Despite the important role of reactive oxygen species (ROS) in plant stress response, little is known about their effects on RNA. Present study focuses on RNA oxidation in soybean seedlings exposed for a short time to cadmium (1–3 h, 10 mg/L Cd). Treatment with the metal had no effect on the level of common oxidative stress markers: superoxide anion accumulation, lipid peroxidation, protein carbonylation and reduced/oxidized glutathione ratio. On the other hand, 2 h-long exposure to Cd induced the formation of the most frequent oxidative modification of RNA, 8-hydroxyguanosine (8-OHG). The immunohistochemical detection of 8-OHG in the root cross sections demonstrated that this modification is accumulated in the inner cortex tissue, endodermis, protoxylem and phloem, and co-localizes with ROS. The sequencing of 8-OHG enriched poly(A)RNA revealed that transcripts differ in their susceptibility to the oxidation. The highly oxidized transcripts (HOTs) are associated with signalling, stress response and proteins folding. In turn, the lowly oxidized transcripts (LOTs) are involved in mitochondria functioning, response to stimuli and gibberellic acid (GA) metabolism.

Plant exposure to metals is associated with the accumulation of reactive oxygen species, which mediate the oxidation of various molecules including lipids, proteins, and nucleic acids. The aim of the present study is the evaluation of the impact of short-term Cu and Pb treatment on oxidative events in the roots of soybean seedlings, with special emphasis on RNA oxidation. The results show that an increase in total RNA oxidative modification, 8-hydroxyguanosine (8-OHG), constitutes a very early response to both applied metals, observed already within the first hour of treatment. Exposure to Cu and Pb resulted also in the increase in superoxide anion and hydrogen peroxide levels and intensified lipid peroxidation. However, these responses were most prominent after longer treatment times. On the other hand, no changes were observed in the level of protein carbonylation. It can be concluded that 8-OHG enrichment in total RNA constitutes one of the earliest reactions to metals, which precedes the symptoms of oxidative stress.

Oxidation of RNA is associated with the development of numerous disorders including Alzheimer's and Parkinson's diseases, amyotrophic lateral sclerosis (ALS), cancer, and diabetes. Additionally, a correlation has been found between increase in RNA oxidation and the process of aging. In plants, elevated level of oxidatively modified transcripts has been detected during alleviation of seeds dormancy and stress response. Increasing interest on the topic of RNA oxidative modifications requires elaboration of new laboratory techniques. So far, the most common method used for the assessment of RNA oxidation is quantification of 8-hydroxyguanine (8-OHG). However, reactive oxygen species (ROS) induce also numerous other changes in nucleic acids, including formation of abasic sites (AP-sites). Recently, the level of AP-sites in RNA has been measured with the use Aldehyde Reactive Probe (ARP). In the present chapter, we describe application of this technique for the evaluation of the level of AP-sites in plant transcripts.

Iron deficiency in human diet is a serious problem leading, among others, to the development of iron-dependent anaemia, which affects 1.8 billion people worldwide. Biofortification of crop plants, including edible sprouts, is a promising strategy for combating dietary deficiencies. The aim of the present study is the evaluation of the effect of seeds imbibition in FeCl2 solution with Fe at the concentrations of 100 and 500 mgL−1 on the seeds germination, growth of the seedlings, their antioxidant activity and the level of total phenolics and flavonoids. The results show that pre-treatment of seeds with Fe leads to increased accumulation of this metal in the germinated seedlings. In addition, it affects germs antioxidant activity and the level of flavonoids. On the other hand, seeds imbibition in FeCl2 had no effect on the germination rate, growth of seedlings or total phenolic content (TPC). The results indicate that seeds imbibition in Fe solution is a promising strategy for obtaining Fe-enriched soybean sprouts.

