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.

Response of Carrot (Daucus carota L.) to Multi-Contaminated Soil from Historic Mining and Smelting Activities.

A pot experiment was undertaken to investigate the effect of Cd, Pb and Zn multi-contamination on the physiological and metabolic response of carrot (Daucus carota L.) after 98 days of growth under greenhouse conditions. Multi-contamination had a higher negative influence on leaves (the highest Cd and Zn accumulation) compared to the roots, which showed no visible change in terms of anatomy and morphology. The results showed the following: (i) significantly higher accumulation of Cd, Zn, and Pb in the multi-contaminated variant (Multi) compared to the control; (ii) significant metabolic responses-an increase in the malondialdehyde content of the Multi variant compared to the control in the roots (by 20%), as well as in the leaves (by 53%); carotenoid content in roots decreased by 31% in the Multi variant compared with the control; and changes in free amino acids, especially those related to plant stress responses. The determination of hydroxyproline and sarcosine may reflect the higher sensitivity of carrot leaves to multi-contamination in comparison to roots. A similar trend was observed for the content of free methionine (significant increase of 31% only in leaves); (iii) physiological responses (significant decreases in biomass, changes in gas-exchange parameters and chlorophyll a); and (iv) significant changes in enzymatic activities (chitinase, alanine aminopeptidase, acid phosphatase) in the root zone.

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.

Health Risk and Quality Assessment of Vegetables Cultivated on Soils from a Heavily Polluted Old Mining Area.

Three garden vegetables-radish, carrot and lettuce-were cultivated in a pot experiment using two soils from the Príbram area polluted mainly by cadmium (Cd), zinc (Zn), lead (Pb) and chromium (Cr). The soils of the Príbram district, Czech Republic, are heavily polluted as a result of the atmospheric deposition of toxic elements originating from historic lead-silver mining and smelting activities. The results showed that lettuce absorbed the highest amounts of toxic elements (Cd 28 and 30, Cr 12 and 13, Zn 92 and 205 mg·kg-1 DW), except Pb, which was higher in radish (30 and 49 mg·kg-1 DW). Changes in macronutrient contents in edible parts were not found, except for sulfur. A higher total free amino acids (fAAs) accumulation was shown in all vegetables in more contaminated soil, with the highest fAA content being in radish. A group of essential fAAs reached 7-24% of total fAAs in vegetables. The risk to human health was characterized using the target hazard quotient and total hazard index (HI). The cumulative effect of the consumption of vegetables with HI > 1 showed possible non-carcinogenic health effects for lettuce and carrot. HI decreased in the order Cd > Pb > Cr > Zn. The carcinogenic risk of toxic elements decreased in the order Cd > Cr > Pb (0.00054, 0.00026, 0.00003). These values showed a carcinogenic risk from the consumption of lettuce and carrot and confirmed that the adult population of the studied area is at high risk if lettuce and carrot cultivated in this area are consumed daily.

Biochemical and physiological changes in Zea mays L. after exposure to the environmental pharmaceutical pollutant carbamazepine.

The presence of pharmaceuticals in the environment is a matter of great concern. They are consistently found in the environment, raising concerns regarding human exposure through dietary intake. In this study, we observed the effect of the application of carbamazepine at 0.1, 1, 10, and 1000 µg per kg of soil contamination levels to assess stress metabolism in Zea mays L. cv. Ronaldinio at the 4th leaf, tasselling, and dent phenological stages. The transfer of carbamazepine to the aboveground and root biomass was assessed, and uptake increased dose-dependently. No direct effect on biomass production was observed, but multiple physiological and chemical changes were observed. Major effects were consistently observed at the 4th leaf phenological stage for all contamination levels, including reduced photosynthetic rate, reduced maximal and potential activity of photosystem II, decreased water potential, decreased carbohydrates (glucose and fructose) and γ-aminobutyric acid in roots, and increased maleic acid and phenylpropanoids (chlorogenic acid and its isomer, 5-O-caffeoylquinic acid) in aboveground biomass. A reduction in net photosynthesis was observed for the older phenological stages, whereas no other relevant and consistent physiological and metabolic changes related to contamination exposure were detected. Our results indicate that Z. mays can overcome the environmental stress caused by the accumulation of carbamazepine with notable metabolic changes at the early phenological stage; however, older plants adapted and only exhibited minor effects in the presence of the contaminant. The potential implications for agricultural practice could be associated with the plant's response to simultaneous stresses due to metabolite changes associated with oxidative stress.

