exogenous lipoic acid (LA) on NaCl toxicity, proteomic, biochemical and physiological changes were investigated in the leaves of canola (Brassica napus L.) seedlings. Salinity stress decreased the growth parameters and contents of ascorbate (AsA) and glutathione (GSH), and increased the contents of malondialdehyde (MDA), proline, cysteine and the activities of antioxidant enzymes such as superoxide dismutase (SOD), guaiacol peroxidase (POD), catalase (CAT) and ascorbate peroxidase (APX). The foliar application of LA alleviated the toxic effects of salinity stress on canola seedlings and notably decreased MDA content and increased growth parameters, cysteine content, and activities of CAT and POD. In the proteomic analyses, total proteins from the leaves of control, LA, NaCl and NaCl+LA treated-seedlings were separated using two-dimensional gel electrophoresis (2-DE). A total of 28 proteins were differentially expressed. Of these, 21 proteins were successfully identified by MALDI-TOF/TOF MS. These proteins had functions related to photosynthesis, stress defense, energy metabolism, signal transduction, protein folding and stabilization indicating that LA might play important roles in salinity through the regulation of photosynthesis, stress defense and signal transduction related proteins. The proteomic findings have provided new insight to reveal the effect of LA on salinity stress for the first time.

Cytotoxic effects of nasal buserelin on nasal mucosal tissue in rabbits.

To investigate the cytotoxic effects of nasal buserelin on rabbit nasal mucosal tissue, twenty-four female rabbits were studied prospectively. The rabbits were divided into 4 groups including 6 rabbits. The rabbits' left noses were included in the all study groups: 150 µg/puff/day of buserelin acetate was administered topically twice daily during 21, 42 and 63 days. Saline was administered topically twice daily to the left nasal cavity in the control group. The nasal septal mucosal stripe tissue was carefully removed from underlaying cartilage after sedation. HE staining, Masson's trichrome, toluidine blue and TUNEL staining were used to evaluate mucosal changes. Each preparation was investigated via apoptotic cells, and they were accounted. Kruskal-Wallis test was used to evaluate nonparametric comparison of apoptotic cells. Mononuclear cells have been raised in the sub-epithelial connective tissue, nucleuses of epithelial cells in the apical region were pyknotic, and apoptotic cells were determined on 21-day group. In the 42-day group, nasal epithelial tissue was similar to 21-day group and epithelial cells including pyknotic nucleus were present in this group, too. In the 63-day group, epithelial cells were light colored. Venous sinuses in the sub-epithelial connective tissue were wide but not congested and not raised collagen filaments. In the intra-epithelial tissue, some of cells were TUNEL (+). Apoptotic cells were fewer in the control group according to 21-day group. In 42- and 63-day groups, these cells were fewer than in 21-day group. Numerical difference was present between the groups, but statistical significance was not found between the groups. We concluded that nasal buserelin cytotoxicity was not potent in the nasal cavity in rabbits. We use nasal buserelin in all indications with confidence.