Adaptation of Arabidopsis halleri to extreme metal pollution through limited metal accumulation involves changes in cell wall composition and metal homeostasis.

Metallophytes constitute powerful models for the study of metal homeostasis, adaptation to extreme environments and the evolution of naturally selected traits. Arabidopsis halleri is a pseudometallophyte which shows constitutive zinc/cadmium (Zn/Cd) tolerance and Zn hyperaccumulation but high intraspecific variability in Cd accumulation. To examine the molecular basis of the variation in metal tolerance and accumulation, ionome, transcriptome and cell wall glycan array profiles were compared in two genetically close A. halleri populations from metalliferous and nonmetalliferous sites in Northern Italy. The metallicolous population displayed increased tolerance to and reduced hyperaccumulation of Zn, and limited accumulation of Cd, as well as altered metal homeostasis, compared to the nonmetallicolous population. This correlated well with the differential expression of transporter genes involved in trace metal entry and in Cd/Zn vacuolar sequestration in roots. Many cell wall-related genes were also more highly expressed in roots of the metallicolous population. Glycan array and histological staining analyses demonstrated that there were major differences between the two populations in terms of the accumulation of specific root pectin and hemicellulose epitopes. Our results support the idea that both specific cell wall components and regulation of transporter genes play a role in limiting accumulation of metals in A. halleri at contaminated sites.

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.

Hormesis in Plants: The Role of Oxidative Stress, Auxins and Photosynthesis in Corn Treated with Cd or Pb.

Hormesis, which describes the stimulatory effect of low doses of toxic substances on growth, is a well-known phenomenon in the plant and animal kingdoms. However, the mechanisms that are involved in this phenomenon are still poorly understood. We performed preliminary studies on corn coleoptile sections, which showed a positive correlation between the stimulation of growth by Cd or Pb and an increase in the auxin and H2O2 content in the coleoptile sections. Subsequently, we grew corn seedlings in hydroponic culture and tested a wide range of Cd or Pb concentrations in order to determine hormetic growth stimulation. In these seedlings the gas exchange and the chlorophyll a fluorescence, as well as the content of chlorophyll, flavonol, auxin and hydrogen peroxide, were measured. We found that during the hormetic stimulation of growth, the response of the photosynthetic apparatus to Cd and Pb differed significantly. While the application of Cd mostly caused a decrease in various photosynthetic parameters, the application of Pb stimulated some of them. Nevertheless, we discovered that the common features of the hormetic stimulation of shoot growth by heavy metals are an increase in the auxin and flavonol content and the maintenance of hydrogen peroxide at the same level as the control plants.

Contrasting cadmium resistance strategies in two metallicolous populations of Arabidopsis halleri.

While cadmium (Cd) tolerance is a constitutive trait in the Arabidopsis halleri species, Cd accumulation is highly variable. Recent adaptation to anthropogenic metal stress has occurred independently within the genetic units of A. halleri and the evolution of different mechanisms involved in Cd tolerance and accumulation has been suggested. To gain a better understanding of the mechanisms underlying Cd tolerance and accumulation in A. halleri, ionomic inductively coupled plasma mass spectrometry (ICP-MS), transcriptomic (RNA sequencing) and metabolomic (high-performance liquid chromatography-mass spectrometry) profiles were analysed in two A. halleri metallicolous populations from different genetic units (PL22 from Poland and I16 from Italy). The PL22 and I16 populations were both hypertolerant to Cd, but PL22 hyperaccumulated Cd while I16 behaved as an excluder both in situ and when grown hydroponically. The observed hyperaccumulator vs excluder behaviours were paralleled by large differences in the expression profiles of transporter genes. Flavonoid-related transcripts and metabolites were strikingly more abundant in PL22 than in I16 shoots. The role of novel A. halleri candidate genes possibly involved in Cd hyperaccumulation or exclusion was supported by the study of corresponding A. thaliana knockout mutants. Taken together, our results are suggestive of the evolution of divergent strategies for Cd uptake, transport and detoxification in different genetic units of A. halleri.

Photosynthetic Efficiency as Bioindicator of Environmental Pressure in A. halleri.

