Silicon's Influence on Polyphenol and Flavonoid Profiles in Pea (Pisum sativum L.) under Cadmium Exposure in Hydroponics: A Study of Metabolomics, Extraction Efficacy, and Antimicrobial Properties of Extracts.

The current study aimed to investigate the impact of silicon (Si) supplementation in the form of Na2SiO3 on the metabolome of peas under normal conditions and following exposure to cadmium (Cd) stress. Si is known for its ability to enhance stress tolerance in various plant species, including the mitigation of heavy metal toxicity. Cd, a significant contaminant, poses risks to both human health and the environment. The study focused on analyzing the levels of bioactive compounds in different plant parts, including the shoot, root, and pod, to understand the influence of Si supplementation on their biosynthesis. Metabolomic analysis of pea samples was conducted using a targeted HPLC/MS approach, enabling the identification of 15 metabolites comprising 9 flavonoids and 6 phenolic acids. Among the detected compounds, flavonoids, such as flavon and quercetin, along with phenolic acids, including chlorogenic acid and salicylic acid, were found in significant quantities. The study compared Si supplementation at concentrations of 1 and 2 mM, as well as Cd stress conditions, to evaluate their effects on the metabolomic profile. Additionally, the study explored the extraction efficiency of three different methods: accelerated solvent extraction (ASE), supercritical fluid extraction (SFE), and maceration (MAC). The results revealed that SFE was the most efficient method for extracting polyphenolic compounds from the pea samples. Moreover, the study investigated the stability of polyphenolic compounds under different pH conditions, ranging from 4.0 to 6.0, providing insights into the influence of the pH on the extraction and stability of bioactive compounds.

Non-photochemical quenching upregulation improves water use efficiency and reduces whole plant level water consumption under drought.

For crop production, the water supply limitations will likely become a bigger restriction underlining a need for crops that use less water per mass of production. Therefore, water use efficiency becomes a constraint in obtaining resilient and productive crops. We hypothesized that under drought conditions via modulation of chloroplast signal for stomatal opening by upregulation of non-photochemical quenching (NPQ) it is possible to improve water use efficiency. Nicotiana tabacum plants with strong overexpression of photosystem II subunit S (PsbS), a key protein to NPQ, were grown under varied levels of drought. The PsbS-overexpressing lines lost 11% less water per CO2 fixed under drought which did not have a significant effect on plant size. Depending on growth conditions PsbS-overexpressing lines on the whole plant level consumed from 4% to 30% less water than the corresponding wildtype. The leaf water and chlorophyll contents showed a positive relation with the level of NPQ. Our study provides proof of concept and as such is an important step towards engineering crops with improved water use efficiency.

Metal tolerance and Cd phytoremoval ability in Pisum sativum grown in spiked nutrient solution.

In the presented study, the effects of cadmium (Cd) stress and silicon (Si) supplementation on the pea plant (Pisum sativum L.) were investigated. The tendency to accumulate cadmium in the relevant morphological parts of the plant (roots and shoots respectively)-bioaccumulation, the transfer of this element in the plant (translocation) and the physiological parameters of the plant through indicators of oxidative stress were determined. Model studies were carried out at pH values 6.0 and 5.0 plant growth conditions in the hydroponic cultivation. It was shown that Cd accumulates mostly in plant roots at both pH levels. However, the Cd content is higher in the plants grown at lower pH. The Cd translocation factor was below 1.0, which indicates that the pea is an excluder plant. The contamination of the plant growth environment with Cd causes the increased antioxidant stress by the growing parameters of the total phenolic content (TPC), polyphenol oxidase activity (PPO), the malondialdehyde (MDA) and lipid peroxidation (LP). The results obtained showed that the supplementation with Si reduces these parameters, thus lowering the oxidative stress of the plant. Moreover, supplementation with Si leads to a lower content of Cd in the roots and reduces bioaccumulation of Cd in shoots and roots of pea plants.

Is chloroplast size optimal for photosynthetic efficiency?

