Effects of drought stress memory on the accumulation of stress-protective compounds in naturally grown pine and spruce.

Forest trees are subjected to multiple stressors during their long lifetime and therefore require effective and finely regulated stress-protective systems. Stressors can induce protective systems either directly or with the involvement of stress memory mechanisms. Stress memory has only begun to be uncovered in model plants and is unexplored in coniferous species. Therefore, we studied the possible role of stress memory in the regulation of the accumulation of stress-protective compounds (heat shock proteins, dehydrins, proline) in the needles of naturally grown Scots pine and Norway spruce trees subjected to the subsequent action of long-term (multiyear) and short-term (seasonal) water shortages. Although the water deficit was relatively mild, it significantly influenced the pattern of expression of stress memory-related heat shock factor (HSF) and SWI/SNF genes, indicating the formation of stress memory in both species. In spruce, dehydrin accumulation was increased by water shortage in a manner compatible with Type II stress memory. The accumulation of HSP40 in spruce needles was positively influenced by long-term water shortage, but this increase was unlikely to be of biological importance due to the concomitant decrease in HSP70, HSP90 and HSP101 accumulation. Finally, proline accumulation was negatively influenced by short-term water deficit in spruce. In pine, no one protective compound accumulated in response to water stress. Taken together, the results indicate that the accumulation of stress-protective compounds was generally independent of stress memory effects both in pine and in spruce.

Abscisic Acid and Cytokinins Are Not Involved in the Regulation of Stomatal Conductance of Scots Pine Saplings during Post-Drought Recovery.

Delayed or incomplete recovery of gas exchange after water stress relief limits assimilation in the post-drought period and can thus negatively affect the processes of post-drought recovery. Abscisic acid (ABA) accumulation and antagonistic action between ABA and cytokinins (CKs) play an important role in regulation of stomatal conductance under water deficit. Specifically, in pine species, sustained ABA accumulation is thought to be the main cause of delayed post-drought gas exchange recovery, although the role of CKs is not yet known. Therefore, we aimed to study the effects of ABA and CKs on recovery of stomatal conductance in greenhouse-grown 3-year-old Scots pine saplings recovering from water stress. We analysed both changes in endogenous ABA and CK contents and the effects of treatment with exogenous CK on stomatal conductance. Drought stress suppressed stomatal conductance, and post-drought stomatal conductance remained suppressed for 2 weeks after plant rewatering. ABA accumulated during water stress, but ABA levels decreased rapidly after rewatering. Additionally, trans-zeatin/ABA and isopentenyladenine/ABA ratios, which were decreased in water-stressed plants, recovered rapidly in rewatered plants. Spraying plants with 6-benzylaminopurine (0.1-100 µM) did not influence recovery of either stomatal conductance or needle water status. It can be concluded that the delayed recovery of stomatal conductance in Scots pine needles was not due to sustained ABA accumulation or a sustained decrease in the CK/ABA ratio, and CK supplementation was unable to overcome this delayed recovery.

Effect of Abscisic Acid on Growth, Fatty Acid Profile, and Pigment Composition of the Chlorophyte Chlorella (Chromochloris) zofingiensis and Its Co-Culture Microbiome.

Microalga Chlorella (Chromochloris) zofingiensis has been gaining increasing attention of investigators as a potential competitor to Haematococcus pluvialis for astaxanthin and other xanthophylls production. Phytohormones, including abscisic acid (ABA), at concentrations relevant to that in hydroponic wastewater, have proven themselves as strong inductors of microalgae biomass productivity and biosynthesis of valuable molecules. The main goal of this research was to evaluate the influence of phytohormone ABA on the physiology of C. zofingiensis in a non-aseptic batch experiment. Exogenous ABA stimulated C. zofingiensis cell division, biomass production, as well as chlorophyll, carotenoid, and lipid biosynthesis. The relationship between exogenous ABA concentration and the magnitude of the observed effects was non-linear, with the exception of cell growth and biomass production. Fatty acid accumulation and composition depended on the concentration of ABA tested. Exogenous ABA induced spectacular changes in the major components of the culture microbiome of C. zofingiensis. Thus, the abundance of the representatives of the genus Rhodococcus increased drastically with an increase in ABA concentration, whereas the abundance of the representatives of Reyranella and Bradyrhizobium genera declined. The possibilities of exogenous ABA applications for the enhancing of the biomass, carotenoid, and fatty acid productivity of the C. zofingiensis cultures are discussed.

