RESUMO
Winter crops acquire frost tolerance during the process of cold acclimation when plants are exposed to low but non-freezing temperatures that is connected to specific metabolic adjustments. Warm breaks during/after cold acclimation disturb the natural process of acclimation, thereby decreasing frost tolerance and can even result in a resumption of growth. This phenomenon is called deacclimation. In the last few years, studies that are devoted to deacclimation have become more important (due to climate changes) and necessary to be able to understand the mechanisms that occur during this phenomenon. In the acclimation of plants to low temperatures, the importance of plant membranes is indisputable; that is why the main aim of our studies was to answer the question of whether (and to what extent) deacclimation alters the physicochemical properties of the plant membranes. The studies were focused on chloroplast membranes from non-acclimated, cold-acclimated and deacclimated cultivars of winter oilseed rape. The analysis of the membranes (formed from chloroplast lipid fractions) using the Langmuir technique revealed that cold acclimation increased membrane fluidity (expressed as the Alim values), while deacclimation generally decreased the values that were induced by cold. Moreover, because the chloroplast membranes were penetrated by lipophilic molecules such as carotenoids or tocopherols, the relationships between the structure of the lipids and the content of these antioxidants in the chloroplast membranes during the process of the cold acclimation and deacclimation of oilseed rape are discussed.
Assuntos
Aclimatação , Cloroplastos , Temperatura Baixa , Aclimatação/fisiologia , Cloroplastos/metabolismo , Brassica napus/metabolismo , Brassica napus/fisiologia , Carotenoides/metabolismo , Fluidez de Membrana/fisiologia , Membranas Intracelulares/metabolismoRESUMO
Winter plants acclimate to frost mainly during the autumn months, through the process of cold acclimation. Global climate change is causing changes in weather patterns such as the occurrence of warmer periods during late autumn or in winter. An increase in temperature after cold acclimation can decrease frost tolerance, which is particularly dangerous for winter crops. The aim of this study was to investigate the role of brassinosteroids (BRs) and BR analogues as protective agents against the negative results of deacclimation. Plants were cold-acclimated (3 weeks, 4 °C) and deacclimated (1 week, 16/9 °C d/n). Deacclimation generally reversed the cold-induced changes in the level of the putative brassinosteroid receptor protein (BRI1), the expression of BR-induced COR, and the expression of SERK1, which is involved in BR signal transduction. The deacclimation-induced decrease in frost tolerance in oilseed rape could to some extent be limited by applying steroid regulators. The deacclimation in plants could be detected using non-invasive measurements such as leaf reflectance, chlorophyll a fluorescence, and gas exchange monitoring.
Assuntos
Aclimatação , Brassica napus , Brassinosteroides , Temperatura Baixa , Regulação da Expressão Gênica de Plantas , Brassinosteroides/metabolismo , Brassica napus/fisiologia , Brassica napus/metabolismo , Estações do Ano , Proteínas de Plantas/metabolismo , Folhas de Planta/metabolismo , Folhas de Planta/fisiologiaRESUMO
High temperatures associated with climate change may increase the severity of plant diseases. This study investigated the effect of heat shock treatment on host and non-host barley powdery mildew interactions using brassinosteroid (BR) mutants of barley. Brassinosteroids are plant steroid hormones, but so far little is known about their role in plant-fungal interactions. Wild type barley cultivar Bowman and its near-isogenic lines with disturbances in BR biosynthesis or signalling showed high compatibility to barley powdery mildew race A6, while cultivar Delisa and its BR-deficient mutants 522DK and 527DK were fully incompatible with this pathogen (host plant-pathogen interactions). On the other hand, Bowman and its mutants were highly resistant to wheat powdery mildew, representing non-host plant-pathogen interactions. Heat pre-treatment induced shifts in these plant-pathogen interactions towards higher susceptibility. In agreement with the more severe disease symptoms, light microscopy showed a decrease in papillae formation and hypersensitive response, characteristic of incompatible interactions, when heat pre-treatment was applied. Mutant 527DK, but not 522DK, maintained high resistance to barley powdery mildew race A6 despite heat pre-treatment. By 10 days after heat treatment and infection, a noticeable shift became apparent in the chlorophyll a fluorescence and in various leaf reflectance parameters at all genotypes.