Improving photosynthetic efficiency has recently emerged as a promising way to increase crop production in a sustainable manner. While chloroplast size may affect photosynthetic efficiency in several ways, we aimed to explore whether chloroplast size manipulation can be a viable approach to improving photosynthetic performance. Several tobacco (Nicotiana tabacum) lines with contrasting chloroplast sizes were generated via manipulation of chloroplast division genes to assess photosynthetic performance under steady-state and fluctuating light. A selection of lines was included in a field trial to explore productivity. Lines with enlarged chloroplasts underperformed in most of the measured traits. Lines with smaller and more numerous chloroplasts showed a similar efficiency compared with wild-type (WT) tobacco. Chloroplast size only weakly affected light absorptance and light profiles within the leaf. Increasing chloroplast size decreased mesophyll conductance (gm ) but decreased chloroplast size did not increase gm . Increasing chloroplast size reduced chloroplast movements and enhanced non-photochemical quenching. The chloroplast smaller than WT appeared to be no better than WT for photosynthetic efficiency and productivity under field conditions. The results indicate that chloroplast size manipulations are therefore unlikely to lead to higher photosynthetic efficiency or growth.

Comprehensive Study of Si-Based Compounds in Selected Plants (Pisum sativum L., Medicago sativa L., Triticum aestivum L.).

This review describes the role of silicon (Si) in plants. Methods of silicon determination and speciation are also reported. The mechanisms of Si uptake by plants, silicon fractions in the soil, and the participation of flora and fauna in the Si cycle in terrestrial ecosystems have been overviewed. Plants of Fabaceae (especially Pisum sativum L. and Medicago sativa L.) and Poaceae (particularly Triticum aestivum L.) families with different Si accumulation capabilities were taken into consideration to describe the role of Si in the alleviation of the negative effects of biotic and abiotic stresses. The article focuses on sample preparation, which includes extraction methods and analytical techniques. The methods of isolation and the characterization of the Si-based biologically active compounds from plants have been overviewed. The antimicrobial properties and cytotoxic effects of known bioactive compounds obtained from pea, alfalfa, and wheat were also described.

Genetic control of photoprotection and photosystem II operating efficiency in plants.

Photoprotection against excess light via nonphotochemical quenching (NPQ) is indispensable for plant survival. However, slow NPQ relaxation under low light conditions can decrease yield of field-grown crops up to 40%. Using semi-high-throughput assay, we quantified the kinetics of NPQ and photosystem II operating efficiency (ΦPSII) in a replicated field trial of more than 700 maize (Zea mays) genotypes across 2 yr. Parametrized kinetics data were used to conduct genome-wide association studies. For six candidate genes involved in NPQ and ΦPSII kinetics in maize the loss of function alleles of orthologous genes in Arabidopsis (Arabidopsis thaliana) were characterized: two thioredoxin genes, and genes encoding a transporter in the chloroplast envelope, an initiator of chloroplast movement, a putative regulator of cell elongation and stomatal patterning, and a protein involved in plant energy homeostasis. Since maize and Arabidopsis are distantly related, we propose that genes involved in photoprotection and PSII function are conserved across vascular plants. The genes and naturally occurring functional alleles identified here considerably expand the toolbox to achieving a sustainable increase in crop productivity.

A method for reducing the concentrations of Fusarium graminearum trichothecenes in durum wheat grain with the use of Debaryomyces hansenii.

Fusarium head blight (FHB), caused mainly by Fusarium graminearum, is one of the most dangerous diseases of durum wheat. This hemibiotrophic pathogen transitions from the biotrophic phase, during which it penetrates host tissues and secretes trichothecenes, to the necrotrophic phase which leads to the destruction of host tissues. Yeasts applied to spikes often reduce mycotoxin concentrations, but the underlying mechanisms have not been fully elucidated. Therefore, the aim of this study was to analyze the concentrations trichothecenes in durum wheat grain and changes in the F. graminearum transcriptome under the influence the Debaryomyces hansenii antagonistic yeast strain. Debaryomyces hansenii cells adhered to and formed cell aggregates/biofilm on the surface of spikes and pathogenic hyphae. Biological control suppressed the spread of F. graminearum by 90 % and decreased the content of deoxynivalenol (DON) in spikes by 31.2 %. Yeasts significantly reduced the expression of pathogen's genes encoding the rpaI subunit of RNA polymerase I and the activator of Hsp90 ATPase, but they had no effect on mRNA transcript levels of genes encoding the enzymes involved in the biosynthesis of trichothecenes. The yeast treatment reduced the number of F. graminearum operational taxonomic units (OTUs) nearly five-fold and increased the number of D. hansenii OTUs more than six-fold in the spike mycobiome. The mechanisms that suppress infections should be explored to develop effective biological methods for reducing the concentrations mycotoxins in wheat grain.

