Physiological response of adult Salix aurita in wetland vegetation affected by flooding with As-rich fine pyrite particles.

An uncontrolled, natural episode of flooding with waters contaminated with As-rich pyrite (FeAsS) particles caused serious ecological damage leading to necrosis of plants growing in a fresh wet meadow located in an area characterized by unique geological structures rich in arsenopyrites. One of the few plant species capable of surviving this event was Salix aurita L., which grew in numbers in the analyzed area, but individual plants were affected differently by toxic flooding. No significant phenotypic changes (Group I), through partial leaf and/or stem necrosis (Group II) up to necrosis of the whole parental plant and root suckers (Group III), were observed for various willow clumps. These varied phenotypic responses of S. aurita to As-rich sediments were compared with the biochemical status of the foliage of willow trees, and with their rhizosphere physiological parameters. Our in situ study revealed that the biochemical status of leaves reflects the phenotypic damage incurred by adult willows growing in their natural environment and affected by the flooding. In leaves of willows with increasingly negative phenotypic changes (Groups I → II → III) as well as increasing levels of reactive oxygen species, malondialdehyde and decreased levels of glutathione and thiol groups were detected. Phytochelatins, commonly considered major As chelators, were not detected in S. aurita leaves. Despite a decrease in the size of leaves with the intensity of tree damage, all leaves expressed a normal level of leaf pigments. Phenotypic changes observed for particular willow clumps were only partly related to soil As levels. Moreover, As and S (but not Fe) foliar levels were related but did not correspond strictly with foliar biochemical features, or with soil As levels, soil pH or soil microbial activity, with the latter two drastically decreased in the rhizospheres of willows from Groups II and III.

Arsenic uptake, speciation and physiological response of tree species (Acer pseudoplatanus, Betula pendula and Quercus robur) treated with dimethylarsinic acid.

The aim of the study was to evaluate the effect of dimethylarsinic acid (DMA) on growth parameters and levels of stress-related metabolites in Acer pseudoplatanus, Betula pendula and Quercus robur. The increase of DMA concentration in the solution led to a notable growth retardation of trees. An intense As accumulation (mainly As(III) and As(V)) expressed as BCF and TF > 1 was recorded only for Q. robur. Generally a decrease in contents of cellulose, hemicellulose and holocellulose with a simultaneous increase in lignin content were recorded. Phenolic composition of leaf extracts was modified by DMA, while root and rhizosphere extracts were poor in phenolics. Toxicity of DMA leads to a significant drop in salicylic acid content in leaves observed at lower doses. Higher DMA levels caused a second, probably ROS-derived depletion of the metabolite accompanied with a severe growth retardation, most pronounced in the case of B. pendula. DMA caused the inhibition of LMWOA biosynthesis in roots of A. pseudoplatanus, B. pendula and their exudation into the rhizosphere, while in Q. robur roots and leaves a stimulation of their accumulation was observed. Disturbances in the activity of enzymatic antioxidants were observed for all the species following the increasing level of DMA.

Combined use of companion planting and PGPR for the assisted phytoextraction of trace metals (Zn, Pb, Cd).

Biomass production and metal accumulation in plant tissue (bioconcentration) are two critical factors limiting the phytoextraction rate. Metal translocation to aboveground organs should be accounted for as the third most important factor, as harvesting of the plant roots is usually economically disadvantageous. These three parameters could be potentially increased with the use of companion planting, a well-known agricultural technique, and inoculation with plant growth-promoting bacteria (PGPB). The aim of the study was to determine whether intercropping and inoculation with endophytic PGPB (Burkholderia phytofirmans PsJNT) can increase the efficiency of phytoextraction of Zn, Pb, and Cd. The study was conducted on Brassica juncea (L.) Czern. "Malopolska" grown in a monoculture or co-planted with Zea mays L. "Codimon" and Medicago sativa L. "Sanditi." Results show that companion planting and inoculation with rhizobacteria can increase the efficiency of metal phytoextraction, mainly by increasing the yield of dry biomass and the survival rate of plants grown on contaminated soil. We have shown that the simultaneous planting of B. juncea with M. sativa and inoculation with PGPB were the most efficient variants of assisted phytoextraction reaching a recovery of 95% Zn, 90% Cd, and on average about 160% Pb compared with control B. juncea plants grown in monoculture.

Differences of Acer platanoides L. and Tilia cordata Mill. Response patterns/survival strategies during cultivation in extremely polluted mining sludge - A pot trial.