Manganese Deficiency Suppresses Growth and Photosynthetic Processes but Causes an Increase in the Expression of Photosynthetic Genes in Scots Pine Seedlings.

Manganese deficiency is a serious plant nutritional disorder, resulting in the loss of crop productivity in many parts of the world. Despite the progress made in the study of angiosperms, the demand for Mn in gymnosperms and the physiological responses to Mn deficiency remain unexplored. We studied the influence of Mn deficiency for 24 weeks on Pinus sylvestris L. seedling growth, ion homeostasis, pigment contents, lipid peroxidation, chlorophyll fluorescence indices and the transcript levels of photosynthetic genes and genes involved in chlorophyll biosynthesis. It was shown that Mn-deficient plants demonstrated suppressed growth when the Mn content in the needles decreased below 0.34 µmol/g DW. The contents of photosynthetic pigments decreased when the Mn content in the needles reached 0.10 µmol/g DW. Mn deficiency per se did not lead to a decrease in the nutrient content in the organs of seedlings. Photoinhibition of PSII was observed in Mn-deficient plants, although this was not accompanied by the development of oxidative stress. Mn-deficient plants had an increased transcript abundance of genes (psbO, psbP, psbQ, psbA and psbC), encoding proteins directly associated with the Mn cluster also as other proteins involved in photosynthesis, whose activities do not depend on Mn directly. Furthermore, the transcript levels of the genes encoding the large subunit of Rubisco, light-dependent NADPH-protochlorophyllide oxidoreductase and subunits of light-independent protochlorophyllide reductase were also increased in Mn-deficient plants.

Stem notching decreases stem hydraulic conductance but does not influence drought impacts and post-drought recovery in Scots pine and Norway spruce.

The tight connection between the deterioration of xylem function and plant mortality under drought is well recognized. However, a lack of mechanistic understanding of how substantial conductivity loss influences plant performance under drought and during post-drought recovery hinders our ability to model tree responses to drought stress. We artificially induced a loss of 50% of xylem conducting area in Scots pine and Norway spruce saplings by stem notching and investigated plant performance under drought and during post-drought recovery. Plant mortality, xylem hydraulic conductivity, leaf water status and stomatal conductance were measured. We observed no preferential mortality of top plant parts (above the notches) compared to basal plant parts (below the notches), and no consistent trend in hydraulic conductivity loss was observed between top and basal parts of dying plants. Stem hydraulic conductivity, water status of the needles and stomatal conductance changed similarly between the top and basal parts during drought and post-drought recovery, which indicated the substantial hydraulic overcapacity of the stems. The recovery of stomatal conductance demonstrated prominent hysteresis due to non-hydraulic stomatal limitations. The results obtained are highly important for modelling the influence of plant hydraulic impairment on plant performance under drought and during post-drought recovery.

Mineral Nutrition of Naturally Growing Scots Pine and Norway Spruce under Limited Water Supply.

The deterioration of plant mineral nutrition during drought is a significant factor in the negative influence of drought on plant performance. We aimed to study the effects of seasonal and multiyear water shortages on nutrient supply and demand in Scots pine and Norway spruce. We studied pine and spruce trees naturally grown in the Bryansk region (Russia). The dynamics of several nutrients (K, Ca, Mg, P, Fe, Mn, Zn, and Ca) in wood, needles, and bark of current-year twigs and the dynamics of the available pools of these elements at different soil depths were analysed. To assess the physiological consequences of changes in element concentrations, lipid peroxidation products and photosynthetic pigments were measured in the needles. Water shortage increased the wood concentrations of all elements except for Mn. In pine, this increase was mainly due to seasonal water deficit, whereas in spruce, multiyear differences in water supply were more important. This increased availability of nutrients was not observed in soil-based analyses. In needles, quite similar patterns of changes were found between species, with Mg increasing almost twofold and Fe and Mn decreasing under water shortage, whereas the remainder of the elements did not change much under differing water supplies. Neither the concentrations of photosynthetic pigments nor the contents of lipid peroxidation products correlated with element dynamics in needles. In summary, water shortage increased the availability of all elements except Mn for the plant; however, needle element contents were regulated independently of element availability for plants.

