Abinaya Manivannan

Blue LED light enhances growth, phytochemical contents, and antioxidant enzyme activities of Rehmannia glutinosa cultured in vitro

The objective of the current study is to determine the effect of light quality on enhancement of growth, phytochemicals, antioxidant potential, and antioxidant enzyme activities at in vitro cultures of Rehmannia glutinosa Libosch. In vitro-grown shoot tip explants were cultured on the plant growth regulator (PGR)-free Murashige and Skoog (MS) medium and cultured under a conventional cool white fluorescent light (control), blue light emitting diode (LED) light or red LED light. After four weeks, the growth traits along with total phenol content, total flavonoid content, free radical scavenging activities, and antioxidant enzyme activities were measured. Interestingly, the blue or red LED treatments showed a significant increase in growth parameters compared with the cool white florescent light. In addition, the LED treatments increased the total phenol and flavonoid levels in leaf and root extracts. Furthermore, data on the total antioxidant capacity, reducing power potential, and DPPH radical scavenging capacity also revealed the enhancement of antioxidant capacity under both blue and red LED treatments. Especially, the blue LED treatment significantly increased the antioxidant enzyme activities in both the leaf and root, followed by the red LED treatment. Modulation in the spectral quality particularly by the blue LED induced the antioxidant defense line and was directly correlated with the enhancement of phytochemicals. Therefore, the incorporation of blue or red LED light sources during in vitro propagation of R. glutinosa can be a beneficial way to increase the medicinal values of the plant.

Physiological and Proteomic Analysis in Chloroplasts of Solanum lycopersicum L. under Silicon Efficiency and Salinity Stress

Tomato plants often grow in saline environments in Mediterranean countries where salt accumulation in the soil is a major abiotic stress that limits its productivity. However, silicon (Si) supplementation has been reported to improve tolerance against several forms of abiotic stress. The primary aim of our study was to investigate, using comparative physiological and proteomic approaches, salinity stress in chloroplasts of tomato under silicon supplementation. Tomato seedlings (Solanum lycopersicum L.) were grown in nutrient media in the presence or absence of NaCl and supplemented with silicon for 5 days. Salinity stress caused oxidative damage, followed by a decrease in silicon concentrations in the leaves of the tomato plants. However, supplementation with silicon had an overall protective effect against this stress. The major physiological parameters measured in our studies including total chlorophyll and carotenoid content were largely decreased under salinity stress, but were recovered in the presence of silicon. Insufficient levels of net-photosynthesis, transpiration and stomatal conductance were also largely improved by silicon supplementation. Proteomics analysis of chloroplasts analyzed by 2D-BN-PAGE (second-dimensional blue native polyacrylamide-gel electrophoresis) revealed a high sensitivity of multiprotein complex proteins (MCPs) such as photosystems I (PSI) and II (PSII) to the presence of saline. A significant reduction in cytochrome b6/f and the ATP-synthase complex was also alleviated by silicon during salinity stress, while the complex forms of light harvesting complex trimers and monomers (LHCs) were rapidly up-regulated. Our results suggest that silicon plays an important role in moderating damage to chloroplasts and their metabolism in saline environments. We therefore hypothesize that tomato plants have a greater capacity for tolerating saline stress through the improvement of photosynthetic metabolism and chloroplast proteome expression after silicon supplementation.

Silicon alleviates salt stress by modulating antioxidant enzyme activities in Dianthus caryophyllus ‘Tula’

Although silicon (Si) is not considered as an essential element, it is beneficial to the plant growth. Its effect is more evident under abiotic and biotic stress conditions. The objective of this study is to investigate the role of Si on the in vitro growth and resistance to salt stress of Dianthus caryophyllus ‘Tula’. The experiment was designed as a factorial design with 0, 50, or 100 mg·L−1 of potassium silicate (K2SiO3) in combination with 0, 50, or 100 mM sodium chloride (NaCl). The treatment of 50 mg·L−1 Si improved the growth of plant. However, the treatment of Si at 100 mg·L−1 reduced the growth. Although NaCl retarded the growth, addition of Si along with NaCl to the culture medium mitigated the effect of NaCl. A primary defense line by Si to overcome the photosynthetic depression was apparent from the increased chlorophyll content in the Si + NaCl treatment as compared to the treatment of NaCl alone. Enhancement of growth and resistance to salinity by Si was thought to be due to the modulation in activity of antioxidant enzymes, such as superoxide dismutase, ascorbate peroxidase, guaiacol peroxidase, and catalase. Therefore, our results suggested that 50 mg·L−1 Si supplementation could be optimal for improved growth in vitro and enhanced resistance against salinity in D. caryophyllus ‘Tula’.