The study aimed to evaluate the physiological mechanisms underlying differences in metals and metalloid uptake and tolerance of two tree species cultivated in mining waste material. Two-year old Acer platanoides L. and Tilia cordata Mill. were cultivated in mining sludge characterized by high pH, salinity and an extremely high concentration of As. Both species were able to develop leaves from leafless seedlings, however, their total biomass was greatly reduced in comparison to control plants, following the severe disturbances in chlorophyll content. Phytoextraction abilities were observed for T. cordata for Ba, Nb, Rb and Se, and phytostabilisation was stated for Pd, Ru, Sc and Sm for both species, Ba and Nd for A. platonoides and Be for T. cordata only. Metal exclusion was observed for the majority of detected elements indicating an intense limitation of metal transport to photosynthetic tissue. A diversified uptake of elements was accompanied by a species-specific pattern of physiological reaction during the cultivation in sludge. Organic ligands (glutatnione and low-molecular-weight organic acids) were suppressed in A. platanoides, and enhanced biosynthesis of phenolic compounds was observed for both species, being more pronounced in T. cordata. Despite its higher accumulation of key metabolites for plant reaction to oxidative stress, such as phenolic acids, flavonoids and organic ligands, T. cordata exhibited relatively lower tolerance to sludge, probably due to the increased uptake and translocation rate of toxic metal/loids to aerial organs and/or restricted accumulation of salicylic acid which is known to play a decisive role in mechanisms of plant tolerance.

Effects of binary metal combinations on zinc, copper, cadmium and lead uptake and distribution in Brassica juncea.

The interaction between lead, copper, cadmium and zinc in their binary combinations was investigated in Indian mustard seedlings (Brassica juncea L. var. Malopolska). Fourteen-days-old seedlings were treated with Pb(NO3)2, CuSO4, CdCl2, ZnSO4 at 50µmol of metal ion concentration and at 25µmol of each metal ion in combinations. Metal combinations were generally more inhibiting in terms of biomass production. This inhibiting effect followed an order: Cu+Cd>Cu+Zn, Cd+Pb>Cu+Pb>Zn+Pb, Cu>Cd>Zn>Zn+Cd>Pb. We observed synergistic and antagonistic effects of metal uptake in binary metal treatments, suggesting metal crosstalk at the plant uptake site. Metal content in plant tissues varied among different combinations. The metal concentrations followed an order of Pb>Cu>Zn>Cd in roots, Zn>Cu>Pb>Cd in the stem and Zn>Cu>Cd>Pb in leaves. Presence of metals altered the distribution of micronutrients (Cu, Zn) in plants: Cu concentration was lowered in roots and leaves and increased in stems; Zn content was increased in plants, with stems having up to 4 or 5 times more Zn than in control plants.

Rhizoremediation of diesel-contaminated soil with two rapeseed varieties and petroleum degraders reveals different responses of the plant defense mechanisms.

Plant-assisted bioremediation (rhizoremediation) stands out as a potential tool to inactivate or completely remove xenobiotics from the polluted environment. Therefore, it is of key importance to find an adequate combination of plant species and microorganisms that together enhance the clean-up process. To understand the response of plants upon bioaugmentation, the antioxidative and detoxification system was analyzed in high and low erucic acid rapeseed varieties (HEAR and LEAR, respectively), after 8 weeks of their treatment with petroleum degraders and 6000 mg diesel oil/kg dry soil. The oxidative stress was enhanced in LEAR being exposed to sole diesel oil, in comparison with HEAR. However, when LEAR plants were additionally inoculated with bacteria, suppression of total catalase (CAT) and ascorbate peroxidase (APX) activity were observed. Interestingly, glutathione transferase (GST) activity was found in these plants at a much higher level than in HEAR, which correlated with a more efficient diesel removal performed by LEAR in the polluted soil and upon bioaugmentation. A distinct profile of polycyclic aromatic hydrocarbons (PAH) was detected in leaves of these plants. Neither LEAR nor HEAR experienced any changes in the photosynthetic capacity upon diesel pollution and presence of petroleum degraders, which supports the usefulness of rhizoremediation with rapeseed.

Response of the pea roots defense systems to the two-element combinations of metals (Cu, Zn, Cd, Pb).

The presence of the single metals (Cd, Pb, Cu, Zn) induces ROS (reactive oxygen species) production and causes oxidative stress in plants. While applied in two-element combinations, trace metals impact organisms in a more complex way. To assess the resultant effect we treated the pea grown hydroponically with the trace metals in variants: CuPb, CuCd, CuZn, PbCd, ZnPb, ZnCd in concentrations of 25 µM for each metal ion. Abiotic stress inhibited root elongation growth, decreased biomass production, induced changes in root colour and morphology. It changed rate of ROS production, malondialdehyde content, increased activity and altered gene expression of defence enzymes (superoxide dysmutase, catalase, ascorbate peroxidase, glutathione reductase, γ-glutamylcysteine synthetase).

Arabidopsis protein phosphatase 2C ABI1 interacts with type I ACC synthases and is involved in the regulation of ozone-induced ethylene biosynthesis.

