Comparing cadmium-induced effects on the regulation of the DNA damage response and cell cycle progression between entire rosettes and individual leaves of Arabidopsis thaliana.

Cadmium (Cd) activates the DNA damage response (DDR) and inhibits the cell cycle in Arabidopsis thaliana through the transcription factor SUPPRESSOR OF GAMMA RESPONSE 1. The aim of this study was to investigate which individual leaf best reflects the Cd-induced effects on the regulation of the DDR and cell cycle progression in rosettes, enabling a more profound interpretation of the rosette data since detailed information, provided by the individual leaf responses, is lost when studying the whole rosette. Wild-type A. thaliana plants were cultivated in hydroponics and exposed to different Cd concentrations. Studied individual leaves were leaf 1 and 2, which emerged before Cd exposure, and leaf 3, which emerged upon Cd exposure. The DDR and cell cycle regulation were studied in rosettes as well as individual leaves after several days of Cd exposure. Varying concentration-dependent response patterns were observed between the entire rosette and individual leaves. Gene expression of selected DDR and cell cycle regulators showed higher similarity in their response between the rosette and the individual leaf emerged during Cd exposure than between both individual leaves. The same pattern was observed for plant growth and cell cycle-related parameters. We conclude that Cd-induced effects on the regulation of the DDR and cell cycle progression in the leaf that emerged during Cd exposure, resemble those observed in the rosette the most, which contributes to the interpretation of the rosette data in the framework of plant development and after exposure to Cd.

The glutathione-dependent alarm triggers signalling responses involved in plant acclimation to cadmium.

Cadmium (Cd) uptake from polluted soils inhibits plant growth and disturbs physiological processes, at least partly due to disturbances in the cellular redox environment. Although the sulfur-containing antioxidant glutathione is important in maintaining redox homeostasis, its role as an antioxidant can be overruled by its involvement in Cd chelation as a phytochelatin precursor. Following Cd exposure, plants rapidly invest in phytochelatin production, thereby disturbing the redox environment by transiently depleting glutathione concentrations. Consequently, a network of signalling responses is initiated, in which the phytohormone ethylene is an important player involved in the recovery of glutathione levels. Furthermore, these responses are intricately connected to organellar stress signalling and autophagy, and contribute to cell fate determination. In general, this may pave the way for acclimation (e.g. restoration of glutathione levels and organellar homeostasis) and plant tolerance in the case of mild stress conditions. This review addresses connections between these players and discusses the possible involvement of the gasotransmitter hydrogen sulfide in plant acclimation to Cd exposure.

Cadmium-induced oxidative stress responses and acclimation in plants require fine-tuning of redox biology at subcellular level.

Cadmium (Cd) is one of the most toxic compounds released into our environment and is harmful to human health, urging the need to remediate Cd-polluted soils. To this end, it is important to increase our insight into the molecular mechanisms underlying Cd stress responses in plants, ultimately leading to acclimation, and to develop novel strategies for economic validation of these soils. Albeit its non-redox-active nature, Cd causes a cellular oxidative challenge, which is a crucial determinant in the onset of diverse signalling cascades required for long-term acclimation and survival of Cd-exposed plants. Although it is well known that Cd affects reactive oxygen species (ROS) production and scavenging, the contribution of individual organelles to Cd-induced oxidative stress responses is less well studied. Here, we provide an overview of the current information on Cd-induced organellar responses with special attention to redox biology. We propose that an integration of organellar ROS signals with other signalling pathways is essential to finetune plant acclimation to Cd stress.

Redox-mediated responses to high temperature in plants.

As sessile organisms, plants are particularly affected by climate change and will face more frequent and extreme temperature variations in the future. Plants have developed a diverse range of mechanisms allowing them to perceive and respond to these environmental constraints, which requires sophisticated signalling mechanisms. Reactive oxygen species (ROS) are generated in plants exposed to various stress conditions including high temperatures and are presumed to be involved in stress response reactions. The diversity of ROS-generating pathways and the ability of ROS to propagate from cell to cell and to diffuse through cellular compartments and even across membranes between subcellular compartments put them at the centre of signalling pathways. In addition, their capacity to modify the cellular redox status and to modulate functions of target proteins, notably through cysteine oxidation, show their involvement in major stress response transduction pathways. ROS scavenging and thiol reductase systems also participate in the transmission of oxidation-dependent stress signals. In this review, we summarize current knowledge on the functions of ROS and oxidoreductase systems in integrating high temperature signals, towards the activation of stress responses and developmental acclimation mechanisms.

Short-term effects of cadmium on leaf growth and nutrient transport in rice plants.

