The adverse effects of cadmium on plants are accompanied by a limitation of photosynthesis, due to the production of reactive oxygen species, leading to oxidative damage to PSII and the disruption of key protein complexes involved in photosynthetic pathways. We investigated the effects of cadmium stress combined with high light in Arabidopsis thaliana, as dependent on the cadmium dose applied. The aim was to investigate the combined effect of the two stressors on photochemical processes with the hypothesis that Cd stress enhances the negative effect of the high light. The plants were treated with 0, 1, 10, and 50 mM Cd added as CdCl2 solution to soil (potted plants), and a high light stress. The highest dose (50 mM) induced a significant oxidative stress, reduced chlorophyll fluorescence parameters related to PSII functioning and increased energy dissipation mechanisms. Elevated Cd contents impaired the electron transport and limited PSII efficiency. OJIP analysis revealed a Cd-induced K- and L-band appearance documenting LHC-PSII limitation. The combination of Cd and high light stress resulted in the photoinhibition effects in PSII, i.e., a decrease in potential and effective yields of PSII.

• Keywords
• OJIP, cadmium, chlorophyll fluorescence, heavy metal, nonphotochemical quenching, photoinhibition, protective mechanisms,
• MeSH
• Arabidopsis * drug effects   radiation effects   physiology   metabolism   MeSH
• Chlorophyll metabolism   MeSH
• Photochemical Processes * drug effects   radiation effects   MeSH
• Photosynthesis * drug effects   radiation effects   MeSH
• Photosystem II Protein Complex metabolism   MeSH
• Stress, Physiological * drug effects   MeSH
• Cadmium * toxicity   pharmacology   MeSH
• Oxidative Stress drug effects   radiation effects   MeSH
• Reactive Oxygen Species metabolism   MeSH
• Light * MeSH
• Electron Transport drug effects   MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Chlorophyll MeSH
• Photosystem II Protein Complex MeSH
• Cadmium * MeSH
• Reactive Oxygen Species MeSH

In this study, we observed the effect of the application of soil dust enriched with risk elements (Cd, Pb, As and Zn) to leaf surfaces of lettuce (Lactuca sativa var. capitata) while it was grown under hydroponic conditions. This study aimed to determine how low soil dust particulate matter (PM) doses affected the activity of or damaged the photosynthetic apparatus and how the uptake of risk elements was associated with both epigenetic changes (5-methylcytosine content, i.e., 5mC) and stress metabolism. During the study, we obtained many results pertaining to risk element contents and biochemical (total phenolic content (TPC), malondialdehyde (MDA) content and the amount of free amino acids (AAs)) and physiological (photosynthesis parameters: net photosynthetic rate, transpiration rate, intercellular CO2 concentration, stomatal conductance, instantaneous water-use efficiency, maximum quantum yield of PSII, chlorophyll and carotenoid contents, and leaf water potential (WP)) plant features. The results showed an increase in MDA and 5mC. However, the transpiration rate, WP and free AAs decreased. In conclusion, contamination by very low doses of soil dust PM had no direct or significant effect on plant fitness, as shown by the TPC and 5mC content, which indicates that plants can overcome the oxidative stress caused by the accumulation of risk elements. From the above, we propose the use of epigenetic changes as biomarkers of potential changes in the activation of plant metabolism under stress caused by environmental pollution.

• MeSH
• Chlorophyll metabolism   MeSH
• Photosynthesis MeSH
• Plant Leaves metabolism   MeSH
• Particulate Matter * metabolism   MeSH
• Dust * MeSH
• Soil chemistry   MeSH
• Plants metabolism   MeSH
• Lactuca metabolism   MeSH
• Water metabolism   MeSH
• Environmental Pollution MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Chlorophyll MeSH
• Particulate Matter * MeSH
• Dust * MeSH
• Soil MeSH
• Water MeSH

Cadmium (Cd) is a major environmental contaminant due to its widespread industrial use. Cd contamination of soil and water is rather classical but has emerged as a recent problem. Cd toxicity causes a range of damages to plants ranging from germination to yield suppression. Plant physiological functions, i.e., water interactions, essential mineral uptake, and photosynthesis, are also harmed by Cd. Plants have also shown metabolic changes because of Cd exposure either as direct impact on enzymes or other metabolites, or because of its propensity to produce reactive oxygen species, which can induce oxidative stress. In recent years, there has been increased interest in the potential of plants with ability to accumulate or stabilize Cd compounds for bioremediation of Cd pollution. Here, we critically review the chemistry of Cd and its dynamics in soil and the rhizosphere, toxic effects on plant growth, and yield formation. To conserve the environment and resources, chemical/biological remediation processes for Cd and their efficacy have been summarized in this review. Modulation of plant growth regulators such as cytokinins, ethylene, gibberellins, auxins, abscisic acid, polyamines, jasmonic acid, brassinosteroids, and nitric oxide has been highlighted. Development of plant genotypes with restricted Cd uptake and reduced accumulation in edible portions by conventional and marker-assisted breeding are also presented. In this regard, use of molecular techniques including identification of QTLs, CRISPR/Cas9, and functional genomics to enhance the adverse impacts of Cd in plants may be quite helpful. The review's results should aid in the development of novel and suitable solutions for limiting Cd bioavailability and toxicity, as well as the long-term management of Cd-polluted soils, therefore reducing environmental and human health hazards.

