Arsenic accumulation and speciation in two cultivars of Pteris cretica L. and characterization of arsenate reductase PcACR2 and arsenite transporter PcACR3 genes in the hyperaccumulating cv. Albo-lineata.

Pollution and poisoning with carcinogenic arsenic (As) is of major concern globally. Interestingly, there are ferns that can naturally tolerate remarkably high As concentrations in soils while hyperaccumulating this metalloid in their fronds. Besides Pteris vittata in which As-related traits and molecular determinants have been studied in detail, the As hyperaccumulation status has been attributed also to Pteris cretica. We thus inspected two P. cretica cultivars, Parkerii and Albo-lineata, for As hyperaccumulation traits. The cultivars were grown in soils supplemented with 20, 100, and 250 mg kg-1 of inorganic arsenate (iAsV). Unlike Parkerii, Albo-lineata was confirmed to be As tolerant and hyperaccumulating, with up to 1.3 and 6.4 g As kg-1 dry weight in roots and fronds, respectively, from soils amended with 250 mg iAsV kg-1. As speciation analyses rejected that organoarsenical species and binding with phytochelatins and other proteinaceous ligands would play any significant role in the biology of As in either cultivar. While in Parkerii, the dominating As species, particularly in roots, occurred as iAsV, in Albo-lineata the majority of the root and frond As was apparently converted to iAsIII. Parkerii markedly accumulated iAsIII in its fronds when grown on As spiked soils. Considering the roles iAsV reductase ACR2 and iAsIII transporter ACR3 may have in the handling of iAs, we isolated Albo-lineata PcACR2 and PcACR3 genes closely related to P. vittata PvACR2 and PvACR3. The gene expression analysis in Albo-lineata fronds revealed that the transcription of PcACR2 and PcACR3 was clearly As responsive (up to 6.5- and 45-times increase in transcript levels compared to control soil conditions, respectively). The tolerance and uptake assays in yeasts showed that PcACRs can complement corresponding As-sensitive mutations, indicating that PcACR2 and PcACR3 encode functional proteins that can perform, respectively, iAsV reduction and membrane iAsIII transport tasks in As-hyperaccumulating Albo-lineata.

Effect of arsenic stress on 5-methylcytosine, photosynthetic parameters and nutrient content in arsenic hyperaccumulator Pteris cretica (L.) var. Albo-lineata.

BACKGROUND: Arsenic toxicity induces a range of metabolic responses in plants, including DNA methylation. The focus of this paper was on the relationship between As-induced stress and plant senescence in the hyperaccumulator Pteris cretica var. Albo-lineata (Pc-Al). We assume difference in physiological parameters and level of DNA methylation in young and old fronds as symptoms of As toxicity. RESULTS: The As accumulation of Pc-Al fronds, grown in pots of haplic chernozem contaminated with 100 mg As kg- 1 for 122 days, decreased with age. Content of As was higher in young than old fronds for variants with 100 mg As kg- 1 (2800 and 2000 mg As kg- 1 dry matter, respectively). The highest As content was determined in old fronds of Pc-Al grown in pots with 250 mg As kg- 1. The increase with age was confirmed for determined nutrients - Cu, Mg, Mn, S and Zn. A significant elevation of all analysed nutrients was showed in old fronds. Arsenic accumulation affected DNA methylation status in fronds, but content of 5-methylcytosine (5mC) decreased only in old fronds of Pc-Al (from 25 to 12%). Determined photosynthetic processes showed a decrease of fluorescence, photosynthetic rate and chlorophylls of As treatments in young and old fronds. Water potential was decreased by As in both fronds. Thinning of the sclerenchymatous inner cortex and a reduction in average tracheid metaxylem in the vascular cylinder was showed in roots of As treatment. Irrespective to fronds age, physiological parameters positively correlated with a 5mC while negatively with direct As toxicity. Opposite results were found for contents of Cu, Mg, Mn, S and Zn. CONCLUSIONS: The results of this paper point to changes in the metabolism of the hyperaccumulator plant Pc-Al, upon low and high exposure to As contamination. The significant impact of As on DNA methylation was found in old fronds. Irrespective to fronds age, significant correlations were confirmed for 5mC and As toxicity. Our analysis of the very low water potential values and lignification of cell walls in roots showed that transports of assimilated metabolites and water between roots and fronds were reduced. As was showed by our results, epigenetic changes could affect studied parameters of the As hyperaccumulator plant Pc-Al, especially in old fronds.

Multicontamination Toxicity Evaluation in the Model Plant Lactuca sativa L.