RESUMO
Nitric oxide (NO) is a signaling molecule that participates in plant adaptation to adverse environmental factors. This study aimed to clarify the role of NO in the regulation of vacuolar H+-ATPase (V-ATPase) in the roots of cucumber seedlings grown under control and Cd stress conditions. In addition, the relationship between NO and salicylic acid (SA), as well as their interrelations with hydrogen sulfide (H2S) and hydrogen peroxide (H2O2), have been verified. The effect of NO on V-ATPase was studied by analyzing two enzyme activities, the expression level of selected VHA genes and the protein level of selected VHA subunits in plants treated with a NO donor (sodium nitroprusside, SNP) and NO biosynthesis inhibitors (tungstate, WO42- and N-nitro-L-arginine methyl ester, L-NAME). Our results indicate that NO functions as a positive regulator of V-ATPase and that this regulation depends on NO generated by nitrate reductase and NOS-like activity. It was found that the mechanism of NO action is not related to changes in the gene expression or protein level of the V-ATPase subunits. The results suggest that in cucumber roots, NO signaling interacts with the SA pathway and, to a lesser extent, with two other known V-ATPase regulators, H2O2 and H2S.
RESUMO
BACKGROUND: In earlier studies [1], we indicated that applying brassinosteroids (BRs) to lipids that had been isolated from plants altered the physicochemical properties of the monolayers. A continuation of these dependencies using the defined model lipid systems is presented in this paper. The influence of homocastasterone (HCS) and castasterone (CS) (BRs for which the increase in concentration were characteristic of plants grown at low temperatures) on the membrane properties of their polar and the hydrophobic parts were studied. RESULTS: Changes in the electrokinetic potential indicate that both BRs decreased the negative charge of the surface, which is an important factor in modifying the contacts with the polar substances. This property of BRs has not yet been described. The studies of the interactions that occur in the hydrophobic part of the membrane were investigated using the EPR methods and Langmuir techniques. The physicochemical parameters of the lipid structure were determined, and the excess of Gibbs free energy was calculated. CONCLUSION: We conclude that examined BRs modify both the hydrophilic and hydrophobic properties of the membranes, but to a greater extent HCS. The consequence of these changes may be the attempt to maintain the stability of the membranes in stressful temperature conditions and / or to the possibility of adsorption of other substances on membranes surfaces. The change of plant metabolism towards increasing the amount of BR, mainly HCS (under cooling) may by an important factor for maintaining optimal structural properties of membranes and their functionality despite temperature changes.
Assuntos
Brassinosteroides , Temperatura Baixa , Brassinosteroides/metabolismo , TemperaturaRESUMO
The objective of this study was to answer the question of how the deacclimation process affects frost tolerance, photosynthetic efficiency, brassinosteroid (BR) homeostasis and BRI1 expression of winter oilseed rape. A comparative study was conducted on cultivars with different agronomic and physiological traits. The deacclimation process can occur when there are periods of higher temperatures, particularly in the late autumn or winter. This interrupts the process of the acclimation (hardening) of winter crops to low temperatures, thus reducing their frost tolerance and becoming a serious problem for agriculture. The experimental model included plants that were non-acclimated, cold acclimated (at 4 °C) and deacclimated (at 16 °C/9 °C, one week). We found that deacclimation tolerance (maintaining a high frost tolerance despite warm deacclimating periods) was a cultivar-dependent trait. Some of the cultivars developed a high frost tolerance after cold acclimation and maintained it after deacclimation. However, there were also cultivars that had a high frost tolerance after cold acclimation but lost some of it after deacclimation (the cultivars that were more susceptible to deacclimation). Deacclimation reversed the changes in the photosystem efficiency that had been induced by cold acclimation, and therefore, measuring the different signals associated with photosynthetic efficiency (based on prompt and delayed chlorophyll fluorescence) of plants could be a sensitive tool for monitoring the deacclimation process (and possible changes in frost tolerance) in oilseed rape. Higher levels of BR were characteristic of the better frost-tolerant cultivars in both the cold-acclimated and deacclimated plants. The relative expression of the BRI1 transcript (encoding the BR-receptor protein) was lower after cold acclimation and remained low in the more frost-tolerant cultivars after deacclimation. The role of brassinosteroids in oilseed rape acclimation/deacclimation is briefly discussed.
Assuntos
Brassica napus , Aclimatação/fisiologia , Brassica napus/genética , Brassinosteroides , Temperatura Baixa , Homeostase , FotossínteseRESUMO
Although the only known steroid hormones in plants are brassinosteroids, interestingly, mammalian steroid hormones such as androgens or estrogens are also part of the plant metabolic profile. This presented review is focused on the progress that has been made in this matter during the last two decades. The presence of testosterone, 17ß-estradiol, and other androgens/estrogens in plants (particularly those that can be measured using more advanced techniques) is described. The physiological activity of androgens and estrogens, especially in plants' stress response, are discussed, together with some possible mechanisms of their action. The current knowledge indicates that although androgens and estrogens do not have the status of hormones in plants, they are physiologically active and can serve as regulators that support the activity of classic hormones in (1) regulating the various processes connected with plant growth and development and (2) the interaction of plants with their environment.
