Changes in Metabolism and Content of Chlorophyll in Common Duckweed (Lemna minor L.) Caused by Environmental Contamination with Fluorides.

The impact of fluorine on plants remains poorly understood. We examined duckweed growth in extracts of soil contaminated with fluorine leached from chicken manure. Additionally, fluorine levels were analyzed in fresh manure, outdoor-stored manure, and soil samples at varying distances from the manure pile. Fresh manure contained 37-48 mg F- × kg-1, while soil extracts contained 2.1 to 4.9 mg F- × kg-1. We evaluated the physiological effects of fluorine on duckweed cultured on soil extracts or in 50% Murashige-Skoog (MS) medium supplemented with fluorine concentrations matching those in soil samples (2.1 to 4.9 mg F- × L-1), as well as at 0, 4, and 210 mg × L-1. Duckweed exposed to fluorine displayed similar toxicity symptoms whether in soil extracts or supplemented medium. Fluoride at concentrations of 2.1 to 4.9 mg F- × L-1 reduced the intact chlorophyll content, binding the porphyrin ring at position 32 without affecting Mg2+. This reaction resulted in chlorophyll a absorption peak shifted towards shorter wavelengths and formation of a new band of the F--chlorophyll a complex at λ = 421 nm. Moreover, plants exposed to low concentrations of fluorine exhibited increased activities of aminolevulinic acid dehydratase and chlorophyllase, whereas the activities of both enzymes sharply declined when the fluoride concentration exceeded 4.9 mg × L-1. Consequently, fluorine damages chlorophyll a, disrupts the activity of chlorophyll-metabolizing enzymes, and diminishes the plant growth rate, even when the effects of these disruptions are too subtle to be discerned by the naked human eye.

Studies on the Efficiency of Iron Release from Fe(III)-EDTA and Fe(III)-Cit and the Suitability of These Compounds for Tetracycline Degradation.

Iron ions can be used to degrade tetracycline dispersed in nature. Studies of absorption and fluorescence spectra and quantum chemistry calculations showed that iron is more readily released from Fe(III)-citrate than from Fe(III)-EDTA, so Fe(III)-citrate (Fe(III)-Cit) is more suitable for tetracycline (TC) degradation. At 30 °C, a severe degradation of TC by Fe(III)-Cit occurred as early as after 3 days of incubation in the light, and after 5 days in the dark. In contrast, the degradation of TC by Fe(III)-EDTA proceeded very slowly in the dark. By the fifth day of incubation of TC with Fe(III)-Cit in darkness, the concentrations of the former compound dropped by 55% and 75%, at 20 °C and 30 °C, respectively. The decrease in tetracycline concentrations caused by Fe(III)-EDTA in darkness at the same temperatures was only 2% and 6%, respectively. Light increased the degradation rates of TC by Fe(III)-EDTA to 20% and 56% at 20 °C and 30 °C, respectively. The key role of the light in the degradation of tetracycline by Fe(III)-EDTA was thus demonstrated. The TC degradation reaction showed a second-order kinetics. The rate constants of Fe(III)-Cit-induced TC degradation at 20 °C and 30 °C in darkness were k = 4238 M-1day-1 and k = 11,330 M-1day-1, respectively, while for Fe(III)-EDTA were 55 M-1day-1 and 226 M-1day-1. In light, these constants were k = 15,440 M-1day-1 and k = 40,270 M-1day-1 for Fe(III)-Cit and k = 1012 M-1day-1 and 2050 M-1day-1 at 20 °C and 30 °C; respectively. A possible reason for the higher TC degradation rate caused by Fe(III)-Cit can be the result of its lower thermodynamical stability compared with Fe(III)-EDTA, which we confirmed with our quantum chemistry calculations. Two quantum chemistry calculations showed that the iron complex with EDTA is more stable (the free energy of the ensemble is 15.8 kcal/mol lower) than the iron complex with Cit; hence, Fe release from Fe(III)-EDTA is less effective.

Physiological and Biochemical Parameters of Common Duckweed Lemna minor after the Exposure to Tetracycline and the Recovery from This Stress.

In this study, the ability of Lemna minor L. to recover to normal growth, after being degraded in a tetracycline-containing medium, was extensively investigated. The plants were exposed to tetracycline (TC) at concentrations of 1, 2.5, and 10 mM. Subsequently, their physiological status was analysed against the following criteria: rate of plant growth; free radical accumulation; antioxidant enzyme activity; chlorophyll content; HSP70 protein content; cell membrane permeability, and mitochondrial activity. The study showed that duckweed can considerably recover from the damage caused by antibiotics, within a week of cessation of stress. Of the plant properties analysed, mitochondrial activity was the most sensitive to antibiotic-induced disturbances. After transferring the plants to a tetracycline-free medium, all plant parameters improved significantly, except for the mitochondrial activity in the plants grown on the medium containing the highest dose of tetracycline. In the plants treated with this antibiotic at the concentration of 10 mM, the proportion of dead mitochondria increased and was as high as 93% after one week from the beginning of the recovery phase, even after the transfer to the tetracycline-free medium.

Phytotoxicity of sulfamethazine soil pollutant to six legume plant species.