Quantitative analysis of differential dehydrin regulation in pine and spruce seedlings under water deficit.

Dehydrins are well-known components of plant responses to different stresses that cause dehydration, including drought, freezing, salinity, etc. In conifers, the dehydrin gene family is very large, implying that the members of this family have important physiological functions in conifer stress tolerance. However, dehydrin gene expression displays a wide range of responses to stress, from thousand-fold increased expression to decreased expression, and it is generally unknown how regulatory systems are connected at the mRNA and protein levels. Therefore, we studied these aspects of dehydrin regulation in Scots pine (Pinus sylvestris L.) and Norway spruce (Picea abies (L.) H. Karst) seedlings under polyethylene glycol 6000-induced osmotic stress ranging from relatively low (culture medium water potential of -0.15 MPa) to very high (-1.0 MPa) intensities. In pine, the major dehydrin protein was Dhn1 in both the roots and needles, and in spruce, two isoforms of the Dhn4 protein were the major dehydrins; both of these proteins are AESK-type dehydrins. The genes encoding these major proteins were highly expressed even under control conditions; surprisingly, we also observed several highly expressed dehydrin genes that were not abundantly translated. Under osmotic stress, the most prominent expression changes were observed for the dehydrin genes with low basal expression levels, whereas highly expressed genes generally demonstrated rather modest changes in expression. We report proposed constitutive physiological functions of the AESK-type dehydrins in Pinaceae plants.

Phytotoxicity of short-term exposure to excess zinc or copper in Scots pine seedlings in relation to growth, water status, nutrient balance, and antioxidative activity.

The toxic effects of heavy metals pose a significant threat to the productivity and stability of forest ecosystems. Changes in the agrochemical properties of polluted forest soils due to global climate changes can increase the bioavailability of previously immobilized heavy metals. To test this hypothesis, we studied the effects of short-term shock exposure to ZnSO4 (50, 150, 300 µM) or CuSO4 (2.5, 5, 10 µM) in hydroculture on 4- to 6-week-old seedlings of Scots pine (Pinus sylvestris L.) with well-developed root systems. The effects of the excess heavy metals on mineral nutrients and the functioning of low-molecular-weight antioxidants and glutathione in protecting plants from oxidative damage were studied. Even short-term exposure to exogenous metals led to their rapid accumulation in the root system and their subsequent transport to aboveground organs. An increase in the 4-hydroxyalkenals content in seedling needles exposed to excess Cu led to an increase in the content of proanthocyanidins and catechins, which act as scavengers of reactive oxygen species. The impact of both metals led to the rapid development of mineral nutrient imbalances in the seedlings, which were most pronounced in the presence of excess Zn. Exposure to excess Zn led to a disruption in the translocation of Fe and a decrease in the Fe content in the needles. The most dramatic consequence of Zn exposure was the development of Mn deficiency in the roots, which was the likely cause of the inhibition of phenolic compound synthesis. A deficiency in phenolic compounds can have serious environmental consequences for pine populations that are at risk of contamination by Zn and Cu salts.

Profiles of endogenous phytohormones and expression of some hormone-related genes in Scots pine and Norway spruce seedlings under water deficit.