Silicon Mitigates Salinity Stress by Regulating the Physiology, Antioxidant Enzyme Activities, and Protein Expression in Capsicum annuum ‘Bugwang’

Silicon- (Si-) induced salinity stress resistance was demonstrated at physiological and proteomic levels in Capsicum annuum for the first time. Seedlings of C. annuum were hydroponically treated with NaCl (50 mM) with or without Si (1.8 mM) for 15 days. The results illustrated that saline conditions significantly reduced plant growth and biomass and photosynthetic parameters and increased the electrolyte leakage potential, lipid peroxidation, and hydrogen peroxide level. However, supplementation of Si allowed the plants to recover from salinity stress by improving their physiology and photosynthesis. During salinity stress, Si prevented oxidative damage by increasing the activities of antioxidant enzymes. Furthermore, Si supplementation recovered the nutrient imbalance that had occurred during salinity stress. Additionally, proteomic analysis by two-dimensional gel electrophoresis (2DE) followed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) revealed that Si treatment upregulated the accumulation of proteins involved in several metabolic processes, particularly those associated with nucleotide binding and transferase activity. Moreover, Si modulated the expression of vital proteins involved in ubiquitin-mediated nucleosome pathway and carbohydrate metabolism. Overall, the results illustrate that Si application induced resistance against salinity stress in C. annuum by regulating the physiology, antioxidant metabolism, and protein expression.

Exogenous Supplementation of Silicon Improved the Recovery of Hyperhydric Shoots in Dianthus caryophyllus L. by Stabilizing the Physiology and Protein Expression

Hyperhydricity is one of the major problems hindering in vitro propagation of Dianthus caryophyllus L. Silicon (Si) is a well-known beneficial element renowned for its stress amelioration properties in plants. This study has demonstrated the physiological and molecular mechanism behind the Si-mediated recovery from hyperhydricity in D. caryophyllus L. ‘Green Beauty’. Four weeks old hyperhydric shoots obtained from temporary immersion system were cultured on the Murashige and Skoog medium supplemented with 0 (control), 1.8 mM, or 3.6 mM of potassium silicate (K2SiO3). After 2 weeks of culture, we observed only 20% of hyperhydric shoots were recovered in control. On the other hand hyperhydricity, shoot recovery percentage in 1.8 mM and 3.6 mM of Si were 44% and 36%, respectively. Shoots in control possessed higher lipid peroxidation rate compared to the Si treatments. Similarly, damaged stomata were detected in the control, while Si treatments restored the normal stomatal development. Expressions of superoxide dismutase, guaiacol peroxidase, and catalase varied between the control and Si treatments. Furthermore, a proteomic analysis showed that as compared with the control Si up-regulated 17 and 10 protein spots in abundance at 1.8 and 3.6 mM of Si, respectively. In comparison to the 3.6 mM, 1.8 mM of Si treatment up-regulated 19 proteins and down-regulated 7 proteins. Identified proteins were categorized into six groups according to their biological roles such as ribosomal binding, oxido-reduction, hormone/cell signaling, metal/ion binding, defense, and photosynthesis. The proteomic results revealed that Si actively involved in the various metabolisms to accelerate the recovery of the shoots from hyperhydricity. Thus, the outcomes of this study can be utilized for addressing the molecular insight of hyperhydricity and its recovery mechanism by the supplementation of Si. Therefore, we conclude that active involvement of Si in the regulation and signaling process of proteins at 1.8 mM concentration could be efficient to trigger the reclamation process of hyperhydric carnation shoots.

Binding Mode Investigation of Polyphenols from Scrophularia Targeting Human Aldose Reductase Using Molecular Docking and Molecular Dynamics Simulations

Aldose reductase (ALR2), a vital enzyme involved in polyol pathway, has befitted as a novel drug target in antidiabetes drug discovery process. In the present study, the binding mode and pharmacokinetic properties of potential polyphenolic compounds with reported aldose reductase inhibitory activity from the genus Scrophularia have been investigated. The human ALR2 enzyme (PDB ID: 2FZD) acted as the receptor in the current study. Among the compounds investigated, acacetin, a methoxy flavonoid, displayed the stable binding to the active site of ALR2 with least binding energy value. Molecular interaction analysis revealed that acacetin interrupts the proton donation mechanism, necessary for the catalytic activity of ALR2, by forming H-bond with Tyr48 (proton donor). In addition, acacetin also possessed favorable ADME properties and complies with Lipinski’s rule of 5 representing the possible drug-like nature compared to other polyphenols. Interestingly, the biological activity predictions also ranked acacetin with higher probability score for aldose reductase inhibition activity. Moreover, the molecular dynamics simulation of ALR2-acacetin complex was validated for the stability of ligand binding and the refined complex was used for generation of receptor-ligand pharmacophore model. Thus, the molecular insights of receptor-ligand interactions gained from the present study can be utilized for the development of novel aldose reductase inhibitors from Scrophularia.