In earlier ecophysiological studies that were conducted on Arabidopsis halleri plants, scientists focused on the mechanisms of Cd and Zn hyperaccumulation but did not take into consideration the environmental factors that can significantly affect the physiological responses of plants in situ. In this study, we investigated A. halleri that was growing on two nonmetalliferous and three metalliferous sites, which were characterized by different environmental conditions. We compared these populations in order to find differences within the metallicolous and nonmetallicolous groups that have not yet been investigated. The concentrations of several elements in the plant and soil samples also were investigated. To our knowledge, the concentration and fluorescence of chlorophyll were measured for A. halleri in situ for the first time. Our study confirmed the hyperaccumulation of Cd and Zn for each metallicolous population. For the metallicolous populations, the inhibition of parameters that describe the efficiency of the photosynthetic apparatus with increasing accumulations of heavy metals in the shoots also was observed. It was found that the nonmetallicolous plant populations from the summit of Ciemniak Mountain had larger antenna dimensions and chlorophyll content but a lower percentage of active reaction centers. To our knowledge, in this study, the internal high physiological diversity within the populations that inhabit metalliferous and nonmetalliferous sites is presented for the first time.

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.

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.

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.

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.

Toxic Effects of Cd and Zn on the Photosynthetic Apparatus of the Arabidopsis halleri and Arabidopsis arenosa Pseudo-Metallophytes.

Hyperaccumulation and hypertolerance of Trace Metal Elements (TME) like Cd and Zn are highly variable in pseudo-metallophytes species. In this study we compared the impact of high Cd or Zn concentration on the photosynthetic apparatus of the Arabidopsis arenosa and Arabidopsis halleri pseudo-metallophytes growing on the same contaminated site in Piekary Slaskie in southern Poland. Plants were grown in hydroponic culture for 6 weeks, and then treated with 1.0 mM Cd or 5.0 mM Zn for 5 days. Chlorophyll a fluorescence and pigment content were measured after 0, 1, 2, 3, 4, and 5 days in plants grown in control and exposed to Cd or Zn treatments. Moreover, the effect of TME excess on the level of oxidative stress and gas-exchange parameters were investigated. In both plant species, exposure to high Cd or Zn induced a decrease in chlorophyll and an increase in anthocyanin contents in leaves compared to the control condition. After 5 days Cd treatment, energy absorbance, trapped energy flux and the percentage of active reaction centers decreased in both species. However, the dissipated energy flux in the leaves of A. arenosa was smaller than in A. halleri. Zn treatment had more toxic effect than Cd on electron transport in A. halleri compared with A. arenosa. A. arenosa plants treated with Zn excess did not react as strongly as in the Cd treatment and a decrease only in electron transport flux and percentage of active reaction centers compared with control was observed. The two species showed contrasting Cd and Zn accumulation. Cd concentration was almost 3-fold higher in A. arenosa leaves than in A. halleri. On the opposite, A. halleri leaves contained 3-fold higher Zn concentration than A. arenosa. In short, our results showed that the two Arabidopsis metallicolous populations are resistant to high Cd or Zn concentration, however, the photosynthetic apparatus responded differently to the toxic effects.

Influence of short-term macronutrient deprivation in maize on photosynthetic characteristics, transpiration and pigment content.

The aim of the research was to compare the impact of short-term deprivation of selected macronutrients (Ca, K, Mg and P) on the photosynthetic characteristics, transpiration and pigment content in maize. The strongest inhibition of photosynthesis was caused by a deprivation of Mg, which was visible as a decrease in the photosynthetic and transpiration rates, stomatal conductance, photosystem II (PSII) performance, chlorophyll and flavonol content with a simultaneously increased content of anthocyanins. In the K-deprived plants, a decrease in the photosynthetic rate was observed. However, the transpiration rate and stomatal conductance did not differ significantly compared with the control. In the K-deprived plants, a decrease in chlorophyll and an increase in the anthocyanin content were also observed. We showed that Ca starvation resulted in a decrease in the photosynthetic and transpiration rates, stomatal conductance and PSII performance, while the pigment content was not significantly different compared with the control. In the case of P-deprived plants, we observed a decrease in the photosynthetic and transpiration rates. Interestingly, the inhibition of stomatal conductance was the strongest in the P-deprived plants compared with all of the investigated elements. However, the performance of PSII was not significantly affected by P starvation compared with the control. Our results present for the first time a comprehensive analysis of the effect of short-term macronutrient deprivation on photosynthesis and transpiration in maize plants.

