The traditional utilization, biological activity and chemical composition of edible fern species.

ETHNOPHARMACOLOGICAL RELEVANCE: Ferns form an important part of the human diet. Young fern fiddleheads are mostly consumed as vegetables, while the rhizomes are often extracted for starch. These edible ferns are also often employed in traditional medicine, where all parts of the plant are used, mostly to prepare extracts. These extracts are applied either externally as lotions and baths or internally as potions, decoctions and teas. Ailments traditionally treated with ferns include coughs, colds, fevers, pain, burns and wounds, asthma, rheumatism, diarrhoea, or skin diseases (eczema, rashes, itching, leprosy). AIM OF THE REVIEW: This review aims to compile the worldwide knowledge on the traditional medicinal uses of edible fern species correlating to reported biological activities and isolated bioactive compounds. MATERIALS AND METHODS: The articles and books published on edible fern species were searched through the online databases Web of Science, Pubmed and Google Scholar, with critical evaluation of the hits. The time period up to the end of 2022 was included. RESULTS: First, the edible fern species were identified based on the literature data. A total of 90 fern species were identified that are eaten around the world and are also used in traditional medicine. Ailments treated are often associated with inflammation or bacterial infection. However, only the most common and well-known fern species, were investigated for their biological activity. The most studied species are Blechnum orientale L., Cibotium barometz (L.) J. Sm., Diplazium esculentum (Retz.) Sw., Marsilea minuta L., Osmunda japonica Thunb., Polypodium vulgare L., and Stenochlaena palustris (Burm.) Bedd. Most of the fern extracts have been studied for their antioxidant, anti-inflammatory and antimicrobial activities. Not surprisingly, antioxidant capacity has been the most studied, with results reported for 28 edible fern species. Ferns have been found to be very rich sources of flavonoids, polyphenols, polyunsaturated fatty acids, carotenoids, terpenoids and steroids and most of these compounds are remarkable free radical scavengers responsible for the outstanding antioxidant capacity of fern extracts. As far as clinical trials are concerned, extracts from only three edible fern species have been evaluated. CONCLUSIONS: The extracts of edible fern species exert antioxidant anti-inflammatory and related biological activities, which is consistent with their traditional medicinal use in the treatment of wounds, burns, colds, coughs, skin diseases and intestinal diseases. However, studies to prove pharmacological activities are scarce, and require chemical-biological standardization. Furthermore, correct botanical classification needs to be included in publications to simplify data acquisition. Finally, more in-depth phytochemical studies, allowing the linking of traditional use to pharmacological relevance are needed to be done in a standardized way.

Ecotoxicity Study of Additives Composed of Zinc and Boron.

The high use of additives containing zinc borate and their limited solubility in water both lead to its persistence and accumulation in biological systems. On the other hand, soluble forms of boron are easily available to plant roots and are taken up by plants. There are no ecotoxicological data available for zinc borate, the industrial utilization of which is widespread. Therefore, the potential toxicity of zinc borate and its dissociated compounds was evaluated. Based on two different ecotoxicology tests, their effect on plant growth was studied. Firstly, the impact on Lemna minor growth was investigated, including the effect on pigment content. Secondly, the inhibition of the root growth of higher plant species Sinapis alba (mustard), Lactuca sativa (lettuce) and Trifolium pretense (clover) was measured. The growth inhibition test on L. minor was more complex and sensitive compared to the plant seed germination test. Already low concentrations (10 mg/L) of ZnO, B2O3 and Zn3BO6 led to a decrease in frond growth and to an inhibition of the conversion of chlorophyll a to chlorophyll b. These results suggested that the stress caused by these additives caused damage to the photosynthetic apparatus. The highest inhibition of frond growth was detected in fronds treated with B2O3 (92-100%). In ZnO and Zn3BO6, the inhibition of frond growth was between 38 and 77%, with Zn3BO6 being slightly more toxic. In the seed germination test, the most sensitive species was lettuce, the growth of which was inhibited by 57, 83 and 53% in ZnO, B2O3 and Zn3BO6 treatments, respectively. However, the inhibitory effect on each plant was different. In lettuce and clover, the seed germination and root elongation decreased with increasing element concentrations. In contrast, in mustard, low concentrations of ZnO and Zn3BO6 supported the growth of roots. For that reason, more complex tests are essential to evaluate the additive toxicity in the environment.

Biochar in manure can suppress water stress of sugar beet (Beta vulgaris) and increase sucrose content in tubers.

