Toxic effect of mercury on arbuscular mycorrhizal fungi colonisation and physiological status of three seed-based Miscanthus hybrids.

BACKGROUND: Currently, mercury pollution is a widespread problem in the world. As mercury is difficult to remove from the environment, it has long-term negative effects on soil health and human life. One of the techniques to stabilise Hg is phytostabilisation, which can be supported by arbuscular mycorrhizal fungi (AMF). METHODS: In a 4-month pot experiment, we investigated the suitability of three seed-based Miscanthus hybrids (GNT3, GNT34, GNT43) for growth on soils heavily polluted with mercury (6795.7 mg kg-1). During the experiment, the effects of high soil contamination with mercury on physiological parameters and colonisation of roots of seed-based Miscanthus hybrids by indigenous AMF from Hg-contaminated and uncontaminated soils were investigated. RESULTS: A high pseudo-total Hg concentration (6795.75 mg kg-1) in soil was found. The Hg content in the aerial part of GNT34 grown on Hg-contaminated soil was 1.5 times and 3 times higher than GNT3 and GNT43, respectively. The Hg content in the roots of GNT3 on Hg-contaminated soil was 25% and 10% lower than that of GNT34 and GNT43, respectively. The N content in the aboveground part of GNT34 in the Hg variant was 13.5% lower compared to the control soil. The P and K content in the shoots of the Miscanthus hybrids was lower in the plants grown on Hg-contaminated soil. The P content in GNT43 in the Hg variant was 33% and 19% lower than in GNT34 and GNT3, respectively. The K content in GNT34 in the Hg variant was 24.7% and 31.4% higher than in GNT43 and GNT3, respectively. The dry weight of the shoots and roots as well as the shoot height of the Miscanthus hybrids were lower in Hg-contaminated soil. Lower values of AMF root colonisation parameters (F, M) were observed in the plants in the Hg variant. In the Hg variant, a lower photosynthetic rate and a decrease in chlorophyll content were observed in the leaves of the Miscanthus hybrids. In the Hg variant, an increase in the content of flavonols was observed. The strongest toxic effect of mercury on the light phase of photosynthesis was measured in GNT34. CONCLUSION: Soils heavily contaminated with mercury negatively affected the physiological parameters of Miscanthus, as evidenced by a decrease in photosynthetic rate and biomass. The ability of indigenous AMF from Hg-contaminated soils to colonise the roots of seed-based Miscanthus hybrids was limited.

Heat shock protein 90 (Hsp90) inhibitor STA-9090 (Ganetespib) ameliorates inflammation in a mouse model of atopic dermatitis.

Molecular chaperones belonging to the heat shock protein 90 (Hsp90) family are implicated in inflammatory processes and described as potential novel therapeutic targets in autoimmune/inflammatory skin diseases. While the pathological role of circulating Hsp90 has been recently proposed in patients with atopic dermatitis (AD), a chronic inflammatory skin disease characterized by intense itching and recurrent skin lesions, studies aimed at investigating the role of Hsp90 as a potential target of AD therapy have not yet been conducted. Here, the effects of the Hsp90 blocker STA-9090 (Ganetespib) applied systemically or topically were determined in an experimental mouse model of dinitrochlorobenzene (DNCB)-induced AD. Intraperitoneal administration of STA-9090 ameliorated clinical disease severity, histological epidermal thickness, and dermal leukocyte infiltration in AD mice which was associated with reducing the scratching behavior in DNCB-treated animals. Additionally, topically applied STA-9090 led to lowered disease activity in AD mice, reduced serum levels of IgE, and up-regulated filaggrin expression in lesional skin samples. Our observations suggest that Hsp90 may be a promising therapeutic target in atopic dermatitis and potentially other inflammatory or autoimmune dermatoses.

The Effect of Combined Drought and Temperature Stress on the Physiological Status of Calcareous Grassland Species as Potential Candidates for Urban Green Infrastructure.