How autochthonous microorganisms influence physiological status of Zea mays L. cultivated on heavy metal contaminated soils?

The aim of this study was to investigate the effect of autochthonous microorganisms present in soil collected from heavy metal (HM) uncontaminated (Pb ≈ 59 mg kg-1, Cd ≈ 0.4 mg kg-1, Zn ≈ 191 mg kg-1), moderately (Pb ≈ 343 mg kg-1, Cd ≈ 12 mg kg-1, Zn ≈ 1876 mg kg-1), and highly (Pb ≈ 1586 mg kg-1, Cd ≈ 57 mg kg-1, Zn ≈ 3280 mg kg-1) contaminated sites on Zea mays elemental composition, physiological status, and growth parameters. For this purpose, half of the collected soil was sterilized and soil characterization was performed. After 45 days of cultivation, the presence of HM in the soil negatively affected photosynthesis and transpiration rates, relative chlorophyll content, anthocyanins index, chlorophyll fluorescence parameters, and content of oxidative stress products (H2O2 and Malondialdehyde) of Zea mays, while soil sterilization had a positive effect on those parameters. Average percentage of colonization of root segments by arbuscular mycorrhiza fungi decreased with an increase of HM contamination in the soil. The increase in shoot concentration of HMs, particularly Cd and Zn, was a result of contaminated soils sterilization. Aboveground biomass of maize cultivated on sterilized soil was 3-fold, 1.5-fold, and 1.5-fold higher for uncontaminated, moderately contaminated and highly contaminated soils respectively when compared to nonsterilized soils. Contrary to our expectation, autochthonous microflora did not improve plant growth and photosynthetic performance; in fact, they had a negative effect on those processes although they did reduce concentration of HMs in the shoots grown on contaminated soils.

Biominerals and waxes of Calamagrostis epigejos and Phragmites australis leaves from post-industrial habitats.

Vascular plants are able to conduct biomineralization processes and collect synthesized compounds in their internal tissues or to deposit them on their epidermal surfaces. This mechanism protects the plant from fluctuations of nutrient levels caused by different levels of supply and demand for them. The biominerals reflect both the metabolic characteristics of a vascular plant species and the environmental conditions of the plant habitat. The SEM/EDX method was used to examine the surface and cross-sections of the Calamagrostis epigejos and Phragmites australis leaves from post-industrial habitats (coal and zinc spoil heaps). The results from this study have showed the presence of mineral objects on the surfaces of leaves of both grass species. The calcium oxalate crystals, amorphous calcium carbonate spheres, and different silica forms were also found in the inner tissues. The high variety of mineral forms in the individual plants of both species was shown. The waxes observed on the leaves of the studied plants might be the initializing factor for the crystalline forms and structures that are present. For the first time, wide range of crystal forms is presented for C. epigejos. The leaf samples of P. australis from the post-industrial areas showed an increased amount of mineral forms with the presence of sulfur.

Effect of chemophytostabilization practices on arbuscular mycorrhiza colonization of Deschampsia cespitosa ecotype Warynski at different soil depths.

The effects of chemophytostabilization practices on arbuscular mycorrhiza (AM) of Deschampsia cespitosa roots at different depths in soils highly contaminated with heavy metals were studied in field trials. Mycorrhizal parameters, including frequency of mycorrhization, intensity of root cortex colonization and arbuscule abundance were studied. Correlations between concentration of bioavailable Cd, Zn, Pb and Cu in soil and mycorrhizal parameters were estimated. An increase in AM colonization with increasing soil depth was observed in soils with spontaneously growing D. cespitosa. A positive effect of chemophytostabilization amendments (calcium phosphate, lignite) on AM colonization was found in the soil layers to which the amendments were applied. Negative correlation coefficients between mycorrhizal parameters and concentration of bioavailable Cd and Zn in soil were obtained. Our results demonstrated that chemophytostabilization practices enhance AM colonization in D. cespitosa roots, even in soils fertilized with high rates of phosphorus.