Chemical Elicitor-Induced Modulation of Antioxidant Metabolism and Enhancement of Secondary Metabolite Accumulation in Cell Suspension Cultures of Scrophularia kakudensis Franch

Scrophularia kakudensis is an important medicinal plant with pharmaceutically valuable secondary metabolites. To develop a sustainable source of naturaceuticals with vital therapeutic importance, a cell suspension culture was established in S. kakudensis for the first time. Friable calli were induced from the leaf explants cultured on a Murashige and Skoog (MS) medium containing 3.0 mg·L−1 6-benzyladenine (BA) in a combination with 2 mg·L−1 2,4-dichlorophenoxy acetic acid (2,4-D). From the callus cultures, a cell suspension culture was initiated and the cellular differentiation was investigated. In addition, the effect of biotic elicitors such as methyl jasmonate (MeJa), salicylic acid (SA), and sodium nitroprusside (SNP) on the accumulation of secondary metabolites and antioxidant properties was demonstrated. Among the elicitors, the MeJa elicited the accumulation of total phenols, flavonoids, and acacetin, a flavonoid compound with multiple pharmaceutical values. Similarly, the higher concentrations of the MeJa significantly modulated the activities of antioxidant enzymes and enhanced the scavenging potentials of free radicals of cell suspension extracts. Overall, the outcomes of this study can be utilized for the large scale production of pharmaceutically important secondary metabolites from S. kakudensis through cell suspension cultures.

Silicon-mediated enhancement of physiological and biochemical characteristics of Zinnia elegans ‘Dreamland Yellow’ grown under salinity stress

This study investigated the effects of silicon (Si) nutrition on hydroponically grown Zinnia elegans under salinity stress. In this study, six treatments, the control (basal nutrients without NaCl or Si), Si 50 (1.8 mM), Si 100 (3.6 mM), NaCl 50 (50 mM), Si 50 + NaCl 50 (1.8 mM Si; 50 mM NaCl), and Si 100 + NaCl 50 (Si-3.6 mM + NaCl-50 mM), were employed. After 15 days of treatment, growth parameters, biochemical measurements, and antioxidant enzyme activities were examined. Salinity stress significantly reduced plant growth, biomass, photosynthetic parameters, and pigments, and increased the electrolyte leakage potential (ELP), lipid peroxidation, and hydrogen peroxide level. Interestingly, with Si supplementation, Z. elegans recovered from salinity stress. Si enhanced growth and photosynthesis, and prevented the decomposition of photosynthetic pigments. Moreover, the addition of Si increased membrane integrity, thereby reducing the ELP and lipid peroxidation levels under salinity stress. Furthermore, Si modulated the activity of the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), and guaiacol peroxidase (GPX) in scavenging excess reactive oxygen species (ROS). Additionally, Si increased the macronutrient and micronutrient contents. Therefore, augmentation with Si provided salinity resistance and enhanced the growth of Z. elegans.

Blue and red light-emitting diodes improve the growth and physiology of in vitro-grown carnations ‘Green Beauty’ and ‘Purple Beauty’

The objective of this study was to determine the effect of light quality on the growth, physiology, and antioxidant enzyme activity of two important domestic carnation (Dianthus caryophyllus) cultivars: ‘Green Beauty’ and ‘Purple Beauty’. In vitro-grown shoot tip explants were cultured on the plant growth regulator (PGR)-free Murashige and Skoog (MS) medium under a conventional cool white fluorescent lamp (control), blue light-emitting diode (LED), or red LED. Growth traits, photosynthetic and biochemical parameters, activity of antioxidant enzymes, and nutrient content were measured after 8 weeks. Interestingly, the blue and red LED treatments resulted in a significant increase in growth, photosynthetic parameters, and nutrient content in comparison to the conventional cool white florescent lamp treatment. In addition, red LED treatment increased the activities of antioxidant enzymes and elemental contents in both cultivars. Thus, incorporating blue or red LED treatments enhances the quality of ‘Green Beauty’ and ‘Purple Beauty’ carnations propagated in vitro.

In Vitro Propagation, Phytochemical Analysis, and Evaluation of Free Radical Scavenging Property of Scrophularia kakudensis Franch Tissue Extracts

The current study deals with in vitro propagation, antioxidant property estimation, and assessment of acacetin content in Scrophularia kakudensis Franch. Adventitious shoot induction was achieved from the nodal explant with the highest number of adventitious shoots per explant (17.4) on Murashige and Skoogs (MS) medium fortified with 2.0 mg·L−1 6-benzyladenine (BA) and 0.5 mg L−1 indole-3-acetic acid (IAA). Maximum number of roots per plant (16.5) was noted in half strength MS medium supplemented with 0.5 mg·L−1 IAA. The regenerated plants displayed successful survival ratio (95%) in the greenhouse. The highest content of acacetin, a pharmaceutically important flavonoid, was observed in the shoot extracts (in vitro: 32.83 µg·g−1 FW; in vivo: 30.05 µg·g−1 FW) followed by root extracts. Total phenol and flavonoid contents along with free radical scavenging assays revealed the occurrence of larger amount of antioxidants in shoot extract in comparison with callus and root extracts of S. kakudensis. Thus, the outcome of the present study can be highly beneficial for the germplasm conservation and commercial cultivation of S. kakudensis for therapeutic purposes.