Nature-based solutions are promising for climate adaptation and environmental management in urban areas, but urban conditions are stressful for vegetation. In particular, the interaction of drought and high temperatures may be detrimental. Guiding plant selection for urban greening with native species requires a far better knowledge of plant adaptations and stress acclimation. We tested the physiological responses of four candidate calcareous grassland species for green roofs and walls to the combined effects of drought and high temperatures under controlled conditions. The tested species proved relatively resistant to stress despite different strategies to protect the photosynthetic apparatus, maintain water balance, and repair damages. Based on the physiological responses, we rank the species in descending order of resistance to the stress factors tested: Trifolium medium > Festuca ovina > Carex flacca > Potentilla reptans, but all four can serve as potential candidates for green walls and roofs. Physiological stress screening of plant species for use on green roofs and walls supplements the habitat template approach to provide a stronger and wider base for prioritizations.

The effect of combined drought and trace metal elements stress on the physiological response of three Miscanthus hybrids.

Drought is a serious threat worldwide and has a significant impact on agricultural production and soil health. It can pose an even greater threat when it involves land contaminated with trace metal element (TMEs). To prevent desertification, such land should be properly managed and growing Miscanthus for energy or raw material purposes could be a solution. The effects of drought and TMEs were studied in a pot experiment on three different Miscanthus hybrids (conventional Miscanthus × giganteus, TV1 and GNT10) considering growth parameters, photosynthetic parameters and elemental composition of roots, rhizomes and shoots. GNT10 was characterised by the weakest gas exchange among the hybrids, which was compensated by the highest number of leaves and biomass. The strongest correlations between the studied parameters were found for TV1, which might indicate a high sensitivity to TME stress. For M × g and GNT10, the main mechanisms for coping with stress seem to be biomass management through number of shoots and leaves and gas exchange. The main factor determining the extent of accumulation of TMEs was the amount of water applied in the experimental treatment, which was related to the location of the plant in the aniso-isohydric continuum. GNT10 was the most resistant plant to combined stress, while it responded similarly to TV1 when drought and trace metal elements were applied separately.

Differences in the detection of circulating Hsp90 alpha between patients with atopic dermatitis and dermatitis herpetiformis.

Heat shock protein 90 alpha (Hsp90α) is one of the key intra- and extracellular chaperones responsible for the biological activity of various signaling molecules that are involved in (auto)immune-mediated inflammatory diseases. Recent epidemiologic data suggest that patients with atopic dermatitis (AD) are at risk for several autoimmune diseases, including dermatitis herpetiformis (DH), an extraintestinal manifestation of celiac disease (CD). In addition, pruritic diseases such as AD may be confused clinically with DH. In this study, we aimed to determine the role of circulating Hsp90α in patients with AD in relation to patients with DH, CD, and healthy controls. Using an enzyme-linked immunosorbent assay, levels of circulating Hsp90α were determined in serum samples derived from patients with AD (n = 31), DH (n = 26), CD (n = 15), and healthy controls (n = 55). Although serum concentrations of Hsp90α were similar between patients with DH, CD, and healthy controls, we found that serum levels of Hsp90α were significantly higher (mean value of 5.08-fold; p < 0.0001) in patients with AD when compared to patients with DH. A cutoff value calculated as 2 × standard deviation above the mean concentration of Hsp90α in DH patients revealed that 83.9% (26/31) of AD patients were Hsp90α positive, whereas none of the DH patients (0/26) displayed such a positivity. This preliminary study suggests a distinct role for extracellular Hsp90α in the pathogenesis of AD compared to DH and its potential use in distinguishing AD from DH. Nevertheless, the potential role of the evaluation of extracellular Hsp90α for distinguishing between AD and DH is at present speculative and requires further and careful observations.

Altered Gene Expression of IL-35 and IL-36α in the Skin of Patients with Atopic Dermatitis.