Accumulation of Toxic Arsenic by Cherry Radish Tuber (Raphanus sativus var. sativus Pers.) and Its Physiological, Metabolic and Anatomical Stress Responses.

In a pot experiment, cherry radish (Raphanus sativus var. sativus Pers. 'Viola') was cultivated under two levels of As soil contamination-20 and 100 mg/kg. The increasing As content in tubers with increasing soil contamination led to changes in free amino acids (AAs) and phytohormone metabolism and antioxidative metabolites. Changes were mainly observed under conditions of high As contamination (As100). The content of indole-3-acetic acid in tubers varied under different levels of As stress, but As100 contamination led to an increase in its bacterial precursor indole-3-acetamide. A decrease in cis-zeatin-9-riboside-5'-monophosphate content and an increase in jasmonic acid content were found in this treatment. The free AA content in tubers was also reduced. The main free AAs were determined to be transport AAs (glutamate-Glu, aspartate, glutamine-Gln, asparagine) with the main portion being Gln. The Glu/Gln ratio-a significant indicator of primary N assimilation in plants-decreased under the As100 treatment condition. A decrease in antioxidative metabolite content-namely that of ascorbic acid and anthocyanins-was observed in this experiment. A decline in anthocyanin content is related to a decrease in aromatic AA content which is crucial for secondary metabolite production. The changes in tubers caused by As contamination were reflected in anatomical changes in the radish tubers and roots.

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.

Is the harvest of Salix and Populus clones in the growing season truly advantageous for the phytoextraction of metals from a long-term perspective?

The phytoextraction potential of short-rotation field coppice plantations was investigated on soil historically contaminated with risk elements (REs), especially Cd, Pb and Zn. The main objective of the study was to assess the effect of biomass harvest time (summer harvest versus traditional winter one) on biomass yield, REs accumulation and removal in the long-term study. The precise field experiment with two Salix clones and two Populus clones was established in April 2009. Shoots of all clones were harvested in February 2012 for the first time, and then identical branches and leaves were harvested every two years in September (summer harvest = SH) and the branches every two years in February (winter harvest = WH). The first summer harvest seemed to be more promising compared to the winter one, but the yields in the second and third harvests were inconsistent. The total phytoextraction efficiency of the SH declined in second and third harvests due to a decrease of leaves/aboveground biomass ratio, and the RE concentrations in leaves. Clonal Salix smithiana was the most productive one in the SH, with a dry matter yield of 15.1 t ha-1 year-1 and showed promising extraction ability for Cd (11.65 %) and Zn (4.59 %) over a 6-year field experiment. A lower portion of Cd (6.97 %) and Zn (2.38 %) was removed by this clone in the WH (calculated from the total soil content of REs). SH was more reasonable for phytoextraction compared to WH. Higher RE concentrations were found in leaves of Salix compared to Populus. Populus accumulated the highest Pb content in the branches; unfortunately, the Pb extraction was low, due to extremely high soil Pb contamination. Locally bred willows and poplars performed substantially better than internationally recognised ones, indicating the importance of phytoremediation optimisation, including clone selection, for suitable climatic conditions.

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.

Arsenic Toxicity-Induced Physiological and Metabolic Changes in the Shoots of Pteris cretica and Spinacia oleracea.