• Keywords
• abiotic stress, cadmium, contamination, plant physiology and growth, remediation,
• Publication type
• Journal Article MeSH
• Review MeSH

BACKGROUND: Many metals are essential for plants and humans. Knowledge of metal distribution in plant tissues in vivo contributes to the understanding of physiological mechanisms of metal uptake, accumulation and sequestration. For those studies, X-rays are a non-destructive tool, especially suited to study metals in plants. RESULTS: We present microfluorescence imaging of trace elements in living plants using a customized benchtop X-ray fluorescence machine. The system was optimized by additional detector shielding to minimize stray counts, and by a custom-made measuring chamber to ensure sample integrity. Protocols of data recording and analysis were optimised to minimise artefacts. We show that Zn distribution maps of whole leaves in high resolution are easily attainable in the hyperaccumulator Noccaea caerulescens. The sensitivity of the method was further shown by analysis of micro- (Cu, Ni, Fe, Zn) and macronutrients (Ca, K) in non-hyperaccumulating crop plants (soybean roots and pepper leaves), which could be obtained in high resolution for scan areas of several millimetres. This allows to study trace metal distribution in shoots and roots with a wide overview of the object, and thus avoids making conclusions based on singular features of tiny spots. The custom-made measuring chamber with continuous humidity and air supply coupled to devices for imaging chlorophyll fluorescence kinetic measurements enabled direct correlation of element distribution with photosynthesis. Leaf samples remained vital even after 20 h of X-ray measurements. Subtle changes in some of photosynthetic parameters in response to the X-ray radiation are discussed. CONCLUSIONS: We show that using an optimized benchtop machine, with protocols for measurement and quantification tailored for plant analyses, trace metal distribution can be investigated in a reliable manner in intact, living plant leaves and roots. Zinc distribution maps showed higher accumulation in the tips and the veins of young leaves compared to the mesophyll tissue, while in the older leaves the distribution was more homogeneous.

• Keywords
• Arabidopsis halleri, Capsicum annuum, Glycine max, Hyperaccumulator, Leaf age, Micro X-ray fluorescence, Micronutrients, Noccaea caerulescens, Root, Zinc,
• Publication type
• Journal Article MeSH

Zinc is essential for the functioning of numerous proteins in plants. To investigate how Zn homeostasis interacts with virus infection, Zn-tolerant Noccaea ochroleucum plants exposed to deficient (Zn'0'), optimal (Zn10), and excess Zn (Zn100) concentrations, as well as Cd amendment, were infected with Turnip yellow mosaic virus (TYMV). Imaging analysis of fluorescence kinetics from the μs (OJIP) to the minutes (Kautsky effect, quenching analysis) time domain revealed strong patchiness of systemic virus-induced photosystem II (PSII) inhibition. That was more pronounced in Zn-deficient plants, while Zn excess acted synergistically with TYMV, in both cases resulting in reduced PSII reaction centers. Infected Cd-treated plants, already severely stressed, showed inhibited non-photochemical quenching and PSII activity. Quantitative in situ hybridization at the cellular level showed increased gene expression of ZNT5 and downregulation of HMA4 in infected Zn-deficient leaves. In Zn10 and Zn100 infected leaves, vacuolar sequestration of Zn increased by activation of HMA3 (mesophyll) and MTP1 (epidermis). This correlated with Zn accumulation in the mesophyll and formation of biomineralization dots in the cell wall (Zn100) visible by micro X-ray fluorescence tomography. The study reveals the importance of adequate Zn supply and distribution in the maintenance of photosynthesis under TYMV infection, achieved by tissue-targeted activation of metal transporter gene expression.