A comprehensive understanding of atopic dermatitis (AD) pathogenesis is desired, especially in the current era of novel biologics and small molecule drugs. In recent years, new cytokines have emerged that may play a significant role in the pathogenesis of AD. Using the tape stripping (TS) method, this study analyzed the gene expression of IL-35 and IL-36α in lesional and nonlesional AD skin compared with healthy skin and their association with the clinical features of AD among the Polish population. Ten AD patients and seven healthy individuals were enrolled. The lesional skin of the AD patients showed significantly higher expression levels of IL-35 compared to healthy skin (p = 0.0001). The expression level of IL-36α was significantly higher in lesional AD skin than in nonlesional AD skin (p = 0.0039) and healthy skin (p = 0.0045). There was a significant negative correlation between AD severity and the expression level of IL-35 in both lesional (R = -0.66, p = 0.048) and nonlesional skin (R = -0.9, p = 0.0016). In summary, both IL-35 and IL-36α appear to play a role in the pathogenesis of AD. Furthermore, it might be speculated that IL-35 and IL-36α may be potential candidates for disease biomarkers. However, further studies are needed to verify these assumptions and comprehensively elucidate their importance in the pathogenesis of AD.

Heat Shock Protein 90 (Hsp90) and Hsp70 as Potential Therapeutic Targets in Autoimmune Skin Diseases.

Over a hundred different autoimmune diseases have been described to date, which can affect every organ in the body, including the largest one, the skin. In fact, up to one-fifth of the world's population suffers from chronic, noninfectious inflammatory skin diseases, the development of which is significantly influenced by an autoimmune response. One of the hallmarks of autoimmune diseases is the loss of immune tolerance, which leads to the formation of autoreactive lymphocytes or autoantibodies and, consequently, to chronic inflammation and tissue damage. The treatment of autoimmune skin diseases mainly focuses on immunosuppression (using, e.g., corticosteroids) but almost never leads to the development of permanent mechanisms of immune tolerance. In addition, current therapies and their long-term administration may cause serious adverse effects. Hence, safer and more effective therapies that bring sustained balance between pro- and anti-inflammatory responses are still desired. Both intra- and extracellular heat shock proteins (Hsps), specifically well-characterized inducible Hsp90 and Hsp70 chaperones, have been highlighted as therapeutic targets for autoimmune diseases. This review presents preclinical data on the involvement of Hsp90 and Hsp70 in modulating the immune response, specifically in the context of the treatment of selected autoimmune skin diseases with emphasis on autoimmune bullous skin diseases and psoriasis.

Biological aspects of phage therapy versus antibiotics against Salmonella enterica serovar Typhimurium infection of chickens.

Phage therapy is a promising alternative treatment of bacterial infections in human and animals. Nevertheless, despite the appearance of many bacterial strains resistant to antibiotics, these drugs still remain important therapeutics used in human and veterinary medicine. Although experimental phage therapy of infections caused by Salmonella enterica was described previously by many groups, those studies focused solely on effects caused by bacteriophages. Here, we compared the use of phage therapy (employing a cocktail composed of two previously isolated and characterized bacteriophages, vB_SenM-2 and vB_Sen-TO17) and antibiotics (enrofloxacin and colistin) in chickens infected experimentally with S. enterica serovar Typhimurium. We found that the efficacies of both types of therapies (i.e. the use of antibiotics and phage cocktail) were high and very similar to one another when the treatment was applied shortly (one day) after the infection. Under these conditions, S. Typhimurium was quickly eliminated from the gastrointestinal tract (GIT), to the amount not detectable by the used methods. However, later treatment (2 or 4 days after detection of S. Typhimurium in chicken feces) with the phage cocktail was significantly less effective. Bacteriophages remained in the GIT for up to 2-3 weeks, and then were absent in feces and cloaca swabs. Interestingly, both phages could be found in various organs of chickens though with a relatively low abundance. No development of resistance of S. Typhimurium to phages or antibiotics was detected during the experiment. Importantly, although antibiotics significantly changed the GIT microbiome of chickens in a long-term manner, analogous changes caused by phages were transient, and the microbiome normalized a few weeks after the treatment. In conclusion, phage therapy against S. Typhimurium infection in chickens appeared as effective as antibiotic therapy (with either enrofloxacin or colistin), and less invasive than the use the antibiotics as fewer changes in the microbiome were observed.