Arsenic is a ubiquitous toxic element that can be accumulated into plant parts. The present study investigated the response of Pteris cretica and Spinacia oleracea to As treatment through the analysis of selected physiological and metabolic parameters. Plants were grown in pots in As(V) spiked soil (20 and 100 mg/kg). Plants' physiological condition was estimated through the determination of elements, gas-exchange parameters, chlorophyll fluorescence, water potential, photosynthetic pigments, and free amino acid content. The results confirmed differing As accumulation in plants, as well as in shoots and roots, which indicated that P. cretica is an As-hyperaccumulator and that S. oleracea is an As-root excluder. Variations in physiological and metabolic parameters were observed among As treatments. Overall, the results revealed a significant effect of 100 mg/kg As treatment on the analysed parameters. In both plants, this treatment affected growth, N, Mg, S, Mn, and Zn content, as well as net photosynthetic rate, chlorophyll fluorescence, and total free amino acid content. In conclusion, the results reflect the similarity between P. cretica and S. oleracea in some aspects of plants' response to As treatment, while physiological and metabolic parameter changes related to As treatments indicate the higher sensitivity of S. oleracea.

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.

Response of cytokinins and nitrogen metabolism in the fronds of Pteris sp. under arsenic stress.

Given the close relationship between cytokinins (CKs), photosynthesis and nitrogen metabolism, this study assessed the effect of arsenic (As) contamination on these metabolic components in the As-hyperaccumulators Pteris cretica L. var. Albo-lineata (Pc-A) and var. Parkerii (Pc-P) as well as the As-non-hyperaccumulator Pteris straminea Mett. ex Baker (Ps). The ferns were cultivated in a pot experiment for 23 weeks in soil spiked with As at the levels 20 and 100 mg·kg-1. For the purpose of this study, the CKs were placed into five functionally different groups according to their structure and physiological roles: bioactive forms (bCKs; CK free bases); inactive or weakly active forms (dCKs; CK N-glucosides); transport forms (tCKs; CK ribosides); storage forms (sCKs; O-glucosides); and primary products of CK biosynthesis (ppbCKs; CK nucleotides). An important finding was higher CKs total content, accumulation of sCKs and reduction of dCKs in As-hyperaccumulators in contrast to non-hyperaccumulator ferns. A significant depletion of C resources was confirmed in ferns, especially Ps, which was determined by measuring the photosynthetic rate and chlorophyll fluorescence. A fluorescence decrease signified a reduction in the C/N ratio, inducing an increase of bioactive CKs forms in Pc-P and Ps. The impact of As on N utilization was significant in As-hyperaccumulators. The glutamic acid/glutamine ratio, an indicator of primary N assimilation, diminished in all ferns with increased As level in the soil. In conclusion, the results indicate a large phenotypic diversity of Pteris species to As and suggest that the CKs composition and the glutamic acid/glutamine ratio can be used as a tool to diagnose As stress in plants.

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.

Cadmium toxicity induced contrasting patterns of concentrations of free sarcosine, specific amino acids and selected microelements in two Noccaea species.

Cadmium (Cd) toxicity affects numerous metabolic processes in plants. In the presence of Cd, plants accumulate specific amino acids which may be beneficial to developing Cd tolerance. Our study aimed to characterize the changes in the metabolism of selected free amino acids that are associated with Cd tolerance, and investigate the levels of selected microelements in order to relate these changes to the adaptation strategies of two metallophytes-Noccaea caerulescens (Redlschlag, Austria) and Noccaea praecox (Mezica, Slovenia). The plants were exposed to Cd contamination (90 mg Cd/kg soil) for 120 days in a pot experiment. Our results showed higher Cd accumulation in N. praecox compared to N. caerulescens. Cadmium contamination reduced the zinc and nickel levels in both species and a mixed effect was determined for copper and manganese content. Differences in free amino acid metabolism were observed between the two metallophytes growing under Cd-free and Cd-loaded conditions. Under Cd-free conditions, aromatic amino acids (phenylalanine, tryptophan and tyrosine) and branched-chain amino acids (leucine, isoleucine and valine) were accumulated more in the leaves of N. praecox than in N. caerulescens. Cd stress increased the content of these amino acids in both species but this increase was significant only in N. caerulescens leaves. Marked differences in the responses of the two species to Cd stress were shown for alanine, phenylalanine, threonine and sarcosine. Cadmium contamination also induced an increase of threonine as alanine and sarcosine decrease, which was larger in N. caerulescens than in N. praecox. All these factors contribute to the higher adaptation of N. praecox to Cd stress.