Increased soil drought events threaten the yields of sugar beet (Beta vulgaris L.) and other staples of arable production in central Europe. In this study we evaluated soil moisture and nutrients as impacted by a two and five % (wt) addition of biochar, manure and their blend to a loamy-sand Regosol. Cyclical soil drought was achieved by the controlled reduction of watering by 75% in pot experiments. Ongoing soil moisture and nutrient measurements were taken, and physiological parameters of sugar beet plants were analysed three weeks after the induced drought. At the end of the experiment (16 weeks) plants were harvested and their mass assessed, as well as their nutrient, pigment and sugar contents. In contrast to the addition of manure, soil volumetric water contents were two to three times greater after biochar amendment, compared to the control soil. Porewater analysis revealed that nutrient leaching (e.g., NO3-, K+) from manure addition to soil was reduced when biochar was blended in (by ≤86% compared to manure alone). Crop analysis showed that leaf gas exchanges were moderated during drought following soil amendment, and leaf and tuber yields were increased furthest when combined biochar-manure blends were applied (> 2-times compared to the control). Perhaps most importantly, the advantageous soil conditions induced by the combined biochar and manure addition also resulted in significantly increased sugar contents in plants (2.4-times) pointing to immediate practical applications of these results in the field.

Physiological and Transcriptomic Response of Grey Poplar (Populus ×canescens Aiton Sm.) to Cadmium Stress.

(1) Background: Populus ×canescens (Aiton) Sm. is a fast-growing woody plant belonging to the family Salicaceae. Two poplar genotypes characterized by unique phenotypic traits (TP11 and TP20) were chosen to be characterized and tested for a physiological and transcriptomic response to Cd stress. (2) Methods: A comparative analysis of the effects of exposure to high cadmium (Cd) concentrations (10 µM and 100 µM) of TP11 and TP20 was performed. (3) Results: Neither of the tested Cd concentration negatively affected plant growth; however, the chlorophyll content significantly decreased. The potassium (K) content was higher in the shoots than in the roots. The magnesium concentrations were only slightly affected by Cd treatment. The zinc content in the shoots of TP20 was lower than that in the shoots of TP11. Cd accumulation was higher in the roots than in the shoots. After 10 days of exposure, 10 µM Cd resulted in comparable amounts of Cd in the roots and shoots of TP20. The most significant change in transcript amount was observed in endochitinase 2, 12-oxophytodienoate reductase 1 and phi classglutathione S-transferase. (4) Conclusions: Our study provided new insights for effective assessing the ability of different poplar genotypes to tolerate Cd stress and underlying Cd tolerance.

Multi-Component Antioxidative System and Robust Carbohydrate Status, the Essence of Plant Arsenic Tolerance.

Arsenic (As) contaminates the food chain and decreases agricultural production through impairing plants, particularly due to oxidative stress. To better understand the As tolerance mechanisms, two contrasting tobacco genotypes: As-sensitive Nicotiana sylvestris and As-tolerant N.tabacum, cv. 'Wisconsin' were analyzed. The most meaningful differences were found in the carbohydrate status, neglected so far in the As context. In the tolerant genotype, contrary to the sensitive one, net photosynthesis rates and saccharide levels were unaffected by As exposure. Importantly, the total antioxidant capacity was far stronger in the As-tolerant genotype, based on higher antioxidants levels (e.g., phenolics, ascorbate, glutathione) and activities and/or appropriate localizations of antioxidative enzymes, manifested as reverse root/shoot activities in the selected genotypes. Accordingly, malondialdehyde levels, a lipid peroxidation marker, increased only in sensitive tobacco, indicating efficient membrane protection in As-tolerant species. We bring new evidence of the orchestrated action of a broad spectrum of both antioxidant enzymes and molecules essential for As stress coping. For the first time, we propose robust carbohydrate metabolism based on undisturbed photosynthesis to be crucial not only for subsidizing C and energy for defense but also for participating in direct reactive oxygen species (ROS) quenching. The collected data and suggestions can serve as a basis for the selection of plant As phytoremediators or for targeted breeding of tolerant crops.

Application of co-composted biochar significantly improved plant-growth relevant physical/chemical properties of a metal contaminated soil.