Field Evaluation of Arbuscular Mycorrhizal Fungal Colonization in Miscanthus × giganteus and Seed-Based Miscanthus Hybrids Grown in Heavy-Metal-Polluted Areas.

Understanding the behavior of arbuscular mycorrhizal fungi (AMF) associated with plants is essential for optimizing plant cultivation to the phytoremediation of degraded soils. The objective of the present study was to evaluate the differences in AMF root colonization between novel seed-based interspecific Miscanthus hybrids (M. sacchariflorus × M. sinensis) and the standard M. × giganteus when grown in soils contaminated with heavy metals (Pb, Cd, and Zn). During the third and fourth growing seasons, higher concentration of metals in the roots and a limited transfer of metals from the roots to the shoots were observed in all the plants studied. After the third growing season, the lowest values of AMF colonization rates were observed for the GNT34 hybrid. After the fourth growing season, AMF colonization decreased, which could be due to the drought that occurred during that season. GNT34 showed a lower tendency to develop mycorrhizal structures on heavy-metal (HM)-contaminated soils than GNT41 and M × g; however, this hybrid was insensitive to changes in colonization rates during the dry growing season.

Pathological Relevance of Anti-Hsp70 IgG Autoantibodies in Epidermolysis Bullosa Acquisita.

Stress-induced heat shock protein 70 (Hsp70) is a key intra- and extracellular molecular chaperone implicated in autoimmune processes. Highly immunogenic extracellular Hsp70 can activate innate and acquired (adaptive) immune responses driving the generation of anti-Hsp70 autoantibodies that are frequently observed in inflammatory/autoimmune disorders. We recently described the direct pathological role of extracellular Hsp70 in epidermolysis bullosa acquisita (EBA), an anti-type VII collagen autoantibody-mediated autoimmune blistering skin disease. Here, we determined the role of anti-Hsp70 autoantibodies in EBA. We observed that circulating anti-Hsp70 IgG autoantibodies were significantly elevated in EBA patients compared to healthy individuals and positively correlated with serum levels of pro-inflammatory interferon gamma (IFN-γ). The pathophysiological relevance of anti-Hsp70 IgG autoantibodies was demonstrated in an antibody transfer-induced EBA mouse model in which elevated serum levels of anti-Hsp70 IgG were found. In addition, anti-Hsp70 IgG-treated animals had a more intense clinical and histological disease activity, as well as upregulated nuclear factor kappa B (NF-κB) activation in skin biopsies compared to isotype-treated animals. Our results suggest that autoantibodies to Hsp70 may contribute to EBA development via enhanced neutrophil infiltration to the skin and activation of the NF-κB signaling pathway in an IFN-γ-associated manner.

Evidence for a role of extracellular heat shock protein 70 in epidermolysis bullosa acquisita.

Heat shock protein 90 (Hsp90) and Hsp70 are chaperones implicated in different inflammatory disorders, given their property to impact innate and adaptive immune responses. Here, we determined the so far unknown role of extracellular Hsp70 in epidermolysis bullosa acquisita (EBA), an anti-type VII collagen autoantibody-mediated blistering dermatosis. The in vivo pathophysiological relevance of extracellular Hsp70 was demonstrated in an anti-type VII collagen antibody transfer-induced EBA mouse model in which elevated blood levels of this chaperone were recorded. We found that Hsp70-treated mice had a more intense clinical disease severity compared to controls that were paralleled by increased levels of cutaneous matrix metalloproteinase 9 and plasma hydrogen peroxide. The latter finding was confirmed in an independent reactive oxygen species release assay using EBA-specific immune complexes combined with recombinant Hsp70. Finally, cell culture experiments using human naive peripheral blood mononuclear cells (PBMC) revealed that extracellular Hsp70 stimulated the secretion of the T cell-derived pro-inflammatory cytokines IL-6 and IL-8. This work extends knowledge about the role of Hsps in autoimmune bullous diseases, suggesting that extracellular Hsp70 represents a pathophysiological factor and potential treatment target in EBA.