Consumption of rice grains contaminated with high concentrations of cadmium (Cd) can cause serious long-term health problems. Moreover, even low Cd concentrations present in the soil can result in the abatement of plant performance, leading to lower grain yield. Studies examining the molecular basis of plant defense against Cd-induced oxidative stress could pave the way in creating superior rice varieties that display an optimal antioxidative defense system to cope with Cd toxicity. In this study, we showed that after one day of Cd exposure, hydroponically grown rice plants exhibited adverse shoot biomass and leaf growth effects. Cadmium accumulates especially in the roots and the leaf meristematic region, leading to a disturbance of manganese homeostasis in both the roots and leaves. The leaf growth zone showed an increased amount of lipid peroxidation indicating that Cd exposure disturbed the oxidative balance. We propose that an increased expression of genes related to the glutathione metabolism such as glutathione synthetase 2, glutathione reductase and phytochelatin synthase 2, rather than genes encoding for antioxidant enzymes, is important in combating early Cd toxicity within the leaves of rice plants. Furthermore, the upregulation of two RESPIRATORY BURST OXIDASE HOMOLOG genes together with a Cd concentration-dependent increase of abscisic acid might cause stomatal closure or cell wall modification, potentially leading to the observed leaf growth reduction. Whereas abscisic acid was also elevated at long term exposure, a decrease of the growth hormone auxin might further contribute to growth inhibition and concomitantly, an increase in salicylic acid might stimulate the activity of antioxidative enzymes after a longer period of Cd exposure. In conclusion, a clear interplay between phytohormones and the oxidative challenge affect plant growth and acclimation during exposure to Cd stress.

Efficient regulation of copper homeostasis underlies accession-specific sensitivities to excess copper and cadmium in roots of Arabidopsis thaliana.

The commonly used Arabidopsis thaliana natural accessions Columbia (Col-0) and Wassilewskija (Ws) are known to differ in their metal sensitivity, with Col-0 being more sensitive to copper (Cu) and cadmium (Cd) than Ws. As both Cu and Cd are known to affect Cu homeostasis, it was investigated whether this process is part of an accession-specific mechanism underlying their difference in metal sensitivity. As roots are the first contact point during metal exposure, responses were compared between roots of both accessions of hydroponically grown plants exposed to excess Cu or Cd for 24 and 72 h. Root Cu levels increased in both accessions under Cu and Cd exposure. However, under Cu exposure, the downregulation of Cu transporter (COPT) genes in combination with a more pronounced upregulation of metallothionein gene MT2b indicated that Ws plants coped better with the elevated Cu concentrations. The Cd-induced disturbance in Cu homeostasis was more efficiently counteracted in roots of Ws plants than in Col-0 plants. This was indicated by a higher upregulation of the SPL7-mediated pathway, crucial in the regulation of the Cu homeostasis response. In conclusion, maintaining the Cu homeostasis response in roots is key to accession-specific differences in Cu and Cd sensitivity.

At the Crossroads of Survival and Death: The Reactive Oxygen Species-Ethylene-Sugar Triad and the Unfolded Protein Response.

Upon stress, a trade-off between plant growth and defense responses defines the capacity for survival. Stress can result in accumulation of misfolded proteins in the endoplasmic reticulum (ER) and other organelles. To cope with these proteotoxic effects, plants rely on the unfolded protein response (UPR). The involvement of reactive oxygen species (ROS), ethylene (ETH), and sugars, as well as their crosstalk, in general stress responses is well established, yet their role in UPR deserves further scrutiny. Here, a synopsis of current evidence for ROS-ETH-sugar crosstalk in UPR is discussed. We propose that this triad acts as a major signaling hub at the crossroads of survival and death, integrating information from ER, chloroplasts, and mitochondria, thereby facilitating a coordinated stress response.

Glutathione Is Required for the Early Alert Response and Subsequent Acclimation in Cadmium-Exposed Arabidopsis thaliana Plants.

Pollution by cadmium (Cd) is a worldwide problem, posing risks to human health and impacting crop yield and quality. Cadmium-induced phytotoxicity arises from an imbalance between antioxidants and pro-oxidants in favour of the latter. The Cd-induced depletion of the major antioxidant glutathione (GSH) strongly contributes to this imbalance. Rather than being merely an adverse effect of Cd exposure, the rapid depletion of root GSH levels was proposed to serve as an alert response. This alarm phase is crucial for an optimal stress response, which defines acclimation later on. To obtain a better understanding on the importance of GSH in the course of these responses and how these are defined by the rapid GSH depletion, analyses were performed in the GSH-deficient cadmium-sensitive 2-1 (cad2-1) mutant. Cadmium-induced root and leaf responses related to oxidative challenge, hydrogen peroxide (H2O2), GSH, ethylene, and 1-aminocyclopropane-1-carboxylic acid (ACC) were compared between wild-type (WT) and mutant Arabidopsis thaliana plants. Although the cad2-1 mutant has significantly lower GSH levels, root GSH depletion still occurred, suggesting that the chelating capacity of GSH is prioritised over its antioxidative function. We demonstrated that responses related to GSH metabolism and ACC production were accelerated in mutant roots and that stress persisted due to suboptimal acclimation. In general, the redox imbalance in cad2-1 mutant plants and the lack of proper transient ethylene signalling contributed to this suboptimal acclimation, resulting in a more pronounced Cd effect.