Regulation of odd-numbered fatty acid content plays an important part in the metabolism of the hyperaccumulator Noccaea spp. adapted to oxidative stress.

Relatively little is known about why odd-numbered fatty acids (OFAs) can be synthesized only by some plant species. We aimed at determining whether there is a relationship between the effects of Cd-induced oxidative stress on unsaturated fatty acids (USFAs) and their degradation products, especially OFAs. Plants with different ability to accumulate Cd - Noccaea praecox from Mezica, Slovenia (Me) and two ecotypes of Noccaea caerulescens from Ganges, France (Ga) and Redlschlag, Austria (Re) were cultivated in pot experiments. Only Me plants contained OFA 13:0, while all plants contained OFAs 15:0, 17:0 and 23:0 but in different proportions. Mutual correlations showed a significant effect of Cd contamination on the content of OFAs and USFAs in Me, a less pronounced effect in Re and the lowest one in Ga plants. The most significant correlation between the contents of USFAs and OFAs was also calculated for Me plants. The correlations between OFAs and USFAs indicate an active participation of OFA in FAs metabolism. Increased efficiency of utilization of the assimilated carbon via OFAs metabolism of Me plants in contrast to Re and Ga is also reflected in the increase of tolerance of Me plants to Cd toxicity in plant cells.

Utilization of biochar and activated carbon to reduce Cd, Pb and Zn phytoavailability and phytotoxicity for plants.

In the present study, the content of risk elements and content of free amino acids were studied in spinach (Spinacia oleracea L.) and mustard (Sinapis alba L.) subsequently grown on uncontaminated and contaminated soils (5 mg Cd/kg, 1000 mg Pb/kg and 400 mg Zn/kg) with the addition of activated carbon (from coconut shells) or biochar (derived from local wood residues planted for phytoextaction) in different seasons (spring, summer and autumn). The results showed that activated carbon and biochar increased biomass production on contaminated site. Application of amendments decreased Cd and Zn uptake by spinach plants. Mustard significantly increased Pb accumulation in the biomass as well in subsequently grown autumn spinach. Glutamic acid and glutamine were major free amino acids in leaves of all plants (15-34% and 3-45%) from total content. Application of activated carbon and biochar increased content of glutamic acid in all plants on uncontaminated and contaminated soils. Activated carbon and biochar treatments also induced an increase of aspartic acid in spinach plants. Biochar produced from biomass originated from phytoextraction technologies promoted higher spinach biomass yield comparing unamended control and showed a tendency to reduce accumulation of cadmium and zinc and thus it is promising soil amendment.

Responses to Cd Stress in Two Noccaea Species (Noccaea praecox and Noccaea caerulescens) Originating from Two Contaminated Sites in Mezica, Slovenia and Redlschlag, Austria.

The two Noccaea species-Noccaea praecox originating from Mezica, Slovenia (Me) (Pb, Zn, Cd pollution) and Noccaea caerulescens from Redlschlag, Austria (Re) (high levels of Ni, Cr, Mg)-were studied to compare Cd accumulation and tolerance. After 120 days of plant cultivation in Cd-contaminated soil (90 mg Cd kg(-1) soil), gas-exchange parameters (e.g. net photosynthetic rate, transpiration rate, stomatal conductance, and intercellular CO2 concentration), fatty acids, and selected macro- and microelements were determined in addition to N utilization by plants. The comparison between ecotypes showed that Cd stress resulted in similar changes in gas-exchange parameters. Contrasting responses of plants to Cd contamination were confirmed by the macro- and microelement contents and fatty acid and amino acid metabolism. Significantly higher accumulations of Cd and strong decreases in the levels of K, Ca, Na, and Fe were observed in the Me plants in contrast to the Re plants. The higher Re plant ability to take in some cations is a result of selective pressure due to contamination. Different ion uptake by plants affected the activities of metalloenzymes. Significant increases in the glutamic acid/proline ratio resulted from higher adaption of the Me in contrast to the Re plants.