The Effect of Different Agrotechnical Treatments on the Establishment of Miscanthus Hybrids in Soil Contaminated with Trace Metals.

Climate change and man-made pollution can have a negative impact on the establishment of Miscanthus plants in the field. This is particularly important because biomass can be produced on marginal land without conflicting with food crops. The establishment success depends on the hybrid chosen, the cultivation method, the climatic conditions, and the concentration of pollutants in the soil. There are several ways to increase the survival rate of the plants during the first growing season and after the first winter. One of them is the application of biochar and photodegradable plastic mulch, which can provide a solution for soils polluted with trace elements (TMEs). The aim of this study was to investigate the application of plastic mulch and biochar separately and in combination at the planting stage for two Miscanthus hybrids planted by the rhizome method (TV1) and seedling plugs (GNT43) on soils contaminated with trace metal elements (Pb, Cd, Zn). TV1 seems unsuitable for TME-contaminated field cultivation, as the survival rate was <60% in most treatments studied. The selected treatments did not increase the survival rate. Furthermore, the application of plastic mulch in combination with biochar resulted in a significant reduction of this parameter, regardless of the hybrid studied. The applied agrotechnology did not influence the TME accumulation in the aboveground plant parts in TV1, while Pb and Cd in GNT43 showed significantly higher values in all treatments. Contrary to expectations, biochar and plastic mulch applied separately and together neither increased survival nor reduced the accumulation of toxic TMEs during establishment on soil contaminated with TMEs and after the first growing season.

Circulating heat shock protein 90 (Hsp90) and autoantibodies to Hsp90 are increased in patients with atopic dermatitis.

Atopic dermatitis (AD) is one of the most common chronic inflammatory dermatoses characterized by persistent itching and recurrent eczematous lesions. While the primary events and key drivers of AD are topics of ongoing debate, cutaneous inflammation due to inappropriate IgE (auto)antibody-related immune reactions is frequently considered. Highly conserved and immunogenic heat shock protein 90 (Hsp90), a key intra- and extracellular chaperone, can activate the immune response driving the generation of circulating anti-Hsp90 autoantibodies that are found to be elevated in several autoimmune disorders. Here, for the first time, we observed that serum levels of Hsp90 and anti-Hsp90 IgE autoantibodies are significantly elevated (p < 0.0001) in AD patients (n = 29) when compared to age- and gender-matched healthy controls (n = 70). We revealed a positive correlation (0.378, p = 0.042) between serum levels of Hsp90 and the severity of AD assessed by Scoring Atopic Dermatitis (SCORAD). In addition, seropositivity for anti-Hsp90 IgE has been found in 48.27% of AD patients and in 2.85% of healthy controls. Although further studies on a larger group of patients are needed to confirm presented data, our results suggest that extracellular Hsp90 and autoantibodies to Hsp90 deserve attention in the study of the mechanisms that promote the development and/or maintenance of atopic dermatitis.

The Different Faces of Arabidopsis arenosa-A Plant Species for a Special Purpose.

The following review article collects information on the plant species Arabidopsis arenosa. Thus far, A. arenosa has been known as a model species for autotetraploidy studies because, apart from diploid individuals, there are also tetraploid populations, which is a unique feature of this Arabidopsis species. In addition, A arenosa has often been reported in heavy metal-contaminated sites, where it occurs together with a closely related species A. halleri, a model plant hyperaccumulator of Cd and Zn. Recent studies have shown that several populations of A. arenosa also exhibit Cd and Zn hyperaccumulation. However, it is assumed that the mechanism of hyperaccumulation differs between these two Arabidopsis species. Nevertheless, this phenomenon is still not fully understood, and thorough research is needed. In this paper, we summarize the current state of knowledge regarding research on A. arenosa.