A woody-biochar was added to waste biomass during a composting process. The resulting compost-char was amended to a metal contaminated soil and two plant species, L. perenne and E. sativa, were grown in a pot experiment to determine 1) plant survival and stress factors, 2) uptake of metals to plants and, 3) chemical characteristics of sampled soils and pore waters. Compost supplemented with biochar after the composting process were also tested, as well as a commercially available compost, for comparison. Co-composting with biochar hastened the composting process, resulting in a composite material of reduced odour, increased maturity, circum-neutral pH and increased moisture retention than compost (increase by 3% of easily removable water content). When amended to the soil, CaCl2 extractable and pore water metals s were reduced by all compost treatments with little influence of biochar addition at any tested dose. Plant growth success was promoted furthest by the addition of co-composted biochar to the test soil, especially in the case of E. sativa. For both tested plant species significant reductions in plant metal concentrations (e.g. 8-times for Zn) were achieved, against the control soil, by compost, regardless of biochar addition. The results of this study demonstrate that the addition of biochar into the composting process can hasten the stability of the resulting compost-char, with more favourable characteristics as a soil amendment/improver than compost alone. This appears achievable whilst also maintaining the provision of available nutrients to soils and the reduction of metal mobility, and improved conditions for plant establishment.

Triazolide strigolactone mimics as potent selective germinators of parasitic plant Phelipanche ramosa.

BACKGROUND: Strigolactones are a unique class of plant metabolites which serve as a rhizosphere signal for parasitic plants and evocate their seed germination. The expansion of these parasitic weeds in the food crop fields urgently calls for their increased control and depletion. Simple strigolactone analogues able to stimulate seed germination of these parasitic plants may represent an efficient control measure through the induction of suicidal germination. RESULTS: Triazolide-type strigolactone mimics were easily synthesized in three steps from commercially available materials. These derivatives induced effectively seed germination of Phelipanche ramosa with EC50 as low as 5.2 × 10-10 M. These mimics did not induce seed germination of Striga hermonthica even at high concentration (≥1 × 10-5 M). CONCLUSIONS: Simple and stable strigolactone mimics with selective activity against Phelipanche ramosa were synthesized. © 2019 Society of Chemical Industry.

Resorcinol-Type Strigolactone Mimics as Potent Germinators of the Parasitic Plants Striga hermonthica and Phelipanche ramosa.

Strigolactones are a particular class of plant metabolites with diverse biological functions starting from the stimulation of parasitic seed germination to phytohormonal activity. The expansion of parasitic weeds in the fields of developing countries is threatening the food supply and calls for simple procedures to combat these weeds. Strigolactone analogues represent a promising approach for such control through suicidal germination, i.e., parasitic seed germination without the presence of the host causing parasite death. In the present work, the synthesis of resorcinol-type strigolactone mimics related to debranones is reported. These compounds were highly stable even at alkaline pH levels and able to induce seed germination of parasitic plants Striga hermonthica and Phelipanche ramosa at low concentrations, EC50 ≈ 2 × 10-7 M ( Striga) and EC50 ≈ 2 × 10-9 M ( Phelipanche). On the other hand, the mimics had no significant effect on root architecture of Arabidopsis plants, suggesting a selective activity for parasitic seed germination, making them a primary target as suicidal germinators.

Hormonal dynamics during salt stress responses of salt-sensitive Arabidopsis thaliana and salt-tolerant Thellungiella salsuginea.

Salt stress responses in salt-sensitive Arabidopsis thaliana (2-150mM NaCl) and the closely related salt-tolerant Thellungiella salsuginea (Eutrema halophila, 150-350mM NaCl) were compared to identify hormonal and transcriptomic changes associated with enhanced stress tolerance. Phytohormone levels, expression of selected genes, membrane stability, and Na+ and K+ concentrations were measured in shoot apices, leaves, and roots. Thellungiella exhibited higher salt stress tolerance associated with elevated basal levels of abscisic acid and jasmonic acid, and lower levels of active cytokinins (excluding cis-zeatin) in shoot apices. Analysis of the dynamics of the early salt stress response (15min to 24h) revealed that the halophyte response was faster and stronger. Very mild stress, in our hydropony arrangement 2-25mM NaCl, affected the transcription of genes involved in cytokinin metabolism (AtIPTs, AtCKXs). Mild stress induced in Arabidopsis (50mM) stress responses only in shoot apices, while in Thellungiella (150mM) across the whole plant. Arabidopsis exhibited in hydropony evidence of severe stress above 75mM NaCl and died in 150mM, whereas the halophyte only became severely stressed above 225mM. The responses of individual phytohormones (cytokinins, auxin, abscisic acid, jasmonic acid, salicylic acid and their metabolites) to salinity are discussed.

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.

Triazolide Strigolactone Mimics Influence Root Development in Arabidopsis.

Strigolactones are the most recently recognized class of phytohormones, which are also known to establish plant symbiosis with arbuscular mycorhizal fungi or induce germination of parasitic plants. Their relatively complex structures and low stability urgently calls for simple derivatives with maintained biological function. We have prepared a series of triazolide strigolactone mimics and studied their ability to affect root development of Arabidopsis thaliana. The strigolactone mimics significantly induced root elongation and lateral root formation while resembling the effect of the reference compound GR24.