Different plant hormones are involved in plant adaptation to water deficit. In comparison to angiosperms, little is known about the impact of drought on the pool of phytohormones in gymnosperms. Therefore, we studied the effect of polyethylene glycol-induced water deficit on the changes in content of different phytohormones in Scots pine and Norway spruce seedlings, which are known for their different strategies of adaptation to water deficit. The following hormone classes were analysed: cytokinins, auxins, jasmonates, salicylic and benzoic acids, and 1-aminocyclopropane-1-carboxylic acid (an ethylene precursor). No consistent reaction to water stress was observed for the content of well-known stress-related hormones - salicylic acid and jasmonates. In contrast, drought induced a dose-dependent accumulation of cytokinins in pine needles, with less profound changes in spruce needles. The most prominent changes were observed for 1-aminocyclopropane-1-carboxylic acid content, which increased several-fold in spruce roots and pine needles under water deficit. Water-deficit-induced changes in the contents of cytokinins and 1-aminocyclopropane-1-carboxylic acid were accompanied by the differential regulation of genes involved in the metabolism of these hormones. Possible links between changes in hormone pools and the adaptation of seedlings to water deficit are discussed.

Specificity of Cd, Cu, and Fe effects on barley growth, metal contents in leaves and chloroplasts, and activities of photosystem I and photosystem II.

Cd, Cu, and Fe were used to reveal the specificity of their toxic actions. We studied the effects of heavy metals on the growth of barley seedlings, contents of cations in leaves and chloroplasts, induced chlorophyll fluorescence and P700 light absorption. Differences were found at each level of research. We measured the contents of Cd, Cu, Fe, Mn, Zn, Ca, Mg, and K. The proportion of cations in leaves targeted to chloroplasts varied from 0.1% (K) to >90% (Fe). Their levels changed in different ways. We found no correlation between changes in cation contents in leaves and chloroplasts. Treatment with Cd, Cu, and Fe increased the contents of some cations. The extra portions were targeted primarily out of chloroplasts, which was most noticeable in the case of Cu and Fe. Cd treatment decreased non-photochemical quenching with concomitant increases in closed photosystem II. We introduced new coefficients qC for closed photosystem II and X(II) to compare the yields of photosystem II and photosystem I. Cd likely decreased both PSI content in leaves and its quantum yield. In control plants, the quantum yield ratio of PSI/PSII increased gradually from 1.25 under low light to 4 under high light. Cd treatment prevented the increase under moderate light; under high light the ratio reached 2. Cu treatment increased the acceptor side limitation of photosystem I under low light; components of the Calvin cycle likely demand more light for activation in Cu-treated plants.

Comparative photosynthetic responses of Norway spruce and Scots pine seedlings to prolonged water deficiency.

Stressors of different natures, including drought stress, substantially compromise the ability of plants to effectively and safely utilize light energy. We investigated the influence of water stress on the photosynthetic processes in Picea abies and Pinus sylvestris, two species with contrasting drought sensitivities. Spruce and pine seedlings were exposed to polyethylene glycol 6000-induced water deficits of different intensities and durations. The maintenance of photosystem I (PSI) oxidation in spruce required increased photosynthetic control and led to the increased reduction of the plastoquinone pool, which was not the case in pine seedlings. As a result of increased excitation pressure, photosystem II (PSII) inactivation was observed in spruce plants, whereas in pine, the decreased PSII photochemistry was likely due to sustained non-photochemical quenching. Downregulation of PSII photochemistry and maintenance of PSI in an oxidized state were linked with the prevention of oxidative stress, even under severe water deficit. The decreased photosynthetic pigment content and photosynthetic gene expression suggested the coordinated downregulation of photosynthetic apparatus components under water stress to reduce light energy absorption. In summary, the observed adaptative mechanisms of pine and spruce to water stress may be similar to the well-studied adaptative mechanisms to winter stress, which may indicate the universality of protective mechanisms under various stresses in conifers.

The labile zinc pool in plant cells.