Is the Age of Novel Ecosystem the Factor Driving Arbuscular Mycorrhizal Colonization in Poa compressa and Calamagrostis epigejos?

Some sites transformed or created by humans (novel ecosystem) are different both in vegetation and ecosystems establishment and development. The unknown habitat conditions and new species composition is resulting in new abiotic and biotic systems. To improve the understanding of the process governing the relationships between the environmental factors, plant species assemblages and their arbuscular mycorrhizal fungi (AMF) inoculation were studied in chronosequence on post-coal mine heaps. We hypothesized that AMF root colonization will be dependent on the age of heap and not on the dominant plant species (vegetation type). The high frequency of mycorrhizal colonization of roots (F%) of Poa compressa- and Calamagrostis epigejos-dominated vegetation type was stated. All mycorrhizal parameters were lower in C. epigejos roots when compared to P. compressa (ranging from 60% to 90%). The highest relative mycorrhizal intensity, M%, and mean abundance of arbuscula, A%, in the roots of both examined plants were recorded in vegetation patches dominated by Daucus carota. Positive and statistically significant correlations were found between F%, M%, and A%, and lack of correlation between the heaps' age and mycorrhizal parameters, and statistically significant correlations between A% and potassium and magnesium content were revealed. The interspecific relations in the novel ecosystems become more complex along with the increase of diversity.

Autoantibodies to heat shock protein 60, 70, and 90 are not altered in the anti-SARS-CoV-2 IgG-seropositive humans without or with mild symptoms.

Highly conserved heat shock proteins (Hsps) are localized in the cytoplasm and cellular organelles, and act as molecular chaperones or proteases. Members of Hsp families are released into the extracellular milieu under both normal and stress conditions. It is hypothesized that the severe acute respiratory syndrome corona virus 2 (SARS-CoV-2) has the potential to elicit autoimmunity due to molecular mimicry between human extracellular Hsps and immunogenic proteins of the virus. To confirm the above hypothesis, levels of circulating autoantibodies directed to the key human chaperones i.e., Hsp60, Hsp70, and Hsp90 in the anti-SARS-CoV-2 IgG-seropositive participants have been evaluated. Twenty-six healthy volunteers who got two doses of the mRNA vaccine encoding the viral spike protein, anti-SARS-CoV-2 IgG-positive participants (n = 15), and healthy naïve (anti-SARS-CoV-2 IgG-negative) volunteers (n = 51) have been included in this study. We found that the serum levels of anti-Hsp60, anti-Hsp70, and anti-Hsp90 autoantibodies of the IgG, IgM, or IgA isotype remained unchanged in either the anti-COVID-19-immunized humans or the anti-SARS-CoV-2 IgG-positive participants when compared to healthy naïve volunteers, as measured by enzyme-linked immunosorbent assay. Our results showing that the humoral immune response to SARS-CoV-2 did not include the production of anti-SARS-CoV-2 antibodies that also recognized extracellular heat shock protein 60, 70, and 90 represent a partial evaluation of the autoimmunity hypothesis stated above. Further testing for cell-based immunity will be necessary to fully evaluate this hypothesis.

Ecophysiology of Arabidopsis arenosa, a new hyperaccumulator of Cd and Zn.

Arabidopsis arenosa is a pseudo-metallophyte, closely related to the model hyperaccumulator of Cd and Zn Arabidopsis halleri. A. arenosa occurs naturally in both diploid (2C) and tetraploid (4C) form, in contrast to A. halleri in which only diploid forms were found. Moreover, A. arenosa similarly to A. halleri often occupies heavy metal (HM) contaminated sites. Nevertheless, knowledge about the ecophysiology of this species is very limited. Therefore, we examined fourteen populations of A. arenosa of different ploidy from Central Europe in situ, focusing on photosynthetic efficiency, pigment content and ability to accumulate selected elements. The presented results indicate that several tetraploid populations exhibit the features of Cd and Zn hyperaccumulation. On the one hand, we noted differences in physiological parameters between the studied populations, on the other, harshness of the environment caused similar physiological response such as high HM pollution. All these features suggest that A. arenosa, especially as a new hyperaccumulator of Cd and Zn and autopolyploidyzation model, may be considered a very interesting research object, particularly when investigating the mechanisms of HMs accumulation and tolerance in plants.