Thorium impact on tobacco root transcriptome.

Thorium is natural actinide metal with potential use in nuclear energetics. Contamination by thorium, originated from mining activities or spills, represents environmental risk due to its radioactivity and chemical toxicity. A promising approach for cleaning of contaminated areas is phytoremediation, which need to be based, however, on detail understanding of the thorium effects on plants. In this study we investigated transcriptomic response of tobacco roots exposed to 200µM thorium for one week. Thorium application resulted in up-regulation of 152 and down-regulation of 100 genes (p-value <0.01, fold change ≥2). The stimulated genes were involved in components of jasmonic acid and salicylic acid signaling pathways and various abiotic (e.g. oxidative stress) and biotic stress (e.g. pathogens, wounding) responsive genes. Further, up-regulation of phosphate starvation genes and down-regulation of genes involved in phytic acid biosynthesis indicated that thorium disturbed phosphate uptake or signaling. Also expression of iron responsive genes was influenced. Negative regulation of several aquaporins indicated disturbance of water homeostasis. Genes potentially involved in thorium transport could be zinc-induced facilitator ZIF2, plant cadmium resistance PCR2, and ABC transporter ABCG40. This study provides the first insight at the processes in plants exposed to thorium.

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.

Characterization of contaminants and evaluation of the suitability for land application of maize and sludge biochars.

Prior to the application of biochar as an agricultural improver, attention should be paid to the potential introduction of toxicants and resulting unintended impacts on the environment. In the present study, the concentrations of polycyclic aromatic hydrocarbons (PAHs), heavy metals, and mineral elements were determined in maize and sludge biochars produced at 100 °C increments between 200 and 700 °C. The concentration ranges of total PAHs were 358-5,136 µg kg(-1) in maize biochars and 179-70,385 µg kg(-1) in sludge biochars. The total heavy metals were detected at the following concentrations (mg kg(-1)): Cu, 20.4-56.7; Zn, 59.7-133; Pb, 1.44-3.50; Cd, <0.014; Cr, 8.08-21.4; Ni, 4.38-9.82 in maize biochars and Cu, 149-202; Zn, 735-986; Pb, 54.7-74.2; Cd, 1.06-1.38; Cr, 180-247; Ni, 41.1-56.1 in sludge biochars. The total concentrations of PAHs and heavy metals in all maize biochars and most sludge biochars were below the control standards of sludge for agricultural use in China, the USA, and Europe. The leachable Mn concentrations in sludge biochars produced at below 500 °C exceeded the groundwater or drinking water standards of these countries. Overall, all the maize biochars were acceptable for land application, but sludge biochars generated at temperatures between 200 and 500 °C were unsuitable for application as soil amendments due to their potential adverse effects on soil and groundwater quality.

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.

Composition of hydroponic medium affects thorium uptake by tobacco plants.

The ability of thorium uptake as well as responses to heavy metal stress were tested in tobacco cultivar La Burley 21. Thorium was accumulated preferentially in the root system. The presence of citric, tartaric and oxalic acids in hydroponic medium increased thorium accumulation in all plant organs. On the other hand, the addition of diamines and polyamines, the important antioxidants in plants, resulted in decrease of thorium accumulation, especially in the root system. Negative correlation was found between putrescine concentration and thorium accumulation. Nevertheless, the most important factor influencing the accumulation of thorium was the absence of phosphate ions in a hydroponic medium that caused more than 10-fold increase of thorium uptake in all plant parts. Accumulation and distribution of thorium was followed in six cultivars and 14 selected transformants. Cultivar La Barley 21 represented an average between the tested genotypes, having a very good distribution ratio between roots, stems and leaves.

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.

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%.

Effect of heavy metals on inhibition of root elongation in 23 cultivars of flax (Linum usitatissimum L.).

The effect of toxic metals on seed germination was studied in 23 cultivars of flax (Linum usitatissimum L.). Toxicity of cadmium, cobalt, copper, zinc, nickel, lead, chromium, and arsenic at five different concentrations (0.01-1 mM) was tested by standard ecotoxicity test. Root length was measured after 72 h of incubation. Elongation inhibition, EC50 value, slope, and NOEC values were calculated. Results were evaluated by principal component analysis, a multidimensional statistical method. The results showed that heavy-metal toxicity decreased in the following order: As3+>or=As5+>Cu2+>Cd2+>Co2+>Cr6+>Ni2+>Pb2+>Cr3+>Zn2+.