Melatonin and strigolactone mitigate chromium toxicity through modulation of ascorbate-glutathione pathway and gene expression in tomato.

Chromium (Cr) is considered one of the most hazardous metal contaminant reducing crop production and putting human health at risk. Phytohormones are known to regulate chromium stress, however, the function of melatonin and strigolactones in Chromium stress tolerance in tomato is rarely investigated. Here we investigated the potential role of melatonin (ML) and strigolactone (SL) on mitigating Chromium toxicity in tomato. With exposure to 300 µM Cr stress a remarkable decline in growth (63.01%), biomass yield (50.25)%, Pigment content (24.32%), photosynthesis, gas exchange and Physico-biochemical attributes of tomato was observed. Cr treatment also resulted in oxidative stress closely associated with higher H2O2 generation (215.66%), Lipid peroxidation (50.29%), electrolyte leakage (440.01%) and accumulation of osmolytes like proline and glycine betine. Moreover, Cr toxicity up-regulated the transcriptional expression profiles of antioxidant, stress related and metal transporter genes and down-regulated the genes related to photosynthesis. The application of ML and SL alleviated the Cr induced phytotoxic effects on photosynthetic pigments, gas exchange parameters and restored growth of tomato plants. ML and SL supplementation induced plant defense system via enhanced regulation of antioxidant enzymes, ascorbate and glutathione pool and transcriptional regulation of several genes. The coordinated regulation of antioxidant and glyoxalase systems expressively suppressed the oxidative stress. Hence, ML and SL application might be considered as an effective approach for minimizing Cr uptake and its detrimental effects in tomato plants grown in contaminated soils. The study may also provide new insights into the role of transcriptional regulation in the protection against heavy metal toxicity.

Mitigation of Negative Effects of Chromium (VI) Toxicity in Faba Bean (Vicia faba) Plants through the Supplementation of Kinetin (KN) and Gibberellic Acid (GA3).

The present study was carried out to explore the possible role of kinetin and gibberellic acid (GA3) on faba bean under chromium (Cr) stress. Cr treatment negatively affected growth and biomass production, reduced photosynthetic pigments, and inhibited photosynthesis, gas exchange parameters, antioxidant enzymes, and the glyoxylase cycle. Moreover, Cr stress enhanced the production of malondialdehyde (MDA, 216.11%) and hydrogen peroxide (H2O2, 230.16%), electrolyte leakage (EL, 293.30%), and the accumulation of proline and glycine betaine. Exogenous application of kinetin and GA3 increased growth and biomass, improved pigment contents and photosynthesis, as well as up-regulated the antioxidant system by improving the antioxidant enzyme activities and the content of nonenzymatic components, and the glyoxylase cycle. Additionally, kinetin and GA3 application displayed a considerable enhancement in proline (602.61%) and glycine betaine (423.72), which help the plants to maintain water balance under stress. Furthermore, a decline in Cr uptake was also observed due to kinetin and GA3 application. Exogenous application of kinetin and GA3 ameliorated the toxic effects of Cr in faba bean plants, up-shooting the tolerance mechanisms, including osmolyte metabolism and the antioxidant system.

Co-over expression of Ascorbate Glutathione pathway enzymes improve mercury tolerance in tomato.

The genetic modification of plants for the removal of inorganic pollutants from contaminated soil and water bodies is an emerging area for addressing environmental concerns. This approach is based on the ability of plants to take up and accumulate heavy metals, with efficiency being dependent on the underlying mechanisms of heavy metal accumulation and tolerance. A robust antioxidant pathway is determinantal for heavy metal uptake and accumulation and, therefore, in this study, we evaluated the transgenic tomato plants installed with Ascorbate Glutathione (ASA-GSH) pathway genes for uptake, accumulation, and response to mercury (Hg). We observed that ASA-GSH overexpressing lines were resilient to Hg stress as they displayed higher photosynthetic activity and increased photosynthetic gas exchange parameters with a concomitant decrease in ion leakage under Hg stress. Additionally, transgenic lines accumulated high osmolytes and showed enhanced activity of antioxidant enzymes. Moreover, the results of SEM and confocal microscopy confirmed least damage to plant tissue in ASA-GSH overexpressing lines compared to wild-type under Hg-stress which was further supported by Atomic absorption study that revealed a significant decline in Hg accumulation in the leaves of transgenic lines compared to wild-type under stress conditions. In conclusion, pyramiding of ASA-GSH pathway genes in tomato plants is an efficient approach for the development of Hg-resistant tomato plants and the reclamation of Hg-contaminated sites.