The effect of traces of sulfamethazine (SMZ) in soil (0.01, 0.1, 0.25, 1, 5, 15, and 20 mM) on cellular distribution of cytochrome c oxidase activity, shoot and root growth, and leachate electroconductivity was analyzed in germinating seeds of yellow lupin, pea, lentil, soybean, adzuki bean, and alfalfa. Results showed that a high activity of cytochrome c oxidase in mitochondria correlated with high seed vigor and viability. The appearance of necroses and root decay was associated with a decrease in the activity of mitochondrial cytochrome c oxidase but was accompanied by an increase in cytosolic cytochrome c oxidase activity. A short exposure period of seeds (3 and 6 d) to sulfamethazine did not influence germination. Elongation of roots and stems was more sensitive than germination rate as an indicator of soil contamination by sulfamethazine. Among all tested leguminous plants, yellow lupin was the most reliable bioindicator of SMZ contaminated soil.

A Strong Impact of Soil Tetracycline on Physiology and Biochemistry of Pea Seedlings.

Antibiotics are a new type of contaminants found in the environment. They are increasingly used in farm animal production systems and may accumulate in crops, limiting the plant growth rate and nutritive value. The aim of this study was to determine the effects of tetracycline (TC) on physiological and biochemical properties of pea seedlings. The presence of TC in the soil during 24 hours did not result in any distinct changes of the seedlings. However, after five days (120 h) of soil TC action, the seedling appearance and metabolic activities were significantly affected. Leaves lost their green coloration as a result of a 38% degradation of their chlorophyll. Total protein was isolated from shoots of pea grown for 120 h in TC-supplemented perlite (250 mg × L-1) or perlite with no TC (control plants). The 2D electrophoretic maps of proteins from non-TC shoots contained 326 spots, whereas maps of shoot proteins from TC-treated seedlings contained only 316 spots. The identity of 26 proteins was determined. The intensity of most proteins (62%) increased. This was particularly visible with diphosphate kinase, superoxide dismutase [Cu-Zn], peroxiredoxin, and glutathione S-transferase. A distinctly increased quantity of a protein involved in photosynthesis (photosystem II stability/assembly factor HCF136) was also noted. One protein was detected only in shoots of TC-treated plants (as opposed to controls); however, it could not be identified. Moreover, at the highest concentration of TC (250 mg × L-1 of perlite), a sharp increase in free-radical content was observed along with the amount of callose deposited in vascular bundles of leaves and roots and the occurrence of masses of dead cells in roots. It was found, therefore, that tetracycline which has been known for inhibiting predominantly the attachment of aminoacyl-tRNA to the ribosomal acceptor in bacteria can disturb diverse metabolic pathways in plants.

Uptake and reaction to roundup ultra 360 SL in soybean seedlings.

Due to the widespread and frequent use of Roundup Ultra 360 SL in crops production, the active substance glyphosate is often present (in the soil or in post-harvest remnants) and may be toxic to plants, including the non-target species. The aim of the current study was to determine the sensitivity of young soybean seedlings to glyphosate in concentrations ranging from 0 to 10 µM. It was demonstrated that the seedlings take small quantities of soil glyphosate up. More of the active substance was found in the shoots than in the roots. From the doses applied, the plant absorbs up to 4% of soil glyphosate, while over 96% remains in the soil. This suggests that only 4% of glyphosate taken up from the soil affects plant seedling development and water management. It modifies the contents of the biogenic amines cadaverine and putrescine as well as the activity of enzymes involved in their biosynthesis, i.e. ornithine decarboxylase and lysine decarboxylase. The free radical content of the roots increased with increasing herbicide doses and time of exposure. The main enzyme involved in the rapid removal of free radicals was superoxide peroxidase, activated by the herbicide treatment, while catalase was not significantly stimulated.

Instability of chlorophyll in yellow lupin seedlings grown in soil contaminated with ciprofloxacin and tetracycline.

With increasing soil concentrations of ciprofloxacin and tetracycline a decrease of leaf chlorophyll content was observed. Tetracycline was more detrimental than ciprofloxacin. The chlorophyll content in plants growing for ten days on a tetracycline containing soil decreased by 68%. The decrease of chlorophyll concentration was even sharper in new leaves that formed after application of the antibiotic (up to 81% drop). The comparison of absorption spectra of commercial, reagent grade chlorophyll, alone and incubated with antibiotics, has shown that ciprofloxacin and tetracycline can react directly with chlorophyll and decrease its concentration by 47.7% and 48.5%, respectively. The changes in fluorescence spectra confirmed the formation of chlorophyll degradation product. The chlorophyll decay was a second order reaction and depended on antibiotic concentration and duration of exposure. Reaction rate constants differed with antibiotics and their soil concentrations. With increasing contents of antibiotics in soil the constant of chlorophyll degradation rate in lupin plants increased from k = 870 M-1day-1 for 3 mg ciprofloxacin to k = 2490 M-1day-1 for 90 mg ciprofloxacin, and in the case of tetracycline the reaction rate constant increased from k = 1330 M-1day-1 to k = 2910 M-1day-1. The sensitivity of chlorophyll to ciprofloxacin and tetracycline was confirmed by determining EC and TU indices.