• Keywords
• TYMV infection, cadmium, chlorophyll fluorescence kinetics, metal transporters, micro X-ray fluorescence, non-hyperaccumulator, plant immunity, zinc,
• Publication type
• Journal Article MeSH

Solving the global environmental and agricultural problem of chronic low-level cadmium (Cd) exposure requires better mechanistic understanding. Here, soybean (Glycine max) plants were exposed to Cd concentrations ranging from 0.5 nM (background concentration, control) to 3 µM. Plants were cultivated hydroponically under non-nodulating conditions for 10 weeks. Toxicity symptoms, net photosynthetic oxygen production and photosynthesis biophysics (chlorophyll fluorescence: Kautsky and OJIP) were measured in young mature leaves. Cd binding to proteins [metalloproteomics by HPLC-inductively coupled plasma (ICP)-MS] and Cd ligands in light-harvesting complex II (LHCII) [X-ray absorption near edge structure (XANES)], and accumulation of elements, chloropyll, and metabolites were determined in leaves after harvest. A distinct threshold concentration of toxicity onset (140 nM) was apparent in strongly decreased growth, the switch-like pattern for nutrient uptake and metal accumulation, and photosynthetic fluorescence parameters such as Φ RE10 (OJIP) and saturation of the net photosynthetic oxygen release rate. XANES analyses of isolated LHCII revealed that Cd was bound to nitrogen or oxygen (and not sulfur) atoms. Nutrient deficiencies caused by inhibited uptake could be due to transporter blockage by Cd ions. The changes in specific fluorescence kinetic parameters indicate electrons not being transferred from PSII to PSI. Inhibition of photosynthesis combined with inhibition of root function could explain why amino acid and carbohydrate metabolism decreased in favour of molecules involved in Cd stress tolerance (e.g. antioxidative system and detoxifying ligands).

• Keywords
• Cadmium, XANES, lipidomics, metabolomics, metal stress, metalloproteomics, soybean (Glycine max), sublethal toxicity,
• MeSH
• Chlorophyll MeSH
• Photosynthesis MeSH
• Glycine max * MeSH
• Cadmium * toxicity   MeSH
• Plant Leaves MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Chlorophyll MeSH
• Cadmium * MeSH

Salicylic acid (SA) is a very simple phenolic compound (a C7H6O3 compound composed of an aromatic ring, one carboxylic and a hydroxyl group) and this simplicity contrasts with its high versatility and the involvement of SA in several plant processes either in optimal conditions or in plants facing environmental cues, including heavy metal (HM) stress. Nowadays, a huge body of evidence has unveiled that SA plays a pivotal role as plant growth regulator and influences intra- and inter-plant communication attributable to its methyl ester form, methyl salicylate, which is highly volatile. Under stress, including HM stress, SA interacts with other plant hormones (e.g., auxins, abscisic acid, gibberellin) and promotes the stimulation of antioxidant compounds and enzymes thereby alerting HM-treated plants and helping in counteracting HM stress. The present literature survey reviews recent literature concerning the roles of SA in plants suffering from HM stress with the aim of providing a comprehensive picture about SA and HM, in order to orientate the direction of future research on this topic.

• Keywords
• metal pollution, metal toxicity, ortho-hydroxybenzoic acid, plant hormone, polyphenols, signaling compound,
• MeSH
• Antioxidants metabolism   MeSH
• Stress, Physiological MeSH
• Salicylic Acid chemistry   metabolism   MeSH
• Metabolic Networks and Pathways MeSH
• Molecular Structure MeSH
• Oxidative Stress MeSH
• Reactive Oxygen Species metabolism   MeSH
• Plants drug effects   metabolism   MeSH
• Metals, Heavy adverse effects   chemistry   metabolism   MeSH
• Environmental Exposure * adverse effects   MeSH
• Publication type
• Journal Article MeSH
• Review MeSH
• Names of Substances
• Antioxidants MeSH
• Salicylic Acid MeSH
• Reactive Oxygen Species MeSH
• Metals, Heavy MeSH

Chlorophyll fluorescence kinetic analysis has become an important tool in basic and applied research on plant physiology and agronomy. While early systems recorded the integrated kinetics of a selected spot or plant, later systems enabled imaging of at least the slower parts of the kinetics (20-ms time resolution). For faster events, such as the rise from the basic dark-adapted fluorescence yield to the maximum (OJIP transient), or the fluorescence yield decrease during reoxidation of plastoquinone A after a saturating flash, integrative systems are used because of limiting speed of the available imaging systems. In our new macroscopic and microscopic systems, the OJIP or plastonique A reoxidation fluorescence transients are directly imaged using an ultrafast camera. The advantage of such systems compared to nonimaging measurements is the analysis of heterogeneity of measured parameters, for example between the photosynthetic tissue near the veins and the tissue further away from the veins. Further, in contrast to the pump-and-probe measurement, direct imaging allows for measuring the transition of the plant from the dark-acclimated to a light-acclimated state via a quenching analysis protocol in which every supersaturating flash is coupled to a measurement of the fast fluorescence rise. We show that pump-and-probe measurement of OJIP is prone to artifacts, which are eliminated with the direct measurement. The examples of applications shown here, zinc deficiency and cadmium toxicity, demonstrate that this novel imaging platform can be used for detection and analysis of a range of alterations of the electron flow around PSII.