Long-Term Cd Exposure Alters the Metabolite Profile in Stem Tissue of Medicago sativa.

As a common pollutant, cadmium (Cd) is one of the most toxic heavy metals accumulating in agricultural soils through anthropogenic activities. The uptake of Cd by plants is the main entry route into the human food chain, whilst in plants it elicits oxidative stress by unbalancing the cellular redox status. Medicago sativa was subjected to chronic Cd stress for five months. Targeted and untargeted metabolic analyses were performed. Long-term Cd exposure altered the amino acid composition with levels of asparagine, histidine and proline decreasing in stems but increasing in leaves. This suggests tissue-specific metabolic stress responses, which are often not considered in environmental studies focused on leaves. In stem tissue, profiles of secondary metabolites were clearly separated between control and Cd-exposed plants. Fifty-one secondary metabolites were identified that changed significantly upon Cd exposure, of which the majority are (iso)flavonoid conjugates. Cadmium exposure stimulated the phenylpropanoid pathway that led to the accumulation of secondary metabolites in stems rather than cell wall lignification. Those metabolites are antioxidants mitigating oxidative stress and preventing cellular damage. By an adequate adjustment of its metabolic composition, M. sativa reaches a new steady state, which enables the plant to acclimate under chronic Cd stress.

Identifying the Pressure Points of Acute Cadmium Stress Prior to Acclimation in Arabidopsis thaliana.

The toxic metal cadmium (Cd) is a major soil pollutant. Knowledge on the acute Cd-induced stress response is required to better understand the triggers and sequence of events that precede plant acclimation. Therefore, we aimed to identify the pressure points of Cd stress using a short-term exposure set-up ranging from 0 h to 24 h. Acute responses related to glutathione (GSH), hydrogen peroxide (H2O2), 1-aminocyclopropane-1-carboxylic acid (ACC), ethylene and the oxidative challenge were studied at metabolite and/or transcript level in roots and leaves of Arabidopsis thaliana either exposed or not to 5 µM Cd. Cadmium rapidly induced root GSH depletion, which might serve as an alert response and modulator of H2O2 signalling. Concomitantly, a stimulation of root ACC levels was observed. Leaf responses were delayed and did not involve GSH depletion. After 24 h, a defined oxidative challenge became apparent, which was most pronounced in the leaves and concerted with a strong induction of leaf ACC synthesis. We suggest that root GSH depletion is required for a proper alert response rather than being a merely adverse effect. Furthermore, we propose that roots serve as command centre via a.o. root-derived ACC/ethylene to engage the leaves in a proper stress response.

Cadmium inhibits cell cycle progression and specifically accumulates in the maize leaf meristem.

It is well known that cadmium (Cd) pollution inhibits plant growth, but how this metal impacts leaf growth processes at the cellular and molecular level is still largely unknown. In the current study, we show that Cd specifically accumulates in the meristematic tissue of the growing maize leaf, while Cd concentration in the elongation zone rapidly declines as the deposition rates diminish and cell volumes increase due to cell expansion. A kinematic analysis shows that, at the cellular level, a lower number of meristematic cells together with a significantly longer cell cycle duration explain the inhibition of leaf growth by Cd. Flow cytometry analysis suggests an inhibition of the G1/S transition, resulting in a lower proportion of cells in the S phase and reduced endoreduplication in expanding cells under Cd stress. Lower cell cycle activity is also reflected by lower expression levels of key cell cycle genes (putative wee1, cyclin-B2-4, and minichromosome maintenance4). Cell elongation rates are also inhibited by Cd, which is possibly linked to the inhibited endoreduplication. Taken together, our results complement studies on Cd-induced growth inhibition in roots and link inhibited cell cycle progression to Cd deposition in the leaf meristem.

Glutathione: A key player in metal chelation, nutrient homeostasis, cell cycle regulation and the DNA damage response in cadmium-exposed Arabidopsis thaliana.