Many contaminated soils contain several toxic elements (TEs) in elevated contents, and plant-TE interactions can differ from single TE contamination. Therefore, this study investigated the impact of combined contamination (As, Cd, Pb, Zn) on the physiological and metabolic processes of lettuce. After 45 days of exposure, TE excess in soil resulted in the inhibition of root and leaf biomass by 40 and 48%, respectively. Oxidative stress by TE accumulation was indicated by markers-malondialdehyde and 5-methylcytosine-and visible symptoms of toxicity (leaf chlorosis, root browning) and morpho-anatomical changes, which were related to the change in water regime (water potential decrease). An analysis of free amino acids (AAs) indicated that TEs disturbed N and C metabolism, especially in leaves, increasing the total content of free AAs and their families. Stress-induced senescence by TEs suggested changes in gas exchange parameters (increase in transpiration rate, stomatal conductance, and intercellular CO2 concentration), photosynthetic pigments (decrease in chlorophylls and carotenoids), a decrease in water use efficiency, and the maximum quantum yield of photosystem II. These results confirmed that the toxicity of combined contamination significantly affected the processes of lettuce by damaging the antioxidant system and expressing higher leaf sensitivity to TE multicontamination.

Arsenic Soil Contamination and Its Effects on 5-Methylcytosine Levels in Onions and Arsenic Distribution and Speciation.

Arsenic represents a serious health threat in localities with a high arsenic-polluted environment and can easily get into the human food chain through agronomy production in areas affected by arsenic contamination. Onion plants that were grown in controlled conditions in arsenic-contaminated soil (5, 10, and 20 ppm) were harvested 21 days after contamination. Arsenic levels (from 0.43 ± 0.03 µg g-1 to 1761.11 ± 101.84 µg g-1) in the onion samples were high in the roots and low in the bulbs and leaves, which is probably caused by a reduced ability of the onions to transport arsenic from roots to bulbs and leaves. Arsenic species As(V) and As(III) in As(V)-contaminated soil samples were represented strongly in favor of the As(III) species. This indicates the presence of arsenate reductase. Levels of 5-methylcytosine (5-mC) (from 5.41 ± 0.28% to 21.17 ± 1.33%) in the onion samples were also higher in the roots than in the bulbs and leaves. Microscopic sections of the roots were examined, and the most damage was found in the 10 ppm As variant. Photosynthetic parameters pointed to a significant decrease in photosynthetic apparatus activity and the deterioration of the physiological state of plants as arsenic content increased in the soil.

Effect of Arsenic Soil Contamination on Stress Response Metabolites, 5-Methylcytosine Level and CDC25 Expression in Spinach.

Experimental spinach plants grown in soil with (5, 10 and 20 ppm) arsenic (As) contamination were sampled in 21 days after As(V) contamination. Levels of As in spinach samples (from 0.31 ± 0.06 µg g-1 to 302.69 ± 11.83 µg g-1) were higher in roots and lower in leaves, which indicates a low ability of spinach to translocate As into leaves. Species of arsenic, As(III) and As(V), were represented in favor of the As (III) specie in contaminated variants, suggesting enzymatic arsenate reduction. In relation to predominant As accumulation in roots, changes in malondialdehyde levels were observed mainly in roots, where they decreased significantly with growing As contamination (from 11.97 ± 0.54 µg g-1 in control to 2.35 ± 0.43 µg g-1 in 20 ppm As). Higher values in roots than in leaves were observed in the case of 5-methylcytosine (5-mC). Despite that, a change in 5-mC by As contamination was further deepened in leaves (from 0.20 to 14.10%). In roots of spinach, expression of the CDC25 gene increased by the highest As contamination compared to the control. In the case of total phenolic content, total flavonoid content, total phenolic acids content and total antioxidant capacity were higher levels in leaves in all values, unlike the roots.

Leaf fitness and stress response after the application of contaminated soil dust particulate matter.

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.

Tocotrienols and tocopherols in colored-grain wheat, tritordeum and barley.

Colored-grain spring and winter wheat, spring tritordeum and barley (blue aleurone, purple pericarp, and yellow endosperm) from the harvests 2014 and 2015 were evaluated for tocol contents by HPLC-FD. Higher content of total tocols was found in spring wheat varieties compared with winter varieties. Four tocols (ß-tocotrienol, α-tocotrienol, ß-tocopherol, and α-tocopherol) were identified in wheat and tritordeum varieties. Dominant tocols in purple- and blue-grained wheat and yellow-grained tritordeum were α-tocopherol and ß-tocotrienol, whereas spring barley varieties differed from wheat and tritordeum by high α-tocotrienol content. Tocol content was significantly affected by genotype and in a lesser extent in some varieties and lines also by rainfall and temperatures during crop year. Higher rainfall and lower temperatures caused in most varieties higher tocol contents. Purple- and blue-grained wheat lines with higher tocol, anthocyanin and phenolic acids with health benefits may be useful for breeding new varieties.