Numerous reports find that Cd induces formation of reactive oxygen species (ROS) in plants. However, a general ROS pool is usually studied, without distinction of their production site. In the present study, we applied a mitochondria-specific antioxidant, MitoTEMPO, to elucidate the role of mitochondria-derived ROS in the response of soybean seedlings to short-term (48 h) Cd stress. The obtained results showed that Cd caused a reduction in root length and fresh weight and increase in the level of superoxide anion, hydrogen peroxide, markers of lipid peroxidation (thiobarbituric reactive substances, TBARS) and markers of RNA oxidation (8-hydroxyguanosine, 8-OHG) in seedling roots. Application of MitoTEMPO affected Cd uptake in a dose-dependent manner and diminished the Cd-dependent induction of superoxide anion and lipid peroxidation.

The survival of cells depends on their ability to replicate correctly genetic material. Cells exposed to replication stress can experience a number of problems that may lead to deregulated proliferation, the development of cancer, and/or programmed cell death. In this article, we have induced prolonged replication arrest via hydroxyurea (HU) treatment and also premature chromosome condensation (PCC) by co-treatment with HU and caffeine (CF) in the root meristem cells of Vicia faba. We have analyzed the changes in the activities of retinoblastoma-like protein (RbS807/811ph). Results obtained from the immunocytochemical detection of RbS807/811ph allowed us to distinguish five unique activity profiles of pRb. We have also performed detailed 3D modeling using Blender 2.9.1., based on the original data and some final conclusions. 3D models helped us to visualize better the events occurring within the nuclei and acted as a high-resolution aid for presenting the results. We have found that, despite the decrease in pRb activity, its activity profiles were mostly intact and clearly recognizable, with some local alterations that may correspond to the increased demand in transcriptional activity. Our findings suggest that Vicia faba's ability to withstand harsh environments may come from its well-developed and highly effective response to replication stress.

Contamination of the environment with metals, their adverse impact on plant performance and transmission to the human food chain through crops and vegetables are important concerns worldwide. Although the literature on metal contamination, toxicity and plant response to this stress factor is quite abundant, there are very limited reports on the phenomenon of plant recovery after metal stress. The present article reviews available literature on the recovery process examined in various plant species, in response to several metals (Al, Cd, Cu, Ni, Pb, Zn), applied at different concentrations and treatment duration. The reviewed studies have been carried out in laboratory conditions. However, it should be highlighted that although metal stress is not as transient as most of other stress factors (e.g., drought, heat, chilling), metal concentration in the soil may still decrease due to, e.g., leaching to lower soil layers or uptake by organisms. Thus, in natural conditions, plants may be subjected to post-metal-stress conditions. The review also discusses the mechanism behind efficient recovery and the impact of post metal stress on future plant performance-possible acquisition of stress memory, adaptation to unfavorable conditions and cross-tolerance towards other stress factors.

Heavy metal pollution causes many soils to become a toxic environment not only for plants, but also microorganisms; however, little is known how heavy metal contaminated environment affects metabolism of phytopathogens and their capability of infecting host plants. In this study the oomycete Phytophthora infestans (Mont.) de Bary, the most harmful pathogen of potato, growing under moderate cadmium stress (Cd, 5 mg/L) showed nitro-oxidative imbalance associated with an enhanced antioxidant response. Cadmium notably elevated the level of nitric oxide, superoxide and peroxynitrite that stimulated nitrative modifications within the RNA and DNA pools in the phytopathogen structures. In contrast, the protein pool undergoing nitration was diminished confirming that protein tyrosine nitration is a flexible element of the oomycete adaptive strategy to heavy metal stress. Finally, to verify whether Cd is able to modify P. infestans pathogenicity, a disease index and molecular assessment of disease progress were analysed indicating that Cd stress enhanced aggressiveness of vr P. infestans towards various potato cultivars. Taken together, Cd not only affected hyphal growth rate and caused biochemical changes in P. infestans structures, but accelerated the pathogenicity as well. The nitro-oxidative homeostasis imbalance underlies the phytopathogen adaptive strategy and survival in the heavy metal contaminated environment.

Seed germination is the earliest process in plant development and is crucial for further plant growth and fitness. The process is regulated by various internal and external factors, including soil pollutants such as nonessential metals. In the present study, we examined in detail the impact of short-term imbibition in Cd solutions at several concentrations (5, 10, and 25 mg/L) on germination rate and physiological state of soybean seeds. The results showed that although Cd was readily absorbed by the seeds, the metal had no effect on seeds cell viability, oxidative stress intensity, or germination percentage. In contrast, imbibition in Cd solution led to slight reduction in antioxidant capacity of seeds. Seedlings grown from seeds pretreated with metal showed no differences in growth in relation to the control Taken together, the results indicate that soybean seeds are relatively tolerant even to high Cd concentration (up to 25 mg/L).