The Size-Dependent Effects of Silver Nanoparticles on Germination, Early Seedling Development and Polar Metabolite Profile of Wheat (Triticum aestivum L.).

The phytotoxicity of silver nanoparticles (Ag NPs) to plant seeds germination and seedlings development depends on nanoparticles properties and concentration, as well as plant species and stress tolerance degrees. In the present study, the effect of citrate-stabilized spherical Ag NPs (20 mg/L) in sizes of 10, 20, 40, 60, and 100 nm, on wheat grain germination, early seedlings development, and polar metabolite profile in 3-day-old seedlings were analyzed. Ag NPs, regardless of their sizes, did not affect the germination of wheat grains. However, the smaller nanoparticles (10 and 20 nm in size) decreased the growth of seedling roots. Although the concentrations of total polar metabolites in roots, coleoptile, and endosperm of seedlings were not affected by Ag NPs, significant re-arrangements of carbohydrates profiles in seedlings were noted. In roots and coleoptile of 3-day-old seedlings, the concentration of sucrose increased, which was accompanied by a decrease in glucose and fructose. The concentrations of most other polar metabolites (amino acids, organic acids, and phosphate) were not affected by Ag NPs. Thus, an unknown signal is released by small-sized Ag NPs that triggers affection of sugars metabolism and/or distribution.

Guard-cell-targeted overexpression of Arabidopsis Hexokinase 1 can improve water use efficiency in field-grown tobacco plants.

Water deficit currently acts as one of the largest limiting factors for agricultural productivity worldwide. Additionally, limitation by water scarcity is projected to continue in the future with the further onset of effects of global climate change. As a result, it is critical to develop or breed for crops that have increased water use efficiency and that are more capable of coping with water scarce conditions. However, increased intrinsic water use efficiency (iWUE) typically brings a trade-off with CO2 assimilation as all gas exchange is mediated by stomata, through which CO2 enters the leaf while water vapor exits. Previously, promising results were shown using guard-cell-targeted overexpression of hexokinase to increase iWUE without incurring a penalty in photosynthetic rates or biomass production. Here, two homozygous transgenic tobacco (Nicotiana tabacum) lines expressing Arabidopsis Hexokinase 1 (AtHXK1) constitutively (35SHXK2 and 35SHXK5) and a line that had guard-cell-targeted overexpression of AtHXK1 (GCHXK2) were evaluated relative to wild type for traits related to photosynthesis and yield. In this study, iWUE was significantly higher in GCHXK2 compared with wild type without negatively impacting CO2 assimilation, although results were dependent upon leaf age and proximity of precipitation event to gas exchange measurement.

The Effect of Bio-Synthesized Silver Nanoparticles on Germination, Early Seedling Development, and Metabolome of Wheat (Triticum aestivum L.).