Ascorbate glutathione antioxidant system alleviates fly ash stress by modulating growth physiology and biochemical responses in Solanum lycopersicum.

Tomato plants (Solanum lycopersicum L.) were developed in soils with different fly ash (FA) amendments (25, 50, 75, 100% FA) to measure the effects of FA on metal accumulation, chlorophyll pigments, chlorophyll fluorescence, growth, biomass, gas exchange parameters, and the ascorbate glutathione pathway (AsA-GSH). The metal concentration was much higher in FA compared to the garden soil/(control). The observed metal translocation was higher in roots than shoots. Plants raised in soils treated with 50% or more FA showed significant decreases in growth, biomass, gas exchange parameters, protein, chlorophyll pigments, and fluorescence parameters. Additionally, a significant increase in antioxidants under higher FA-amended soils were observed. Our results showed that the ability of Solanum lycopersicum plants to effectively synchronize the actions of antioxidant enzymes associated in reactive oxygen species (ROS) scavenging - notably superoxidase dismutase (SOD), ascorbate peroxidase (APX), and glutathione reductase (GR) - with good maintenance of the AsA/DHA ratio, that could be connected to FA stress tolerance. The toxic metals present in FA caused oxidative stress in Solanum lycopersicum, as evident from the increase in electrolyte leakage (EL), lipid peroxidation (MDA), and ROS levels. Furthermore, the AsA-GSH cycle plays a key role in alleviating oxidative damage caused by FA application.

Pyramiding ascorbate-glutathione pathway in Lycopersicum esculentum confers tolerance to drought and salinity stress.

KEY MESSAGE: Stacking Glutathione-Ascorbate pathway genes (PgSOD, PgAPX, PgGR, PgDHAR and PgMDHAR) under stress inducible promoter RD29A imparts significant tolerance to drought and salinity stress in Solanum lycopersicum. Although the exposure of plants to different environmental stresses results in overproduction of reactive oxygen species (ROS), many plants have developed some unique systems to alleviate the ROS production and mitigate its deleterious effect. One of the key pathways that gets activated in plants is ascorbate glutathione (AsA-GSH) pathway. To demonstrate the effect of this pathway in tomato, we developed the AsA-GSH overexpression lines by stacking the genes of the AsA-GSH pathway genes isolated from Pennisetum glaucoma (Pg) including PgSOD, PgAPX, PgGR, PgDHAR and PgMDHAR under stress inducible promoter RD29A. The overexpression lines have an improved germination and seedling growth with concomitant elevation in the survival rate. The exposure of transgenic seedlings to varying stress regiments exhibited escalation in the antioxidant enzyme activity and lesser membrane damage as reflected by decreased electrolytic leakage and little accumulation of malondialdehyde and H2O2. Furthermore, the transgenic lines accumulated high levels of osmoprotectants with increase in the relative water content. The increased photosynthetic activity and enhanced gaseous exchange parameters further confirmed the enhanced tolerance of AsA-GSH overexpression lines. We concluded that pyramiding of AsA-GSH pathway genes is an effective strategy for developing stress resistant crops.

Jasmonic acid (JA) and gibberellic acid (GA3) mitigated Cd-toxicity in chickpea plants through restricted cd uptake and oxidative stress management.

Cadmium stress is one of the chief environmental cues that can substantially reduce plant growth. In the present research, we studied the effect of jasmonic acid (JA) and gibberellic acid (GA3) applied individually and/or in combination to chickpea (Cicer arietinum) plants exposed to 150 µM cadmium sulphate. Cadmium stress resulted in reduced plant growth and pigment contents. Moreover, chickpea plants under cadmium contamination displayed higher levels of electrolytic leakage, H2O2, and malonaldehyde, as well as lower relative water content. Plants primed with JA (1 nM) and those foliar-fed with GA3 (10-6 M) showed improved metal tolerance by reducing the accumulation of reactive oxygen species, malonaldehyde and electrolytic leakage, and increasing relative water content. . Osmoprotectants like proline and glycinebetaine increased under cadmium contamination. Additionally, the enzymatic activities and non-enzymatic antioxidant levels increased markedly under Cd stress, but application of JA as well as of GA3 further improved these attributes. Enzymes pertaining to the ascorbate glutathione and glyoxylase systems increased significantly when the chickpea plants were exposed to Cd. However, JA and GA3 applied singly or in combination showed improved enzymatic activities as well as nutrient uptake, whereas they reduced the metal accumulation in chickpea plants. Taken together, our findings demonstrated that JA and GA3 are suitable agents for regulating Cd stress resistance in chickpea plants.