Zinc is the most abundant and important transition metal in plants; however, the dynamic aspects of zinc homeostasis in plant cells are poorly understood. In this study we explored the pool of labile exchangeable zinc complexes in plant cells, and the potential influence of changes in intracellular zinc availability on cellular physiology. Work was performed on cultivated cell extracts of Arabidopsis thaliana (L.) Heynh. and Thellungiella salsuginea (Pall.) O.E. Schulz grown under control (3.48 µM Zn2+), 10-fold Zn excess or Zn starvation conditions. The free and labile Zn contents in the extracts were then determined by fluorimetric titration. We observed for the first time that plant cells contain micromolar concentrations of labile zinc complexes that account for a low percentage of the total zinc content. Labile zinc is mainly protein bound. Zn starvation inhibits cell proliferation and leads to the disappearance of the labile zinc pool, whereas Zn excess drastically increases the labile zinc pool. Free Zn2+ is buffered at picomolar concentrations in the intracellular milieu, and the increase in free Zn2+ concentrations to low nanomolar values clearly modulates enzyme activity by direct reversible binding. Such increases in free Zn2+ can be achieved by the substantial influx of additional zinc or by the oxidation of zinc-binding thiols. The observed features of the labile zinc pool in plant cells suggest it has a role in intracellular zinc trafficking and zinc signalling.

Comparative analysis of abscisic acid levels and expression of abscisic acid-related genes in Scots pine and Norway spruce seedlings under water deficit.

Abscisic acid (ABA) is one of the main participants in the regulation of plant responses to water deficiency. Knowledge of the ABA signal transduction pathways in gymnosperms is rather limited, especially in comparison with those in angiosperms. Seedlings of Scots pine and Norway spruce are known for their contrasting behaviour strategies under water deficit. To characterize the possible role of ABA in these differences, ABA dynamics were investigated under conditions of water deficit in seedlings of these two species. The content of ABA and its catabolites was followed in the roots and needles of seedlings of Pinus sylvestris and Picea abies under conditions of polyethylene glycol (PEG)-induced water deficiency (-0.15 and -0.5 MPa) for 10 days. The expression of the main genes for ABA-biosynthetic enzymes was also analysed. ABA showed more pronounced stress-dependent dynamics in pine roots than in spruce roots, whereas in needles, the response was greater for spruce than pine. The ABA increase during drought was mainly due to de novo synthesis and the shift in the balance between ABA synthesis and catabolism towards synthesis. The ABA-glucosyl ester did not serve as a reserve for the release of free ABA under water deficiency. The expression levels of the main ABA biosynthetic genes showed a weak or no correlation with changes in ABA content under water stress, i.e., the ABA content in the seedlings of both species was not directly linked to the transcript levels of the main ABA biosynthetic genes. Less-pronounced stress-induced changes in ABA in pine needles than in spruce needles may be related to pine seedlings having a less conservative strategy of growth and maintenance of water balance under water deficit.

Impact of weak water deficit on growth, photosynthetic primary processes and storage processes in pine and spruce seedlings.

We investigated the influence of 40 days of drought on growth, storage processes and primary photosynthetic processes in 3-month-old Scots pine and Norway spruce seedlings growing in perlite culture. Water stress significantly affected seedling water status, whereas absolute dry biomass growth was not substantially influenced. Water stress induced an increase in non-structural carbohydrate content (sugars, sugar alcohols, starch) in the aboveground part of pine seedlings in contrast to spruce seedlings. Due to the relatively low content of sugars and sugar alcohols in seedling organs, their expected contribution to osmotic potential changes was quite low. In contrast to biomass accumulation and storage, photosynthetic primary processes were substantially influenced by water shortage. In spruce seedlings, PSII was more sensitive to water stress than PSI. In particular, electron transport in PSI was stable under water stress despite the substantial decrease of electron transport in PSII. The increase in thermal energy dissipation due to enhancement of non-photochemical quenching (NPQ) was evident in both species under water stress. Simultaneously, the yields of non-regulated energy dissipation in PSII were decreased in pine seedlings under drought. A relationship between growth, photosynthetic activities and storage processes is analysed under weak water deficit.

Distribution of Cd and other cations between the stroma and thylakoids: a quantitative approach to the search for Cd targets in chloroplasts.

