Petr Soudek

Accumulation of heavy metals using Sorghum sp.

The essential requirement for the effective phytoremediation is selection of a plant species which should be metal tolerant, with high biomass production and known agronomic techniques. The above mentioned criteria are met by crop plant sorghum (Sorghum bicolor). The response of hydroponically grown S. bicolor plants to cadmium and zinc stress was followed. The impact of metal application on physiological parameters, including changes in chlorophylls contents and antioxidative enzymes activities, was followed during the stress progression. Cadmium and zinc were accumulated primarily in the roots of sorghum plants. However, elevation of metal concentrations in the media promoted their transfer to the shoots. Toxic effects of metals applied at lower concentrations were less serious in the shoots in comparison with their influence to the roots. When applied at higher concentrations, transfer of the metals into the leaves increased, causing growth reduction and leading to Chl loss and metal-induced chlorosis. Moreover, higher metal levels in the roots overcame the quenching capacity of peroxidase and glutathione transferase, which was associated with reduction of their activities. Fortification of antioxidant system by addition of glutathione significantly increased the accumulation of cadmium in the roots as well as in the shoots at the highest cadmium concentration applied.

Plant response to heavy metals and organic pollutants in cell culture and at whole plant level

In vitro cell culture experiments provide a convenient system to study basic biological processes, by which biochemical pathways, enzymatic activity and metabolites can be specifically studied. However, it is difficult to relate cell cultures, calli or even hydroponic experiments to the whole plant response to pollutant stress. In the field, plants are exposed to additional a-biotic and biotic factors, which complicate further plant response. Hence, we often see thatin vitro selected species perform poorly under soil and field conditions. Soil physical and chemical properties, plantmycorrhizal association and soil-microbial activity affect the process of contaminant degradation by plants and/or microorganisms, pointing to the importance of pot and field experiments.BackgroundIncreasing awareness in the last decade concerning environmental quality had prompted research into ‘green solutions’ for soil and water remediation, progressing from laboratoryin vitro experiments to pot and field trials.ObjectiveThis paper is a joint effort of a group of scientists in COST action 837. It represents experimental work and an overview on plant response to environmental stress fromin vitro tissue culture to whole plant experiments in soil.ResultsResults obtained fromin vitro plant tissue cultures and whole plant hydroponic experiments indicate the phytoremediation potential of different plant species and the biochemical mechanisms involved in plant tolerance. In pot experiments, several selected desert plant species, which accumulated heavy metal in hydroponic systems, succeeded in accumulating the heavy metal in soil conditions as well.Conclusions and RecommendationsIn vitro plant tissue cultures provide a useful experimental system for the study of the mechanisms involved in the detoxification of organic and heavy metal pollutants. However, whole plant experimental systems, as well as hydroponics followed by pot and field trials, are essential when determining plant potential to remediate polluted sites. Multidisciplinary research teams can therefore increase our knowledge and promote a practical application of phytoremediation.

Phytoremediation of Pharmaceuticals—Preliminary Study

Phytoremediation of selected pharmaceuticals (diclofenac, ibuprofen, and acetaminophen) using Armoracia rusticana and Linum usitatissimum cell cultures and by hydroponically cultivated Lupinus albus, Hordeum vulgaris, and Phragmites australis plants in laboratory conditions is described. During in vitro experiments, the best results for acetaminophen were achieved using Armoracia rusticana hairy root cultures, where 100% of the starting amount was removed from the media during eight days. Total removal of ibuprofen and diclofenac was achieved using a Linum usitatissimum suspension culture after one and six days, respectively. In the hydroponic arrangement, the best results were achieved for Lupinus, where acetaminophen was totally removed from media during two or four days in concentrations of 0.1 or 0.2 mM, respectively. The best effectiveness of ibuprofen removal (50% of starting amount) was found in case of Phragmites. Effectiveness of all tested plants for diclofenac removal was low. The best removal was achieved using Phragmites in the case of 0.2 mM concentration—67% of the starting amount and Hordeum for 0.1 mM starting concentration, 56%.

ACCUMULATION OF HEAVY METALS BY IN VITRO CULTURES OF PLANTS

The aim of the project is to study heavy metals accumulation by the selected plants in both laboratory and field conditions. Within the experiments the aspen (Populus tremula × tremuloides), sunflower (Helianthus annuus) and corn (Zea mays) plants were studied. The reasons for this selection were: a fast growth of these plants, an accumulation capacity and an ability to survive in different types of soils. The study was carried out on the aspen plantlets grown in vitro. The plants were exposed to the aqueous solutions having concentrations 0.1 mM, 0.5 mM of Pb2+ or Ni2+, respectively. The accumulation capacityfor aspen, was about 70% of Pb2+ originally present in the solution. The starting concentration of Pb2+ (0.5 mM) exhibited no negative impact on the growth. Besides in vitro expositions, a pilot-scale phytoremediation experiment was carried out at the polluted industrial area (Zn – 75000 mg/kg), (Pb – 16000 mg/kg), (Cr – 590 mg/kg), (Cd – 90 mg/kg) and (Cu – 1700 mg/kg).