RESUMO
The objective of our study was to characterise the growth of tomato seedlings under various light spectra, but special attention has been paid to gaining a deeper insight into the details of photosynthetic light reactions. The following light combinations (generated by LEDs, constant light intensity at 300 µmol m-2 s-1) were used: blue/red light; blue/red light + far red; blue/red light + UV; white light that was supplemented with green, and white light that was supplemented with blue. Moreover, two combinations of white light for which the light intensity was changed by imitating the sunrise, sunset, and moon were also tested. The reference point was also light generated by high pressure sodium lamps (HPS). Plant growth/morphological parameters under various light conditions were only partly correlated with the photosynthetic efficiency of PSI and PSII. Illumination with blue/red as the main components had a negative effect on the functioning of PSII compared to the white light and HPS-generated light. On the other hand, the functioning of PSI was especially negatively affected under the blue/red light that was supplemented with FR. The FT-Raman studies showed that the general metabolic profile of the leaves (especially proteins and ß-carotene) was similar in the plants that were grown under the HPS and under the LED-generated white light for which the light intensity changed during a day. The effect of various light conditions on the leaf hormonal balance (auxins, brassinosteroids) is also discussed.
Assuntos
Fotossíntese , Solanum lycopersicum/metabolismo , Solanum lycopersicum/efeitos da radiação , Brassinosteroides/metabolismo , Clorofila/metabolismo , Ácidos Indolacéticos/metabolismo , Luz , Solanum lycopersicum/crescimento & desenvolvimento , Metaboloma , Fotossíntese/efeitos da radiação , Complexo de Proteína do Fotossistema I/metabolismo , Complexo de Proteína do Fotossistema I/efeitos da radiação , Complexo de Proteína do Fotossistema II/metabolismo , Complexo de Proteína do Fotossistema II/efeitos da radiação , Reguladores de Crescimento de Plantas/metabolismo , Folhas de Planta/crescimento & desenvolvimento , Folhas de Planta/metabolismo , Folhas de Planta/efeitos da radiação , Plântula/crescimento & desenvolvimento , Plântula/metabolismo , Plântula/efeitos da radiação , Análise Espectral RamanRESUMO
The aim of the study was to examine combinations of base oils and herbal additives with a view to obtaining macerates with improved health benefits. Base oils were cold-pressed from the seeds of black cumin, borage, evening primrose, safflower, walnut, common hazel, and oilseed rape, as well as the flesh of sea-buckthorn fruits. They were then supplemented with herbs, including basil, thyme, and sage, in order to create macerates. Total antioxidant activity and tocopherol level were analyzed in oils, macerates, and oil cakes. Additionally, chemical properties of oil cakes-such as the level of fibre, vitamin C, ß-carotene, and lutein-were also examined. Supplementation with herbs caused diversified effects on antioxidant activity and tocopherol level in macerates depending on the base oil, herb, and supplementation method. The obtained results indicate that tocopherol level does not play a decisive role in determining the antioxidant properties of oils, macerates, and oil cakes, suggesting significant involvement of other antioxidants. Among the tested macerates, the most promising one seems to be oilseed rape oil enriched with sage or basil to maximize its health benefits. The study can serve as a starting point for the development and implementation of functional macerates and oil cakes in healthy nutrition.
RESUMO
Ecdysteroids (ECs) are steroid hormones originally found in the animal kingdom where they function as insect molting hormones. Interestingly, a relatively high number of these substances can also be formed in plant cells. Moreover, ECs have certain regulatory effects on plant physiology, but their role in plants still requires further study. One of the main aims of the present study was to verify a hypothesis that fenarimol, an inhibitor of the biosynthesis of ECs in the animal kingdom, also affects the content of endogenous ECs in plants using winter wheat Triticum aestivum L. as a model plant. The levels of endogenous ECs in winter wheat, including the estimation of their changes during a course of different temperature treatments, have been determined using a sensitive analytical method based on UHPLC-MS/MS. Under our experimental conditions, four substances of EC character were detected in the tissue of interest in amounts ranging from less than 1 to over 200 pg·g-1 FW: 20-hydroxyecdysone, polypodine B, turkesterone, and isovitexirone. Among them, turkesterone was observed to be the most abundant EC and accumulated mainly in the crowns and leaves of wheat. Importantly, the level of ECs was observed to be dependent on the age of the plants, as well as on growth conditions (especially temperature). Fenarimol, an inhibitor of a cytochrome P450 monooxygenase, was shown to significantly decrease the level of naturally occurring ECs in experimental plants, which may indicate its potential use in studies related to the biosynthesis and physiological function of these substances in plants.
