Degradation of chlorophyll was studied in leaves of spinach grown in soil containing cadmium and tetracycline, based on spectroscopic measurements and biochemical analyses of plant extracts. It was shown that plant exposure to the highest levels of tetracycline and cadmium resulted in 64% and 68%, respectively, reduction in chlorophyll content. The chlorophyll degradation rate constants were determined, and they were found to increase with increasing doses of tetracycline and cadmium. The rate constant of chlorophyll degradation by tetracycline ranged from k = 960 M⁻¹ day⁻¹ to k = 2180 M⁻¹ day⁻¹, and the rate constant of chlorophyll degradation by cadmium ranged from k = 1130 M⁻¹ day⁻¹ to k = 2580 M⁻¹ day⁻¹, depending on dose. Plant stress responses to tetracycline are much stronger than to cadmium, as is visible from the activity of guaiacol peroxidase and catalase. However, phytotoxicity of cadmium, measured by the rate of chlorophyll degradation and enzyme activities, is much higher compared to tetracycline. The spectroscopic measurements were taken 10 days after tetracycline and cadmium were added to the reagent grade chlorophyll which was used at the concentration of chlorophyll in plants. Changes in absorption and fluorescence spectra are likely to result from removal of magnesium from the chlorophyll molecule, and thus they indicate the formation of pheophytin. Cadmium, on the other hand, is probably bound into the chlorophyll molecule, substituting its magnesium.

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.

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.

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.

The widespread use of antibiotics in intensive animal husbandry, and the agricultural utilization of manure from such farms, imposes a significant burden on the environment. Consequently, the effects of antibiotics should be studied not only in animals and humans but also in all components of biocenoses and agrocenoses. In our study, we analyze the impact of four different concentrations of tetracycline present in soil (0, 5, 50, and 500 mg/kg of soil) on the growth and key photosynthesis parameters of pea seedlings: chlorophyll concentration, aminolevulinic acid concentration, aminolevulinic acid dehydrogenase activity, and ribulose bisphosphate carboxylase-oxygenase (RuBisCO) activity. At the lowest tetracycline concentration, chlorophyll content decreased by 13% compared to the control (0 tetracycline), while at the highest antibiotic concentration, it decreased by as much as 27%. Similarly, the decrease in aminolevulinic acid (a chlorophyll precursor) concentration was significant, amounting to 34%. However, the activity of the dehydrogenase enzyme, which consumes this precursor, decreased even more drastically by 51%, indicating significant disturbances in the light phase of photosynthesis. However, the activity of RuBisCO in pea plants subjected to tetracycline was even more severely affected, dropping by 58%, 69%, and 70% in soils with increasing concentrations of tetracycline. The reduction in enzyme activity could only partially be explained by a less pronounced decrease in the quantity of RuBisCO (large subunit) protein, which amounted to 6.5%, 11%, and 35% for tetracycline concentrations of 5, 50, and 500 mg/kg of soil, respectively.