Effect of Drought and Heavy Metal Contamination on Growth and Photosynthesis of Silver Birch Trees Growing on Post-Industrial Heaps.

Silver birch trees (Betula pendula Roth) are a pioneering species in post-industrial habitats, and have been associated with an expansive breeding strategy and low habitat requirements. We conducted ecophysiological and dendroclimatological studies to check whether there are any features of which the modification enables birch trees to colonise extreme habitats successfully. We characterised the efficiency of the photosynthetic apparatus, the gas exchange, the content of pigments in leaves, and the growth (leaf thickness and tree-ring width) of birch trees on a post-coal mine heap, a post-smelter heap, and a reference site. Birch growth was limited mainly by temperature and water availability during summer, and the leaves of the birch growing on post-industrial heaps were significantly thicker than the reference leaves. Moreover, birch trees growing on heaps were characterised by a significantly higher content of flavonols and anthocyanins in leaves and higher non-photochemical quenching. In addition, birches growing on the post-coal mine heap accumulated a concentration of Mn in their leaves, which is highly toxic for most plant species. Increasing the thickness of leaves, and the content of flavonols and anthocyanins, as well as efficient non-photochemical quenching seem to be important features that improve the colonization of extreme habitats by birches.

Cuticular waxes-A shield of barley mutant in CBP20 (Cap-Binding Protein 20) gene when struggling with drought stress.

CBP20 (Cap-Binding Protein 20) encodes a small subunit of nuclear Cap-Binding Complex (nCBC) that together with CBP80 binds mRNA cap. We previously described barley hvcbp20.ab mutant that demonstrated higher leaf water content and faster stomatal closure than the WT after drought stress. Hence, we presumed that the better water-saving mechanism in hvcbp20.ab may result from the lower permeability of epidermis that together with stomata action limit the water evaporation under drought stress. We asked whether hvcbp20.ab exhibited any differences in wax load on the leaf surface when subjected to drought in comparison to WT cv. 'Sebastian'. To address this question, we investigated epicuticular wax structure and chemical composition under drought stress in hvcbp20.ab mutant and its WT. We showed that hvcbp20.ab mutant exhibited the increased deposition of cuticular wax. Moreover, our gene expression results suggested a role of HvCBP20 as a negative regulator of both, the biosynthesis of waxes at the level of alkane-forming, and waxes transportation. Interestingly, we also observed increased wax deposition in Arabidopsis cbp20 mutant exposed to drought, which allowed us to describe the CBP20-regulated epicuticular wax accumulation under drought stress in a wider evolutionarily context.

Different strategies of Cd tolerance and accumulation in Arabidopsis halleri and Arabidopsis arenosa.

Pseudometallophytes are commonly used to study the evolution of metal tolerance and accumulation traits in plants. Within the Arabidopsis genus, the adaptation of Arabidopsis halleri to metalliferous soils has been widely studied, which is not the case for the closely related species Arabidopsis arenosa. We performed an in-depth physiological comparison between the A. halleri and A. arenosa populations from the same polluted site, together with the geographically close non-metallicolous (NM) populations of both species. The ionomes, growth, photosynthetic parameters and pigment content were characterized in the plants that were growing on their native site and in a hydroponic culture under Cd treatments. In situ, the metallicolous (M) populations of both species hyperaccumulated Cd and Zn. The NM population of A. halleri hyperaccumulated Cd and Zn while the NM A. arenosa did not. In the hydroponic experiments, the NM populations of both species accumulated more Cd in their shoots than the M populations. Our research suggests that the two Arabidopsis species evolved different strategies of adaptation to extreme metallic environments that involve fine regulation of metal homeostasis, adjustment of the photosynthetic apparatus and accumulation of flavonols and anthocyanins.