Relationships between soil parameters and physiological status of Miscanthus x giganteus cultivated on soil contaminated with trace elements under NPK fertilisation vs. microbial inoculation.

Crop growth and development can be influenced by a range of parameters, soil health, cultivation and nutrient status all play a major role. Nutrient status of plants can be enhanced both through chemical fertiliser additions (e.g. N, P, K supplementation) or microbial fixation and mobilisation of naturally occurring nutrients. With current EU priorities discouraging the production of biomass on high quality soils there is a need to investigate the potential of more marginal soils to produce these feedstocks and the impacts of soil amendments on crop yields within them. This study investigated the potential for Miscanthus x giganteus to be grown in trace element (TE)-contaminated soils, ideally offering a mechanism to (phyto)manage these contaminated lands. Comprehensive surveys are needed to understand plant-soil interactions under these conditions. Here we studied the impacts of two fertiliser treatments on soil physico-chemical properties under Miscanthus x giganteus cultivated on Pb, Cd and Zn contaminated arable land. Results covered a range of parameters, including soil rhizosphere activity, arbuscular mycorrhization (AM), as well as plant physiological parameters associated with photosynthesis, TE leaf concentrations and growth performance. Fertilization increased growth and gas exchange capacity, enhanced rhizosphere microbial activity and increased Zn, Mg and N leaf concentration. Fertilization reduced root colonisation by AMF and caused higher chlorophyll concentration in plant leaves. Microbial inoculation seems to be a promising alternative for chemical fertilizers, especially due to an insignificant influence on the mobility of toxic trace elements (particularly Cd and Zn).

Interactive effects of temperature and heavy metals (Cd, Pb) on the elongation growth in maize coleoptiles.

The effect of Cd and Pb on endogenous and IAA-induced elongation growth and medium pH of maize coleoptile segments incubated at 20, 25 and 30 °C was studied. It was found that the elongation of coleoptile segments and proton extrusion increased with the temperature and reached its maximum at 30 °C. For Cd, the maximal inhibition of endogenous and IAA-induced growth as well as medium acidification of coleoptile segments was observed at 25 °C. Meanwhile, Pb, irrespective of the temperature, diminished the growth of the segments by ca. 20%, increasing the acidification of the incubation medium. It was also found that in contrast to Cd, Pb accumulation in maize coleoptile segments did not correlate with temperature. The results suggest that the toxic effect of Cd on elongation growth of coleoptile segments is connected with the decrease of the PM H(+)-ATPase activity and probably with Cd-induced high acivity of IAA oxidase, whereas the effect of Pb did not depend on activity of any of the enzymes.

Remediation aspect of microbial changes of plant rhizosphere in mercury contaminated soil.

Phytoremediation, an approach that uses plants to remediate contaminated soil through degradation, stabilization or accumulation, may provide an efficient solution to some mercury contamination problems. This paper presents growth chamber experiments that tested the ability of plant species to stabilize mercury in soil. Several indigenous herbaceous species and Salix viminalis were grown in soil collected from a mercury-contaminated site in southern Poland. The uptake and distribution of mercury by these plants were investigated, and the growth and vitality of the plants through a part of one vegetative cycle were assessed. The highest concentrations of mercury were found at the roots, but translocation to the aerial part also occurred. Most of the plant species tested displayed good growth on mercury contaminated soil and sustained a rich microbial population in the rhizosphere. The microbial populations of root-free soil and rhizosphere soil from all species were also examined. An inverse correlation between the number of sulfur amino acid decomposing bacteria and root mercury content was observed. These results indicate the potential for using some species of plants to treat mercury contaminated soil through stabilization rather than extraction. The present investigation proposes a practical cost-effective temporary solution for phytostabilization of soil with moderate mercury contamination as well as the basis for plant selection.