BACKGROUND:Cadmium is a non-essential heavy metal, which is toxic even in relatively low concentrations. Although the mechanisms of Cd toxicity are well documented, there is limited information concerning the recovery of plants after exposure to this metal. METHODS:The present study describes the recovery of soybean plants treated for 48 h with Cd at two concentrations: 10 and 25 mg/L. In the frame of the study the growth, cell viability, level of membrane damage makers, mineral content, photosynthesis parameters, and global methylation level have been assessed directly after Cd treatment and/or after 7 days of growth in optimal conditions. RESULTS:The results show that exposure to Cd leads to the development of toxicity symptoms such as growth inhibition, increased cell mortality, and membrane damage. After a recovery period of 7 days, the exposed plants showed no differences in relation to the control in all analyzed parameters, with an exception of a slight reduction in root length and changed content of potassium, magnesium, and manganese. CONCLUSIONS:The results indicate that soybean plants are able to efficiently recover even after relatively severe Cd stress. On the other hand, previous exposure to Cd stress modulated their mineral uptake.

Abstract Background Precise regulation of gene expression is indispensable for the proper functioning of organisms in both optimal and challenging conditions. The most commonly known regulative mechanisms include the modulation of transcription, translation and adjustment of the transcript, and protein half-life. New players have recently emerged in the arena of gene expression regulators – chemical modifications of mRNAs. Main text The latest studies show that modified ribonucleotides affect transcript splicing, localization, secondary structures, interaction with other molecules and translation efficiency. Thus far, attention has been focused mostly on the most widespread mRNA modification – adenosine methylation at the N6 position (m6A). However, initial reports on the formation and possible functions of other modified ribonucleotides, such as cytosine methylated at the 5′ position (m5C), 8-hydroxyguanosine (8-OHG) and 8-nitroguanosine (8-NO2G), have started to appear in the literature. Additionally, some reports indicate that pseudouridine (Ψ) is present in mRNAs and might perform important regulatory functions in eukaryotic cells. The present review summarizes current knowledge regarding the above-mentioned modified ribonucleotides (m6A, m5C, 8-OHG, 8-NO2G) in transcripts across various plant species, including Arabidopsis, rice, sunflower, wheat, soybean and potato. Conclusions Chemical modifications of ribonucleotides affect mRNA stability and translation efficiency. They thus constitute a newly discovered layer of gene expression regulation and have a profound effect on the development and functioning of various organisms, including plants.

Lettuce (Lactuca sativa L.) is an important dietary component and its biofortification in selenium (Se) may have health-promoting effects for humans. Selenium fertilization may change plant resistance to abiotic and biotic stresses. The aim of this study was to determine the effects of Se (IV) has on lettuce resistance to biotic stress induced by Botrytis cinerea (gray mold). The results have been discussed in relation to: (1) the potential use of Se for supplementation of crops to counteract Se deficiency in diet and (2) to the effect of Se-enrichment on the susceptibility of plants to pathogens.

Increase in the level of reactive oxygen species (ROS) is a common response to stress factors, including exposure to metals. ROS over-production is associated with oxidation of lipids, proteins, and nucleic acids. It is suggested that the products of oxidation are not solely the markers of oxidative stress but also signaling elements. For instance, it has been shown in animal models that mRNA oxidation is a selective process engaged in post-transcriptional regulation of genes expression and that it is associated with the development of symptoms of several neurodegenerative disorders. In the present study, we examined the impact of short-term cadmium (Cd) stress on the level of two RNA oxidation markers: 8-hydroxyguanosine (8-OHG) and apurinic/apyrimidinic sites (AP-sites, abasic sites). In the case of 8-OHG, a significant increase was observed after 3 h of exposure to moderate Cd concentration (10 mg/l). In turn, high level of AP-sites, accompanied by strong ROS accumulation and lipid peroxidation, was noted only after 24 h of treatment with higher Cd concentration (25 mg/l). This is the first report showing induction of RNA oxidations in plants response to stress factors. The possible signaling and gene regulatory role of oxidatively modified transcripts is discussed.