Changes in the metabolome of germinating seeds and seedlings caused by metal nanoparticles are poorly understood. In the present study, the effects of bio-synthesized silver nanoparticles ((Bio)Ag NPs) on grains germination, early seedlings development, and metabolic profiles of roots, coleoptile, and endosperm of wheat were analyzed. Grains germinated well in (Bio)Ag NPs suspensions at the concentration in the range 10-40 mg/L. However, the growth of coleoptile was inhibited by 25%, regardless of (Bio)Ag NPs concentration tested, whereas the growth of roots gradually slowed down along with the increasing concentration of (Bio)Ag NPs. The deleterious effect of Ag NPs on roots was manifested by their shortening, thickening, browning of roots tips, epidermal cell death, progression from apical meristem up to root hairs zone, and the inhibition of root hair development. (Bio)Ag NPs stimulated ROS production in roots and affected the metabolic profiles of all tissues. Roots accumulated sucrose, maltose, 1-kestose, phosphoric acid, and some amino acids (i.e., proline, aspartate/asparagine, hydroxyproline, and branched-chain amino acids). In coleoptile and endosperm, contrary to roots, the concentration of most metabolites decreased. Moreover, coleoptile accumulated galactose. Changes in the concentration of polar metabolites in seedlings revealed the affection of primary metabolism, disturbances in the mobilization of storage materials, and a translocation of sugars and amino acids from the endosperm to growing seedlings.

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.

Root NRT, NiR, AMT, GS, GOGAT and GDH expression levels reveal NO and ABA mediated drought tolerance in Brassica juncea L.

Little is known about the interactive effects of exogenous nitric oxide (NO) and abscisic acid (ABA) on nitrogen (N) metabolism and related changes at molecular and biochemical levels under drought stress. The present study highlights the independent and combined effect of NO and ABA (grouped as "nitrate agonists") on expression profiles of representative key genes known to be involved in N-uptake and assimilation, together with proline metabolism, N-NO metabolism enzyme's activity and nutrient content in polyethylene glycol (PEG) treated roots of Indian mustard (B. juncea cv. Varuna). Here we report that PEG mediated drought stress negatively inhibited growth performance, as manifested by reduced biomass (fresh and dry weight) production. Total N content and other nitrogenous compounds (NO3-, NO2-) were decreased; however, NH4+, NH4+/ NO3- ratio and total free amino acids content were increased. These results were positively correlated with the PEG induced changes in expression of genes and enzymes involved in N-uptake and assimilation. Also, PEG supply lowered the content of macro- and micro-nutrients but proline level and the activity of ∆1-pyrroline-5-carboxylate synthetase increased indicating increased oxidative stress. However, all these responses were reversed upon the exogenous application of nitrate agonists (PEG + NO, PEG + NO + ABA, and PEG + ABA) where NO containing nitrate agonist treatment i.e. PEG + NO was significantly more effective than PEG + ABA in alleviating drought stress. Further, increases in activities of L-arginine dependent NOS-like enzyme and S-nitrosoglutathione reductase were observed under nitrate agonist treatments. This indicates that the balanced endogenous change in NO and ABA levels together during synthesis and degradation of NO mitigated the oxidative stress in Indian mustard seedlings. Overall, our results reveal that NO independently or together with ABA may contribute to improved crop growth and productivity under drought stress.

Polypropylene structure alterations after 5 years of natural degradation in a waste landfill.

Up to 25% of plastic waste in Europe is still disposed of in landfills, despite recycling efforts. The plastic waste in the landfill plot may be exposed both to abiotic and biotic degradation processes, although it is thought that most of the plastic materials tend to be resistant to biodegradation or biodeterioration even after a long time. To verify if polypropylene (PP) can undergo the process of short-term biodegradation and how this process is manifested in a municipal waste landfill, we collected a plastic sample from an already closed landfill plot estimating its age at approximately 5 years. Fourier-Transform Infrared Spectroscopy led to sample identification as PP as well as showed additional bands which are not specific to polymer structure but rather result from microbial metabolism. Differential Scanning Calorimetry was performed to examine the influence of the environmental degradation process on the degree of crystallisation of the tested PP. Moreover, significant changes on the surface of an old PP packaging were observed with Scanning Electron Microscopy (SEM) showing clear signs of PP delamination which resulted in microplastic particles formation (smaller than 5 µm in diameter). Additionally, several round and oval shaped structures were observed with SEM leading to the suspicion of biofilm formation on the PP surface. Indeed, the microorganisms were present in a vast amount on the old PP surface and possibly formed a viable biofilm as it was confirmed with fluorescence microscopy. These data show that plastic waste can be inhabited by microorganisms from the ambient environment which may probably lead to its faster degradation. However, this process should be investigated in more detail in order to shed light upon the possible risk factors of plastic biodegradation in waste landfills to the environment and human health. Even after five years, polypropylene can undergo deterioration/biodegradation in a waste landfill with viable microbial cells on its surface, possibly involved in its degradation.