Plant growth and photosynthetic activity are usually inhibited due to the overall action of Cd on a whole organism, though few cadmium cations can invade chloroplasts in vivo. We found that in vivo, the major portion of Cd in barley chloroplasts is located in the thylakoids (80%), and the minor portion is in the stroma (20%). Therefore, the electron-transport chain in the thylakoids would be the likely target for direct Cd action in vivo. In vitro, we found the distribution of Cd to be shifted to the stroma (40-60%). In barley chloroplasts, the major portions of Mg, Fe, Mn, and Cu were found to be located in the thylakoids, and most Ca, Zn, and K in the stroma. This finding was true for both control and Cu- or Fe-treated plants. Treatment with Cd affected the contents of all cations, and the largest portions of Ca and Zn were in the thylakoids. Alterations of the K and Mn contents were caused by Cd, Cu, or Fe treatment; the levels of other cations in chloroplasts were changed specifically by Cd treatment. The quantity of Cd in chloroplasts was small in comparison to that of Mg, Ca, and Fe. In thylakoids, the amount of Cd was similar to that of Cu and comparable to the levels of Zn and Mn. Accordingly, the possible targets for direct Cd action in thylakoids are the Mn cluster, plastocyanin, carbonic anhydrase, or FtsH protease. The quantity of Cd in thylakoids is sufficient to replace a cation nearly completely at one of these sites or partially (20-30%) at many of these sites.

Long-term impact of cement plant emissions on the elemental composition of both soils and pine stands and on the formation of Scots pine seeds.

We investigated the long-term impact of the largest Russian cement plant on mesopodzol sandy soils and Scots pine stands. We determined the distributions of the total and available pools of Ca, Mg, K, Na, Mn, Fe, Zn, Ni, Cu, Pb and Cd in the soil profile to a depth of 60 cm (illuvial horizon) as well as the accumulation patterns of these elements in the vegetative and generative organs of Scots pine trees. High Ca accumulation in the impact zone soils was a result of CaO emissions by a cement plant. Also, CaO became the main cause of soil profile alkalization due to neutralization of soil acids and formation of calcium hydroxide or carbonates. Alkalization immobilized substantial amounts of Fe, Mn, Zn and Ni in the soil, reducing their availability. The most prominent effect of long-term cement production was a prominent Mn deficiency in vegetative and generative Scots pine organs due to the exhaustion of the available Mn pool in the illuvial horizon. The miniaturization of cones, a decrease in seed yield and a reduction in seed germinability were observed in the emission impact zones. Pretreatment of Mn-deficient seeds with manganese eliminated Mn deficiency but did not increase seed germination.

Impact of drought stress induced by polyethylene glycol on growth, water relations and cell viability of Norway spruce seedlings.

We investigated physiological responses of 7-week-old Norway spruce seedlings to water deficits of different intensities. Hydroponically grown seedlings were subjected to mild (-0.15 MPa), strong (-0.5 and -1.0 MPa) and extreme (-1.5 MPa) water deficit induced by polyethylene glycol 6000, and their growth parameters, water status and physiological activity were analyzed. Seedlings effectively restricted water loss under drought, and even under extreme water deficit, shoot relative water content did not fall below 85%. Water stress induced substantial decreases in the osmotic potentials of root and needle cell sap, up to 0.3-0.4 MPa under extreme water deficit, though this did not result from water loss or accumulation of K+ and Na+ ions. Seedling growth was very susceptible to water stress because of poor capacity for cell wall adjustment. Water stress injured seedling roots, as evidenced by the loss of root cell physiological activity estimated by the ability to hydrolyse fluorescein diacetate and by increased root calcium content up to 8-10-fold under extreme water stress. At the same time, root hair growth was enhanced, especially under mild water deficit, which increased the root water-absorbing capacity. In summary, seedlings of Norway spruce were characterized by high susceptibility to water stress and concurrently by pronounced ability to maintain water status. These characteristics are fully consistent with spruce confinement to moist habitats.

Different roles of glutathione in copper and zinc chelation in Brassica napus roots.