• MeSH
• Arabidopsis cytology   metabolism   MeSH
• Brassicaceae cytology   drug effects   metabolism   MeSH
• Chlorophyll chemistry   metabolism   MeSH
• Equipment Design MeSH
• Fluorescence MeSH
• Microscopy, Fluorescence instrumentation   methods   MeSH
• Photosynthesis MeSH
• Glycine max cytology   drug effects   metabolism   MeSH
• Kinetics MeSH
• Plant Leaves cytology   MeSH
• Mesophyll Cells metabolism   MeSH
• Plastoquinone metabolism   MeSH
• Zinc metabolism   MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Chlorophyll MeSH
• Plastoquinone MeSH
• Zinc MeSH

P1B-ATPases are decisive for metal accumulation phenotypes, but mechanisms of their regulation are only partially understood. Here, we studied the Cd/Zn transporting ATPases NcHMA3 and NcHMA4 from Noccaea caerulescens as well as AhHMA3 and AhHMA4 from Arabidopsis halleri. Protein biochemistry was analyzed on HMA4 purified from roots of N. caerulescens in active state. Metal titration of NcHMA4 protein with an electrochromic dye as charge indicator suggested that HMA4 reaches maximal ATPase activity when all internal high-affinity Cd2+ binding sites are occupied. Although HMA4 was reported to be mainly responsible for xylem loading of heavy metals for root to shoot transport, the current study revealed high expression of NcHMA4 in shoots as well. Further, there were additional 20 and 40 kD fragments at replete Zn2+ and toxic Cd2+, but not at deficient Zn2+ concentrations. Altogether, the protein level expression analysis suggested a more multifunctional role of NcHMA4 than previously assumed. Organ-level transcription analysis through quantitative PCR of mRNA in N. caerulescens and A. halleri confirmed the strong shoot expression of both NcHMA4 and AhHMA4. Further, in shoots NcHMA4 was more abundant in 10 μM Zn2+ and AhHMA4 in Zn2+ deficiency. In roots, NcHMA4 was up-regulated in response to deficient Zn2+ when compared to replete Zn2+ and toxic Cd2+ treatment. In both species, HMA3 was much more expressed in shoots than in roots, and HMA3 transcript levels remained rather constant regardless of Zn2+ supply, but were up-regulated by 10 μM Cd2+. Analysis of cellular expression by quantitative mRNA in situ hybridisation showed that in A. halleri, both HMA3 and HMA4 mRNA levels were highest in the mesophyll, while in N. caerulescens they were highest in the bundle sheath of the vein. This is likely related to the different final storage sites for hyperaccumulated metals in both species: epidermis in N. caerulescens, mesophyll in A. halleri.

• Keywords
• QISH, RT-qPCR, cadmium, heavy metal ATPase, metal hyperaccumulator plants, zinc,
• Publication type
• Journal Article MeSH

Changes of photosynthetic activity in vivo of individual heterocysts and vegetative cells in the diazotrophic cyanobacterium Anabaena sp. strain PCC 7120 during the course of diazotrophic acclimation were determined using fluorescence kinetic microscopy (FKM). Distinct phases of stress and acclimation following nitrogen step-down were observed. The first was a period of perception, in which the cells used their internally stored nitrogen without detectable loss of PS II activity or pigments. In the second, the stress phase of nitrogen limitation, the cell differentiation occurred and an abrupt decline of fluorescence yield was observed. This decline in fluorescence was not paralleled by a corresponding decline in photosynthetic pigment content and PS II activity. Both maximal quantum yield and sustained electron flow were not altered in vegetative cells, only in the forming heterocysts. The third, acclimation phase started first in the differentiating heterocysts with a recovery of PS II photochemical yields [Formula: see text] Afterwards, the onset of nitrogenase activity was observed, followed by the restoration of antenna pigments in the vegetative cells, but not in the heterocysts. Surprisingly, mature heterocysts were found to have an intact PS II as judged by photochemical yields, but a strongly reduced PS II-associated antenna as judged by decreased F 0. The possible importance of the functional PS II in heterocysts is discussed. Also, the FKM approach allowed to follow in vivo and evaluate the heterogeneity in photosynthetic performance among individual vegetative cells as well as heterocysts in the course of diazotrophic acclimation. Some cells along the filament (so-called "superbright cells") were observed to display transiently increased fluorescence yield, which apparently proceeded by apoptosis.

• MeSH
• Acclimatization physiology   MeSH
• Anabaena cytology   physiology   MeSH
• Single-Cell Analysis methods   MeSH
• Pigments, Biological metabolism   MeSH
• Chlorophyll metabolism   MeSH
• Nitrogen deficiency   MeSH
• Microscopy, Fluorescence methods   MeSH
• Photosynthesis physiology   MeSH
• Stress, Physiological MeSH
• Kinetics MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Names of Substances
• Pigments, Biological MeSH
• Chlorophyll MeSH
• Nitrogen MeSH