Fatty acid profiles of ecotypes of hyperaccumulator Noccaea caerulescens growing under cadmium stress.

Changes in the fatty acid (FAs) composition in response to the extent of Cd contamination of soils (0, 30, 60 and 90 mg Cd kg(-1)) differed between ecotypes of Noccaea caerulescens originating from France - Ganges, Slovenia - Mezica and Austria - Redlschlag. Mezica ecotype accumulated more Cd in aboveground biomass compared to Ganges and Redlschlag ecotypes. Hyperaccumulators contained saturated fatty acids (SFAs) rarely occurring in plants, as are cerotic (26:0), montanic (28:0), melissic (30:0) acids, and unusual unsaturated fatty acids (USFAs), as are 16:2, 16:3, 20:2 and 20:3. Typical USFAs occurring in the family Brassicaceae, such as erucic, oleic and arachidonic acids, were missing in tested plants. Our results clearly indicate a relationship between Cd accumulation and the FAs composition. The content of SFAs decreased and the content of USFAs increased in aboveground biomass of Ganges and Mezica ecotypes with increasing Cd concentration. Opposite trend of FAs content was determined in Redlschlag ecotype. Linoleic (18:2n-6), α-linolenic (18:3n-3) and palmitic (16:0) acids were found in all ecotypes. The results observed in N. caerulescens ecotypes, showed that mainly Mezica ecotype has an efficient defense strategies which can be related on changes in FAs composition, mainly in VLCFAs synthesis. The most significant effect of ecotype on FAs composition was confirmed using multivariate analysis of variance.

Nitrogen metabolism and gas exchange parameters associated with zinc stress in tobacco expressing an ipt gene for cytokinin synthesis.

Increased endogenous plant cytokinin (CK) content through transformation with an isopentyl transferase (ipt) gene has been associated with improved plant stress tolerance. The impact of zinc (tested levels Zn1=250, Zn2=500, Zn3=750mgkg(-1)soil) on gas exchange parameters (net photosynthetic rate, transpiration rate, stomatal conductance, intercellular CO2 concentration) and nitrogen utilization by plants resulted in changes of free amino acid concentrations (glutamic acid, glutamine, asparagine, aspartate, glycine, serine, cystein) and differed for transformed and non-transformed tobacco plants. For pot experiments, tobacco plants (Nicotiana tabacum L., cv. Wisconsin 38) transformed with a construct consisting of SAG12 promoter fused with the ipt gene for cytokinin synthesis (SAG plants) and its wild type (WT plants as a control) were used. Physiological analyses confirmed that SAG plants had improved zinc tolerance compared with the WT plants. The enhanced Zn tolerance of SAG plants was associated with the maintenance of accumulation of amino acids and with lower declines of photosynthetic and transpiration rates. In comparison to WT plants, SAG plants exposed to the highest Zn concentration accumulated lower concentrations of asparagine, which is a major metabolic product during senescence.

The long-term effect of zinc soil contamination on selected free amino acids playing an important role in plant adaptation to stress and senescence.

Increased endogenous plant cytokinin (CK) content through transformation with an isopentyl transferase (ipt) gene has been associated with improved plant stress tolerance. The objective of this study is to determine amino acid changes associated with elevated CK production in ipt transgenic tobacco (Nicotiana tabacum L., cv. Wisconsin 38). Nontransformed (WT) and transformed tobacco plants with ipt gene controlled by senescence-activated promoter (SAG) were exposed to zinc soil contamination (tested levels Zn1=250, Zn2=500, Zn3=750 mg kg(-1) soil). The Zn effect on plant stress metabolism resulted in changes in levels of selected free amino acids playing an important role in adaptation to stress and plant senescence (alanine, leucine, proline, methionine and γ-aminobutyrate) and differed for transformed and nontransformed tobacco plants. Analyses of amino acids confirmed that SAG tobacco plants had improved zinc tolerance compared with the WT plants. The enhanced Zn tolerance of SAG plants was associated with the maintenance of accumulation of proline, methionine and γ-aminobutyrate. The concentrations of leucine and alanine did not show significant differences between plant lines.