Genome biology of the paleotetraploid perennial biomass crop Miscanthus.

Miscanthus is a perennial wild grass that is of global importance for paper production, roofing, horticultural plantings, and an emerging highly productive temperate biomass crop. We report a chromosome-scale assembly of the paleotetraploid M. sinensis genome, providing a resource for Miscanthus that links its chromosomes to the related diploid Sorghum and complex polyploid sugarcanes. The asymmetric distribution of transposons across the two homoeologous subgenomes proves Miscanthus paleo-allotetraploidy and identifies several balanced reciprocal homoeologous exchanges. Analysis of M. sinensis and M. sacchariflorus populations demonstrates extensive interspecific admixture and hybridization, and documents the origin of the highly productive triploid bioenergy crop M. × giganteus. Transcriptional profiling of leaves, stem, and rhizomes over growing seasons provides insight into rhizome development and nutrient recycling, processes critical for sustainable biomass accumulation in a perennial temperate grass. The Miscanthus genome expands the power of comparative genomics to understand traits of importance to Andropogoneae grasses.

Training Population Optimization for Genomic Selection in Miscanthus.

Miscanthus is a perennial grass with potential for lignocellulosic ethanol production. To ensure its utility for this purpose, breeding efforts should focus on increasing genetic diversity of the nothospecies Miscanthus × giganteus (M×g) beyond the single clone used in many programs. Germplasm from the corresponding parental species M. sinensis (Msi) and M. sacchariflorus (Msa) could theoretically be used as training sets for genomic prediction of M×g clones with optimal genomic estimated breeding values for biofuel traits. To this end, we first showed that subpopulation structure makes a substantial contribution to the genomic selection (GS) prediction accuracies within a 538-member diversity panel of predominately Msi individuals and a 598-member diversity panels of Msa individuals. We then assessed the ability of these two diversity panels to train GS models that predict breeding values in an interspecific diploid 216-member M×g F2 panel. Low and negative prediction accuracies were observed when various subsets of the two diversity panels were used to train these GS models. To overcome the drawback of having only one interspecific M×g F2 panel available, we also evaluated prediction accuracies for traits simulated in 50 simulated interspecific M×g F2 panels derived from different sets of Msi and diploid Msa parents. The results revealed that genetic architectures with common causal mutations across Msi and Msa yielded the highest prediction accuracies. Ultimately, these results suggest that the ideal training set should contain the same causal mutations segregating within interspecific M×g populations, and thus efforts should be undertaken to ensure that individuals in the training and validation sets are as closely related as possible.

Photosynthetic efficiency and mesophyll conductance are unaffected in Arabidopsis thaliana aquaporin knock-out lines.

Improving photosynthetic efficiency is widely regarded as a major route to achieving much-needed yield gains in crop plants. In plants with C3 photosynthesis, increasing the diffusion conductance for CO2 transfer from substomatal cavity to chloroplast stroma (gm) could help to improve the efficiencies of CO2 assimilation and photosynthetic water use in parallel. The diffusion pathway from substomatal cavity to chloroplast traverses cell wall, plasma membrane, cytosol, chloroplast envelope membranes, and chloroplast stroma. Specific membrane intrinsic proteins of the aquaporin family can facilitate CO2 diffusion across membranes. Some of these aquaporins, such as PIP1;2 in Arabidopsis thaliana, have been suggested to exert control over gm and the magnitude of the CO2 assimilation flux, but the evidence for a direct physiological role of aquaporins in determining gm is limited. Here, we estimated gm with four different methods under a range of light intensities and CO2 concentrations in two previously characterized pip1;2 knock-out lines as well as pip1;3 and pip2;6 knock-out lines, which have not been previously evaluated for a role in gm. This study presents the most in-depth analysis of gm in Arabidopsis aquaporin knock-out mutants to date. Surprisingly, all methods failed to show any significant differences between the pip1;2, pip1;3, or pip2;6 mutants and the Col-0 control.