Ectopic expression of a novel cold-resistance protein 1 from Brassica oleracea promotes tolerance to chilling stress in transgenic tomato.

Cold stress is considered as one of the major environmental factors that adversely affects the plant growth and distribution. Therefore, there arises an immediate need to cultivate effective strategies aimed at developing stress-tolerant crops that would boost the production and minimise the risks associated with cold stress. In this study, a novel cold-responsive protein1 (BoCRP1) isolated from Brassica oleracea was ectopically expressed in a cold susceptible tomato genotype Shalimar 1 and its function was investigated in response to chilling stress. BoCRP1 was constitutively expressed in all the tissues of B. oleracea including leaf, root and stem. However, its expression was found to be significantly increased in response to cold stress. Moreover, transgenic tomato plants expressing BoCRP1 exhibited increased tolerance to chilling stress (4 °C) with an overall improved rate of seed germination, increased root length, reduced membrane damage and increased accumulation of osmoprotectants. Furthermore, we observed increased transcript levels of stress responsive genes and enhanced accumulation of reactive oxygen species scavenging enzymes in transgenic plants on exposure to chilling stress. Taken together, these results strongly suggest that BoCRP1 is a promising candidate gene to improve the cold stress tolerance in tomato.

Impact of drought and heat stress individually and in combination on physio-biochemical parameters, antioxidant responses, and gene expression in Solanum lycopersicum.

The present study was carried out to investigate the effect of individual drought, heat, and combined drought and heat stress on tomato plants. Combined stress resulted in the higher accumulation of Proline (101.9%), MDA (38.55%), H2O2 (101.19%), and lower levels of RWC (53.84%). Individual drought and heat stress decreased photosynthetic pigments like total chlorophyll content by (45.45%) and (25.35%), respectively, higher rates of pigment reduction (79.42%) were observed under combined drought and heat stress. Combined stress decreased PSII efficiency (Fv/Fm), quantum yield (ΦPSII), and photochemical efficiency (qp) and increased non-photochemical quenching (NPQ) levels. Moreover, the gas exchange parameters E, A, and Pn decreased by 5.36%, 36.45%, and 51.00%, respectively, in comparison to control plants. Antioxidant enzymes, SOD, APX, CAT, and GR showed a two- to threefold increase under combined drought and heat stress; however, the non-enzymatic antioxidants AsA and GSH displayed one-twofold increase under combined stress. Moreover, 2- to 2.5-fold decrease was observed in MDHAR and DHAR enzyme transcripts under combined stress conditions. The transcripts corresponding to AsA-GSH pathway enzymes SOD, APX, GR, DHAR, and MDHAR were up-regulated by 8- to 12-fold under combined drought and heat. Furthermore, DREB and LEA transcripts were up-regulated under drought and combined stress and down-regulated under drought stress. In the same manner, HSP70 and HSP90 transcripts were up-regulated under heat and combined stress; however, the transcription levels got down-regulated under drought stress. Additionally, rbcL and RCA transcripts were down-regulated especially under combined stress in comparison to individual drought and heat conditions. PSIP680 relative expression levels were up-regulated under drought stress; however, the transcripts were down-regulated under heat and combined stress. Taken together, the results suggest that the combined stress has a predominant effect over individual stress.

Fly-Ash Pollution Modulates Growth, Biochemical Attributes, Antioxidant Activity and Gene Expression in Pithecellobium Dulce (Roxb) Benth.

This study investigates the effect of fly ash (FA) on the Pithecellobium dulce (Roxb) Benth. trees growing at three different locations. FA stress caused significant changes in different leaf attributes like sugar, protein contents, photosynthetic pigments, nitrate content and nitrate reductase activity in foliar tissues of plants growing at a highly contaminated site, as compared to a low-pollution site. Lower rates of stomatal conductance (SC) were observed in P. dulce leaves under fly ash stress conditions that drastically reduced net photosynthetic rate (PN); however, intercellular carbon dioxide concentration and stomatal index (SI) showed an increase under the same stress conditions. On the other hand, significant increase was also observed in the proline, sulphur and nitrogen contents. A significant increase in oxidative stress and, consequently, in antioxidant enzymes such as ascorbate peroxidase (APX), catalase (CAT), peroxidase (POD), and superoxidase dismutase (SOD) and Air pollution tolerance index were discovered at three different sites. The transcriptional expression of antioxidant and stress responsive genes was higher at HPS as compared to two other two sites of the study. Taken together the results demonstrated that the P. dulce is best suited as a fly ash stress tolerant plant species with the potential to provide an alternative for the reclamation of fly ash affected soils.