We investigated the specific features of copper and zinc excess action on the roots of canola (Brassica napus L.) plants. Copper rapidly accumulated in canola root cells and reached saturation during several hours of treatment, whereas the root zinc content increased relatively slowly. Excessive copper and zinc entry inside the cell resulted in significant cell damage, as evidenced by alterations in plasmalemma permeability and decreases in cellular enzymatic activity. Zinc excess specifically damaged root hair cells, which correlated with a pronounced elevation of their labile zinc level. In vitro, we showed that reduced glutathione (GSH) readily reacted with copper ions to form complexes with blocked sulfhydryl groups. In contrast, zinc ions were ineffective as glutathione blockers, and glutathione molecules did not lose their specific chemical activity in the presence of Zn2+ ions. The effect of copper and zinc excess on the glutathione pool in canola root cells was analysed by a combination of biochemical determination of total and oxidized glutathione contents and fluorescent staining of free reduced glutathione with monochlorobimane dye. Excess copper led to dose-dependent diminution of free reduced glutathione contents in the root cells, which could not be explained by the loss of total cellular glutathione or its oxidation. In contrast, we observed little effect of much higher intracellular zinc concentrations on the free reduced glutathione content. We concluded that GSH plays an important role in copper excess, but not zinc excess chelation, in canola root cells.

Effects of copper deficiency and copper toxicity on organogenesis and some physiological and biochemical responses of Scots pine (Pinus sylvestris L.) seedlings grown in hydroculture.

The morphological, physiological, and biochemical parameters of 6-week-old seedlings of Scots pine (Pinus sylvestris L.) were studied under deficiency (1.2 nM) and chronic exposure to copper (0.32, 1, 2.5, 5, and 10 µM CuSO4) in hydroculture. The deposit of copper in the seed allowed the seedlings to develop under copper deficiency without visible disruption of growth. The high sensitivity of Scots pine to the toxic effects of copper was shown, which manifested as a significant inhibition of growth and development. The loss of dominance of the main root and a strong inhibition of lateral root development pointed to a lack of adaptive reorganization of the root system architecture under copper excess. A preferential accumulation of copper in the root and a minor translocation in aerial organs confirmed that Scots pine belongs to a group of plants that exclude copper. Selective impairment in the absorption of manganese was discovered, under both deficiency and excess of copper in the nutrient solution, which was independent of the degree of development of the root system. Following 10 µM CuSO4 exposure, the absorption of manganese and iron from the nutrient solution was completely suppressed, and the development of seedlings was secured by the stock of these micronutrients in the seed. The absence of signs of oxidative stress in the seedling organs was shown under deficiency and excess of copper, as evidenced by the steady content of malondialdehyde and 4-hydroxyalkenals. Against this background, no changes in total superoxide dismutase activity in the organs of seedlings were revealed, and the increased content of low-molecular-weight antioxidants was observed in the roots under 1 µM and in the needles under 5 µM CuSO4 exposures.

Blue light alters miR167 expression and microRNA-targeted auxin response factor genes in Arabidopsis thaliana plants.

The effect of blue LED (450 nm) on the photomorphogenesis of Arabidopsis thaliana Col-0 plants and the transcript levels of several genes, including miRNAs, photoreceptors and auxin response factors (ARF) was investigated. It was observed that blue light accelerated the generative development, reduced the rosette leaf number, significantly reduced the leaf area, dry biomass and led to the disruption of conductive tissue formation. The blue LED differentially influenced the transcript levels of several phytochromes (PHY a, b, c, d, and e), cryptochromes (CRY 1 and 2) and phototropins (PHOT 1 and 2). At the same time, the blue LED significantly increased miR167 expression compared to a fluorescent lamp or white LEDs. This increase likely resulted in the enhanced transcription of the auxin response factor genes ARF4 and ARF8, which are regulated by this miRNA. These findings support the hypothesis that the effects of blue light on A. thaliana are mediated by auxin signalling pathway involving miRNA-dependent regulation of ARF gene expression.