The Effect of Cadmium on the Activity of Stress-Related Enzymes and the Ultrastructure of Pea Roots.

The analysis of the effects of cadmium (Cd) on plant cells is crucial to understand defense mechanisms and adaptation strategies of plants against Cd toxicity. In this study, we examined stress-related enzyme activities after one and seven days of Cd application and the ultrastructure of roots of Pisum sativum L. after seven days of Cd treatment (10, 50, 100, and 200 µM CdSO4). Our results showed that phenylalanine ammonia-lyase (PAL) activity and the amount of Cd accumulated in the roots were significantly positively correlated with the Cd concentration used in our experiment. However, Cd caused a decrease of all studied antioxidative enzyme activities (i.e., catalase (CAT), ascorbate peroxidase (APX), guaiacol peroxidase (GPX)). The analysis of the ultrastructure (TEM) showed various responses to Cd, depending on Cd concentrations. In general, lower Cd concentrations (50 and 100 µM CdSO4) mostly resulted in increased amounts of oil bodies, plastolysomes and the accumulation of starch granules in plastids. Meanwhile, roots treated with a higher concentration of Cd (200 µM CdSO4) additionally triggered protective responses such as an increased deposition of suberin lamellae in the endodermal cell walls. This indicates that Cd induces a complex defense response in root tissues.

The Associations between Leaf Morphology, Phenylalanine Ammonia Lyase Activity, Reactive Oxygen Species, and Fusarium Resistance in Selected Species of Wheat with Different Ploidy Levels.

In wheat, resistance to Fusarium is conditioned by anatomical, morphological, and physiological traits. The aim of this study was to evaluate selected elements of constitutive barriers in common wheat, spelt, Polish wheat, emmer, and einkorn. The activity of the phenylalanine ammonia-lyase (PAL) enzyme and rate of reactive oxygen species (ROS) production were evaluated in the tissues of common wheat and spelt inoculated with Fusarium culmorum. Most of the relict wheat species were more abundant in morphological barriers than common wheat. F. culmorum penetrated constitutive barriers, which increased PAL activity and intensified ROS production 24 h after inoculation in wheat tissues. The lowest increase in PAL activity after inoculation was observed in cv. Sumai3, which resistance is based on limiting the spread of F. culmorum within the spike. Spelt line Tas 581 glumes were characterized by the highest concentration of ROS 24 h after inoculation. The ROS content remained high for five days. The results of this study indicate that high trichome density plays a key role in resistance to pathogens. In the resistant spelt line with effective constitutive barriers, PAL activity and ROS content were higher than those observed in susceptible wheats after inoculation with F. culmorum.

Predicting light-induced stomatal movements based on the redox state of plastoquinone: theory and validation.

Prediction of stomatal conductance is a key element to relate and scale up leaf-level gas exchange processes to canopy, ecosystem and land surface models. The empirical models that are typically employed for this purpose are simple and elegant formulations which relate stomatal conductance on a leaf area basis to the net rate of CO2 assimilation, humidity and CO2 concentration. Although light intensity is not directly modelled as a stomatal opening cue, it is well-known that stomata respond strongly to light. One response mode depends specifically on the blue-light part of the light spectrum, whereas the quantitative or 'red' light response is less spectrally defined and relies more on the quantity of incident light. Here, we present a modification of an empirical stomatal conductance model which explicitly accounts for the stomatal red-light response, based on a mesophyll-derived signal putatively initiated by the chloroplastic plastoquinone redox state. The modified model showed similar prediction accuracy compared to models using a relationship between stomatal conductance and net assimilation rate. However, fitted parameter values with the modified model varied much less across different measurement conditions, lessening the need for frequent re-parameterization to different conditions required of the current model. We also present a simple and easy to parameterize extension to the widely used Farquhar-Von Caemmerer-Berry photosynthesis model to facilitate coupling with the modified stomatal conductance model, which should enable use of the new stomatal conductance model to simulate ecosystem water vapour exchange in terrestrial biosphere models.

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.