24-Epibrassinolide (EBR) Confers Tolerance against NaCl Stress in Soybean Plants by Up-Regulating Antioxidant System, Ascorbate-Glutathione Cycle, and Glyoxalase System.

: The present research was performed to assess the effect of 24-epibrassinolide (EBR) on salt-stressed soybean plants. Salt stress suppressed growth, biomass yield, gas exchange parameters, pigment content, and chlorophyll fluorescence, but all these parameters were up-regulated by EBR supply. Moreover, salt stress increased hydrogen peroxide, malondialdehyde, and electrolyte leakage. EBR supplementation reduced the accumulation of oxidative stress biomarkers. The activities of superoxide dismutase and catalase, and the accumulation of proline, glycinebetaine, total phenols, and total flavonoids increased with NaCl stress, but these attributes further increased with EBR supplementation. The activities of enzymes and the levels of non-enzymatic antioxidants involved in the Asc-Glu cycle also increased with NaCl stress, and further enhancement in these attributes was recorded by EBR supplementation. Salinity elevated the methylglyoxal content, but it was decreased by the EBR supplementation accompanying with up-regulation of the glyoxalase cycle (GlyI and GlyII). Salinity enhanced the Na+ uptake in root and shoot coupled with a decrease in uptake of Ca2+, K+, and P. However, EBR supplementation declined Na+ accumulation and promoted the uptake of the aforementioned nutrients. Overall, EBR supplementation regulated the salt tolerance mechanism in soybean plants by modulating osmolytes, activities of key enzymes, and the levels of non-enzymatic antioxidants.

Green Ultrasound versus Conventional Synthesis and Characterization of Specific Task Pyridinium Ionic Liquid Hydrazones Tethering Fluorinated Counter Anions: Novel Inhibitors of Fungal Ergosterol Biosynthesis.

A series of specific task ionic liquids (ILs) based on a pyridiniumhydrazone scaffold in combination with hexafluorophosphate (PF6-), tetrafluoroboron (BF4-) and/or trifluoroacetate (CF3COO-) counter anion, were designed and characterized by IR, NMR and mass spectrometry. The reactions were conducted under both conventional and green ultrasound procedures. The antifungal potential of the synthesized compounds 2-25 was investigated against 40 strains of Candida (four standard and 36 clinical isolates). Minimum inhibitory concentrations (MIC90) of the synthesized compounds were in the range of 62.5-2000 µg/mL for both standard and oral Candida isolates. MIC90 results showed that the synthesized 1-(2-(4-chlorophenyl)-2-oxoethyl)-4-(2-(4-fluorobenzylidene)hydrazinecarbonyl)-pyridin-1-ium hexafluorophosphate (11) was found to be most effective, followed by 4-(2-(4-fluorobenzylidene)hydrazinecarbonyl)-1-(2-(4-nitrophenyl)-2-oxoethyl)-pyridin-1-ium hexafluorophosphate (14) and 1-(2-ethoxy-2-oxoethyl)-4-(2-(4-fluorobenzylidene)hydrazinecarbonyl)pyridin-1-ium hexafluorophosphate (8). All the Candida isolates showed marked sensitivity towards the synthesized compounds. Ergosterol content was drastically reduced by more active synthesized compounds, and agreed well with MIC90 values. Confocal scanning laser microscopy (CLSM) results showed that the red colored fluorescent dye enters the test agent treated cells, which confirms cell wall and cell membrane damage. The microscopy results obtained suggested membrane-located targets for the action of these synthesized compounds. It appears that the test compounds might be interacting with ergosterol in the fungal cell membranes, decreasing the membrane ergosterol content and ultimately leading to membrane disruption as visible in confocal results. The present study indicates that these synthesized compounds show significant antifungal activity against Candida which forms the basis to carry out further in vivo experiments before their clinical use.

Cinnamaldehyde and its derivatives, a novel class of antifungal agents.