Accumulation and Transformation of Sulfonated Aromatic Compounds by Rhubarb Cells (Rheum palmatum)

ABSTRACT Sulfonated aromatic compounds are released into the environment in large amounts: the main sources of these recalcitrant pollutants are anionic detergents, dyestuffs, and their byproducts, such as sulfonated anthraquinones. Different derivatives of anthraquinones occur naturally in several plant species such as rhubarb. Therefore, this plant could possess enzymes able to accept sulfonated anthraquinones as substrates. Previously, it has been shown that cultured cells isolated from Rheum palmatum are able to efficiently accumulate and transform different mono- and disulfonated anthraquinones. Because rhubarb is a hardy plant species, it may prove a promising candidate in developing new biological processes to decontaminate effluents containing recalcitrant xenobiotics. The ability of rhubarb cells to accumulate and transform other sulfonoaromatics was investigated in this study. Results obtained show that cultured rhubarb cells were able to efficiently accumulate 2-chloro-5-nitro-benzene sulfonate...

Uranium uptake by hydroponically cultivated crop plants.

Hydroponicaly cultivated plants were grown on medium containing uranium. The appropriate concentrations of uranium for the experiments were selected on the basis of a standard ecotoxicity test. The most sensitive plant species was determined to be Lactuca sativa with an EC(50) value about 0.1mM. Cucumis sativa represented the most resistant plant to uranium (EC(50)=0.71 mM). Therefore, we used the uranium in a concentration range from 0.1 to 1mM. Twenty different plant species were tested in hydroponic solution supplemented by 0.1mM or 0.5mM uranium concentration. The uranium accumulation of these plants varied from 0.16 mg/g DW to 0.011 mg/g DW. The highest uranium uptake was determined for Zea mays and the lowest for Arabidopsis thaliana. The amount of accumulated uranium was strongly influenced by uranium concentration in the cultivation medium. Autoradiography showed that uranium is mainly localized in the root system of the plants tested. Additional experiments demonstrated the possibility of influencing the uranium uptake from the cultivation medium by amendments. Tartaric acid was able to increase uranium uptake by Brassica oleracea and Sinapis alba up to 2.8 times or 1.9 times, respectively. Phosphate deficiency increased uranium uptake up to 4.5 times or 3.9 times, respectively, by Brassica oleracea and S. alba. In the case of deficiency of iron or presence of cadmium ions we did not find any increase in uranium accumulation.

Study of soil-plant transfer of 226Ra under greenhouse conditions.

A soil-plant transfer study was performed using soil from a former uranium ore processing factory in South Bohemia. We present the results from greenhouse experiments which include estimates of the time required for phytoremediation. The accumulation of (226)Ra by different plant species from a mixture of garden soil and contaminated substrate was extremely variable, ranging from 0.03 to 2.20 Bq (226)Ra/g DW. We found differences in accumulation of (226)Ra between plants from the same genus and between cultivars of the same plant species. The results of (226)Ra accumulation showed a linear relation between concentration of (226)Ra in plants and concentration of (226)Ra in soil mixtures. On the basis of these results we estimated the time required for phytoremediation, but this appears to be too long for practical purposes.

Optimalization of the peroxidase production by tissue cultures of horseradish in vitro

Tissue cultures of Armoracia rusticana L., both transformed with Agrobacterium rhizogenes and nontransformed, were screened for peroxidase activity. Most of the derived and tested strains exhibited 20 times higher activity [from 99 to 723 U g−1(d.m.)] than the root of the intact plant [(30 U g−1 (d.m.)]. The highest peroxidase activity was found in tumour culture growing on the medium without growth regulators. The influence of the addition of sugars and heavy metal ions in the medium on peroxidase production was tested. Increase in peroxidase activity was observed after cultivation of horseradish culture with cadmium, cobalt, nickel or lead ions.

Preliminary study of phytoremediation of brownfield soil contaminated by PAHs

Our project was aimed at improving a brownfield in the city of Kladno, where an old steel producing facility used to be in operation. Ecological risk is mainly caused by the processing of co-products during coal production (tars, oils). Knowledge of toxicology and environmental aspects can help us protect human health and the environment. Primarily, we focused on soil sampling and identification of pollutants. Results showed that organic contamination on the site is very high. Average concentration of total petroleum carbon in the soil was about 13g/kg DW, which is much more than the maximum allowed concentration. For selection of suitable plant species for phytoremediation at the site, experiments were conducted in a greenhouse. Biomass growth, root morphology, and pigment content in the leaves of Brassica napus var. Opus-C1 and Sorghum×drummondii var. Honey Graze BMR plants were studied. Plant analysis confirmed that polyaromatic hydrocarbons (PAHs) accumulated in the shoots of both plant species. B. napus plants grown on Poldi soil in a greenhouse were able to survive the toxicity of PAHs in soil, and their ability to accumulate PAHs from soil was evident. However, more studies are needed to decide if the plants are usable for phytoremediation of this brownfield.

Improving crop tolerance to heavy metal stress by polyamine application.

Many areas have been heavily contaminated by heavy metals from industry and are not suitable for food production. The consumption of contaminated foods represents a health risk in humans, although some heavy metals are essential at low concentrations. Increasing the concentrations of essential elements in foods is one goal to improve nutrition. The aim of this study was to increase the accumulation of heavy metals in plant foods by the external application of putrescine. The levels of cadmium, zinc and iron were measured in different vegetables grown in hydroponic medium supplemented with heavy metals and compared with those grown in a reference medium. The estimated daily intake, based on the average daily consumption for various vegetable types, and the influence of polyamines on metal uptake were calculated.