Glutathione (GSH) is an important player in plant responses to cadmium (Cd) through its dual function as an antioxidant and precursor for metal-chelating phytochelatins (PCs). In addition, it was shown to be involved in cell cycle regulation in Arabidopsis thaliana roots, but its involvement in this process in leaves is largely unknown and has never been evaluated in Cd-exposed plants. This study aimed to elucidate the role of GSH in leaf growth and development, metal chelation, nutrient homeostasis and cell cycle regulation in A. thaliana plants upon prolonged Cd exposure. Responses were compared between wild-type (WT) plants and three GSH-deficient mutants. Our results indicate that PC production remains important in plants exposed to Cd for an extended duration. Furthermore, an important role for GSH in regulating nutrient homeostasis in Cd-exposed plants was revealed. Cell cycle analysis demonstrated that negative effects of Cd exposure on cell division and endoreplication were more pronounced in leaves of the GSH-deficient cadmium-sensitive 2-1 (cad2-1) mutant in comparison to the WT, indicating the involvement of GSH in cell cycle regulation. Finally, a crucial role for GSH in transcriptional activation of the Cd-induced DNA damage response (DDR) was revealed, as the Cd-induced upregulation of DDR-related genes was either less pronounced or completely abolished in leaves of the GSH-deficient mutants.

Suppressor of Gamma Response 1 Modulates the DNA Damage Response and Oxidative Stress Response in Leaves of Cadmium-Exposed Arabidopsis thaliana.

Cadmium (Cd) exposure causes an oxidative challenge and inhibits cell cycle progression, ultimately impacting plant growth. Stress-induced effects on the cell cycle are often a consequence of activation of the DNA damage response (DDR). The main aim of this study was to investigate the role of the transcription factor SUPPRESSOR OF GAMMA RESPONSE 1 (SOG1) and three downstream cyclin-dependent kinase inhibitors of the SIAMESE-RELATED (SMR) family in the Cd-induced DDR and oxidative challenge in leaves of Arabidopsis thaliana. Effects of Cd on plant growth, cell cycle regulation and the expression of DDR genes were highly similar between the wildtype and smr4/5/7 mutant. In contrast, sog1-7 mutant leaves displayed a much lower Cd sensitivity within the experimental time-frame and significantly less pronounced upregulations of DDR-related genes, indicating the involvement of SOG1 in the Cd-induced DDR. Cadmium-induced responses related to the oxidative challenge were disturbed in the sog1-7 mutant, as indicated by delayed Cd-induced increases of hydrogen peroxide and glutathione concentrations and lower upregulations of oxidative stress-related genes. In conclusion, our results attribute a novel role to SOG1 in regulating the oxidative stress response and connect oxidative stress to the DDR in Cd-exposed plants.

Assessment of the Antioxidative Potential of Rosmarinus officinalis L. (Lamiaceae) Irrigated with Static Magnetic Field-Treated Water

Abstract Phenolic compounds are one of the main groups of secondary metabolites in plants and are known for their antioxidant activity. Rosmarinus officinalis L. (rosemary) contains different phenolic compounds including carnosol, carnosic acid and rosmarinic acid. In Cuba, rosemary cultivation is limited because it is difficult to propagate and has a low yield. As a result, it was removed from the Herbal Medicine National Formulary. However, the National Public Health System has a strong interest in rosemary because of its value as a natural antioxidant medicine. Irrigation with water treated with a static magnetic field (SMF) is a possible strategy to increase rosemary yield. This technology has been applied to accelerate plant growth and increase crop quality. The aim of this study was to evaluate the content of phenolic compounds and antioxidant activity in aqueous leaf extracts from plants irrigated with SMF-treated water in comparison to control plants. Significant differences in phenolic content and antioxidant activity were observed between aqueous extracts of control plants and plants irrigated with SMF-treated water. Therefore, irrigation with SMF-treated water is a promising technology to improve the cultivation of rosemary as a raw material to obtain pharmaceutical products with high antioxidant activities.

Cadmium and Plant Development: An Agony from Seed to Seed.

Anthropogenic pollution of agricultural soils with cadmium (Cd) should receive adequate attention as Cd accumulation in crops endangers human health. When Cd is present in the soil, plants are exposed to it throughout their entire life cycle. As it is a non-essential element, no specific Cd uptake mechanisms are present. Therefore, Cd enters the plant through transporters for essential elements and consequently disturbs plant growth and development. In this review, we will focus on the effects of Cd on the most important events of a plant's life cycle covering seed germination, the vegetative phase and the reproduction phase. Within the vegetative phase, the disturbance of the cell cycle by Cd is highlighted with special emphasis on endoreduplication, DNA damage and its relation to cell death. Furthermore, we will discuss the cell wall as an important structure in retaining Cd and the ability of plants to actively modify the cell wall to increase Cd tolerance. As Cd is known to affect concentrations of reactive oxygen species (ROS) and phytohormones, special emphasis is put on the involvement of these compounds in plant developmental processes. Lastly, possible future research areas are put forward and a general conclusion is drawn, revealing that Cd is agonizing for all stages of plant development.