Ethylene plays a crucial role in various biological processes and therefore its biosynthesis is strictly regulated by multiple mechanisms. Posttranslational regulation, which is pivotal in controlling ethylene biosynthesis, impacts 1-aminocyclopropane 1-carboxylate synthase (ACS) protein stability via the complex interplay of specific factors. Here, we show that the Arabidopsis thaliana protein phosphatase type 2C, ABI1, a negative regulator of abscisic acid signaling, is involved in the regulation of ethylene biosynthesis under oxidative stress conditions. We found that ABI1 interacts with ACS6 and dephosphorylates its C-terminal fragment, a target of the stress-responsive mitogen-activated protein kinase, MPK6. In addition, ABI1 controls MPK6 activity directly and by this means also affects the ACS6 phosphorylation level. Consistently with this, ozone-induced ethylene production was significantly higher in an ABI1 knockout strain (abi1td) than in wild-type plants. Importantly, an increase in stress-induced ethylene production in the abi1td mutant was compensated by a higher ascorbate redox state and elevated antioxidant activities. Overall, the results of this study provide evidence that ABI1 restricts ethylene synthesis by affecting the activity of ACS6. The ABI1 contribution to stress phenotype underpins its role in the interplay between the abscisic acid (ABA) and ethylene signaling pathways.

Determination of cadmium and lead species and phytochelatins in pea (Pisum sativum) by HPLC-ICP-MS and HPLC-ESI-MSn.

An analytical approach based on hyphenated techniques was used for studying the speciation of cadmium and lead in Pisum sativum. Proper preservation conditions were employed to avoid the oxidation of -SH groups and corresponding decomposition of metal-binding complexes. SEC column was washed with 5 mM beta-mercaptoethanol and then samples were analysed using ICP-MS as a detector. Results showed that cadmium is the inhibitor of lead uptake. HPLC-ESI-MS(n) assays revealed fragmentation pathways of phytochelatins.

Enhancing phytoremediative ability of Pisum sativum by EDTA application.

The aim of our research was to demonstrate how the presence of EDTA affects resistance of pea plants to Pb and Pb-EDTA presence, and to show the effectivity of lead ions accumulation and translocation. It was determined that EDTA not only increased the amount of Pb taken up by plants but also Pb ion transport through the xylem and metal translocation from roots to stems and leaves. It can be seen in the presented research results that addition of the chelator with Pb limited metal phytotoxicity. We also demonstrated a significant effect of EDTA not only on Pb accumulation and metal transport to the aboveground parts but also on the profile and amount of thiol compounds: glutathione (GSH), homoglutathione (hGSH) or phytochelatins (PCs), synthesized by the plants. We observed a significant effect of the synthetic chelator on increasing the level of Pb accumulation in roots of plants treated with Pb including EDTA (0.5 and 1 mM). Pisum sativum plants treated only with 1 mM Pb(NO3)2 accumulated over 50 mg Pb x g(-1) dry wt during 4 days of cultivation. Whereas in roots of pea plants exposed to Pb+0.5 mM EDTA 35% more Pb was observed. When 1 mM EDTA was applied roots of pea accumulated over 67% more metal. The presence of EDTA also increased metal uptake and transport to the aboveground parts. In pea plants treated only with 1 mM lead nitrate less than 3 mg Pb x g(-1) dry wt was transported, whereas in P. sativum treated with Pb-EDTA doubled amount of Pb was observed in stems and leaves.

Accumulation and detoxification of lead ions in legumes.

This study focuses on lead accumulation in roots, stems and leaves of three plant species of the Fabacea family: Vicia faba, Pisum sativum and Phaseolus vulgaris grown hydroponically in a medium supplemented with 1 mM concentration of lead. The largest amount of lead, up to 75 mg Pb/g dry weight, was accumulated in roots of P. vulgaris. The highest rate of Pb ions uptake from the medium took place during the first 10 h of incubation with lead and after 96 h of incubation lead content in the medium decreased by half. Thus, it was suggested that P. vulgaris could be used in rhizofiltration--the use of plant roots to absorb pollutants from water contaminated with lead. At the same time we studied the influence of lead on acid soluble thiol, glutathione, homoglutathione contents and the synthesis of phyto- and homophytochelatins in roots of V. faba, P. sativum and P. vulgaris grown hydroponically. Activation of the detoxicative-phytochelatin system was observed in the cytosol of root cells of the tested plants. This system was composed of phytochelatins (PCs) in roots of V. faba, homophytochelatins (hPCs) in P. vulgaris roots and both PCs and hPCs in P. sativum roots. The total content of PCs and hPCs in roots of P. sativum was very high and reached around 4800 (expressed in nmol SH x g(-1)FW) and induction of their synthesis occurred after only 2 h of treatment with 1 mM Pb.