The last few decades have seen an alarming rise in fungal infections, which currently represent a global health threat. Despite extensive research towards the development of new antifungal agents, only a limited number of antifungal drugs are available in the market. The routinely used polyene agents and many azole antifungals are associated with some common side effects such as severe hepatotoxicity and nephrotoxicity. Also, antifungal resistance continues to grow and evolve and complicate patient management, despite the introduction of new antifungal agents. This suitation requires continuous attention. Cinnamaldehyde has been reported to inhibit bacteria, yeasts, and filamentous molds via the inhibition of ATPases, cell wall biosynthesis, and alteration of membrane structure and integrity. In this regard, several novel cinnamaldehyde derivatives were synthesized with the claim of potential antifungal activities. The present article describes antifungal properties of cinnamaldehyde and its derivatives against diverse classes of pathogenic fungi. This review will provide an overview of what is currently known about the primary mode of action of cinnamaldehyde. Synergistic approaches for boosting the effectiveness of cinnamaldehyde and its derivatives have been highlighted. Also, a keen analysis of the pharmacologically active systems derived from cinnamaldehyde has been discussed. Finally, efforts were made to outline the future perspectives of cinnamaldehyde-based antifungal agents. The purpose of this review is to provide an overview of current knowledge about the antifungal properties and antifungal mode of action of cinnamaldehyde and its derivatives and to identify research avenues that can facilitate implementation of cinnamaldehyde as a natural antifungal.

Morphological and biochemical changes in Azadirachta indica from coal combustion fly ash dumping site from a thermal power plant in Delhi, India.

The foliar and biochemical traits of Azadirachta indica A. Juss from fly ash (FA) dumping site in Badarpur thermal power plant (BTPP) New Delhi, India was studied. Three different experimental sites were selected at different distances from the thermal power plant. Ambient suspended particulate matter (SPM) and plant responses such as leaf pigments (chlorophyll a, chlorophyll b, and carotenoids), total chlorophyll, net photosynthetic rate, stomatal index (SI), stomatal conductance (SC), intercellular carbon dioxide concentration [CO2]i, net photosynthetic rate (NPR), nitrogen, nitrate, nitrate reductase activity, proline, protein, reducing sugar and sulphur content were measured. Considerable reduction in pigments (chlorophyll a, chlorophyll b and carotenoids), and total chlorophyll was observed at fly ash dumping site. Fly ash stress revealed the inhibitory effect on Nitrate reductase activity (NRA), Nitrate, soluble protein, and reducing sugar content, whereas stimulatory effect was found for the stomatal index, nitrogen, proline, antioxidants and sulphur content in the leaves. Under fly ash stress, stomatal conductance was low, leading to declining in photosynthetic rate and increase in the internal CO2 concentration of leaf. Single leaf area (SLA), leaf length and leaf width also showed a declining trend from control to the polluted site. Antioxidant enzymes increased in leaves reflecting stress and extenuation of reactive oxygen species (ROS).

Impaired ergosterol biosynthesis mediated fungicidal activity of Co(II) complex with ligand derived from cinnamaldehyde.

In this study, we have used aldehyde function of cinnamaldehyde to synthesize N, N'-Bis (cinnamaldehyde) ethylenediimine [C20H20N2] and Co(II) complex of the type [Co(C40H40N4)Cl2]. The structures of the synthesized compounds were determined on the basis of physiochemical analysis and spectroscopic data ((1)H NMR, FTIR, UV-visible and mass spectra) along with molar conductivity measurements. Anticandidal activity of cinnamaldehyde its ligand [L] and Co(II) complex was investigated by determining MIC80, time-kill kinetics, disc diffusion assay and ergosterol extraction and estimation assay. Ligand [L] and Co(II) complex are found to be 4.55 and 21.0 folds more efficient than cinnamaldehyde in a liquid medium. MIC80 of Co(II) complex correlated well with ergosterol inhibition suggesting ergosterol biosynthesis to be the primary site of action. In comparison to fluconazole, the test compounds showed limited toxicity against H9c2 rat cardiac myoblasts. In confocal microscopy propidium iodide (PI) penetrates the yeast cells when treated with MIC of metal complex, indicating a disruption of cell membrane that results in imbibition of dye. TEM analysis of metal complex treated cells exhibited notable alterations or damage to the cell membrane and the cell wall. The structural disorganization within the cell cytoplasm was noted. It was concluded that fungicidal activity of Co(II) complex originated from loss of membrane integrity and a decrease in ergosterol content is only one consequence of this.