In this paper we analyzed β-aminobutyric acid (BABA)-primed epigenetic adjustment of potato cv. “Sarpo Mira” to Phytophthora infestans. The first stress-free generation of the potato genotype obtained from BABA-primed parent plants via tubers and seeds showed pronounced resistance to the pathogen, which was tuned with the transcriptional memory of SA-responsive genes. During the early priming phase before the triggering stress, we found robust bistable deposition of histone marks (H3K4me2 and H3K27me3) on the NPR1 (Non-expressor of PR genes) and the SNI1 gene (Suppressor of NPR1, Inducible), in which transcription antagonized silencing. Switchable chromatin states of these adverse systemic acquired resistance (SAR) regulators probably reprogrammed responsiveness of the PR1 and PR2 genes and contributed to stress imprinting. The elevated levels of heritable H3K4me2 tag in the absence of transcription on SA-dependent genes in BABA-primed (F0) and its vegetative and generative progeny (F1) before pathogen challenge provided evidence for the epigenetic mark for intergenerational memory in potato. Moreover, our study revealed that histone acetylation was not critical for maintaining BABA-primed defense information until the plants were triggered with the virulent pathogen when rapid and boosted PRs gene expression probably required histone acetyltransferase (HAT) activity both in F0 and F1 progeny.

Iron (Fe) and magnesium (Mg) deficiency in human diets is a widespread problem observed in various regions of the world. Insufficient Fe uptake results in the development of iron dependent anaemia and depressed physical and intellectual performance. In turn Mg deficiency is associated with alterations in neuromuscular and cardiovascular systems. An emerging alternative to traditional supplementation of these elements in the form of pills, liquids or effervescent tablets, is introduction of fortified food products. In present study we show that preincubation of soybean seeds in Fe and Mg solutions leads to elevated content of these elements in the seedlings. Importantly the pretreatment did not affect germination rate, seedlings growth or, with an exception of Fe supplementation at highest concentration, antioxidant capacity. The obtained results indicate that preincubation of seeds in Fe and Mg solutions may be a promising method of obtaining enriched soybean sprouts.

Cadmium-induced oxidative burst is partially mediated by NADPH oxidase. The aim of the present research was to evaluate the role of NADPH oxidase in soybeans’ response to short-term cadmium stress. The application of an NADPH oxidase inhibitor, diphenyleneiodonium chloride (DPI), affected expression of two Cd-inducible genes, encoding DOF1 and MYBZ2 transcription factors. This effect was observed after 3 h of treatment. Interestingly, Cd-dependent increases in NADPH oxidase activity occurred only after a period of time ranging from 6 and 24 h of stress. Stimulation of the enzyme correlated in time with a significant accumulation of reactive oxygen species (ROS). Further analysis revealed that pharmacological inhibition of NADPH oxidase activity during 24 h of Cd stress does not affect Cd uptake, seedling growth, or the level of lipid peroxidation. The role of NADPH oxidase in the response of soybean seedlings to short-term Cd exposure is discussed.

Reactive oxygen species (ROS) are engaged in several processes essential for normal cell functioning, such as differentiation, anti-microbial defense, stimulus sensing and signaling. Interestingly, recent studies imply that cellular signal transduction and gene regulation are mediated not only directly by ROS but also by the molecules derived from ROS-mediated oxidation. Lipid peroxidation leads to non-enzymatic formation of oxylipins. These molecules were shown to modulate expression of signaling associated genes including genes encoding phosphatases, kinases and transcription factors. Oxidized peptides derived from protein oxidation might be engaged in organelle-specific ROS signaling. In turn, oxidation of particular mRNAs leads to decrease in the level of encoded proteins and thus, contributes to the post-transcriptional regulation of gene expression. Present mini review summarizes latest findings concerning involvement of products of lipid, protein and RNA oxidation in signal transduction and gene regulation.