Assuntos
Produtos Biológicos/metabolismo , Ecdisteroides/biossíntese , Pirimidinas/farmacologia , Triticum/metabolismo , Produtos Biológicos/química , Cromatografia Líquida/métodos , Ecdisteroides/química , Fungicidas Industriais/farmacologia , Estrutura Molecular , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/metabolismo , Espectrometria de Massas em Tandem/métodos , Temperatura , Triticum/crescimento & desenvolvimentoRESUMO
Cereals, which belong to the Poaceae family, are the most economically important group of plants. Among abiotic stresses, temperature stresses are a serious and at the same time unpredictable problem for plant production. Both frost (in the case of winter cereals) and high temperatures in summer (especially combined with a water deficit in the soil) can result in significant yield losses. Plants have developed various adaptive mechanisms that have enabled them to survive periods of extreme temperatures. The processes of acclimation to low and high temperatures are controlled, among others, by phytohormones. The current review is devoted to the role of brassinosteroids (BR) in cereal acclimation to temperature stress with special attention being paid to the impact of BR on photosynthesis and the membrane properties. In cereals, the exogenous application of BR increases frost tolerance (winter rye, winter wheat), tolerance to cold (maize) and tolerance to a high temperature (rice). Disturbances in BR biosynthesis and signaling are accompanied by a decrease in frost tolerance but unexpectedly an improvement of tolerance to high temperature (barley). BR exogenous treatment increases the efficiency of the photosynthetic light reactions under various temperature conditions (winter rye, barley, rice), but interestingly, BR mutants with disturbances in BR biosynthesis are also characterized by an increased efficiency of PSII (barley). BR regulate the sugar metabolism including an increase in the sugar content, which is of key importance for acclimation, especially to low temperatures (winter rye, barley, maize). BR either participate in the temperature-dependent regulation of fatty acid biosynthesis or control the processes that are responsible for the transport or incorporation of the fatty acids into the membranes, which influences membrane fluidity (and subsequently the tolerance to high/low temperatures) (barley). BR may be one of the players, along with gibberellins or ABA, in acquiring tolerance to temperature stress in cereals (particularly important for the acclimation of cereals to low temperature).
Assuntos
Brassinosteroides/metabolismo , Membrana Celular/fisiologia , Fenômenos Químicos , Grão Comestível/fisiologia , Fotossíntese , Estresse Fisiológico , Temperatura , Brassinosteroides/químicaRESUMO
The roles of endogenous brassinosteroids (BRs) in the modulation of reaction to drought and genetic regulation of this process are still obscure. In this study, a multidirectional analysis was performed on semi-dwarf barley (Hordeum vulgare) Near-Isogenic Lines (NILs) and the reference cultivar "Bowman" to get insights into various aspects of metabolic reaction to drought. The NILs are defective in BR biosynthesis or signaling and displayed an enhanced tolerance to drought. The BR metabolism perturbations affected the glucose and fructose accumulation under the control and stress conditions. The BR metabolism abnormalities negatively affected the sucrose accumulation as well. However, during drought, the BR-deficient NILs accumulated higher contents of sucrose than the "Bowman" cultivar. Under the control conditions, accumulation of transcripts encoding antioxidant enzymes ascorbate peroxidase (HvAPX) and superoxide dismutase (HvSOD) was BR-dependent. However, during drought, the accumulation of HvAPX transcript was BR-dependent, whereas accumulations of transcripts encoding catalase (HvCAT) and HvSOD were not affected by the BR metabolism perturbations. The obtained results reveal a significant role of BRs in regulation of the HvAPX and HvCAT enzymatic activities under control conditions and the HvAPX and HvSOD activities during physiological reactions to drought.
Assuntos
Brassinosteroides/metabolismo , Hordeum/genética , Hordeum/metabolismo , Ascorbato Peroxidases/genética , Ascorbato Peroxidases/metabolismo , Catalase/genética , Catalase/metabolismo , Secas , Frutose/metabolismo , Genes de Plantas , Glucose/metabolismo , Hordeum/crescimento & desenvolvimento , Peróxido de Hidrogênio/metabolismo , Mutação , Reguladores de Crescimento de Plantas/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Ribulose-Bifosfato Carboxilase/genética , Ribulose-Bifosfato Carboxilase/metabolismo , Estresse Fisiológico/genética , Estresse Fisiológico/fisiologia , Superóxido Dismutase-1/genética , Superóxido Dismutase-1/metabolismoRESUMO
The present work focused on the characterization of some physiological mechanisms activated upon powdery mildew inoculation of the susceptible barley cultivar Ingrid and its near-isogenic lines (NILs) carrying various resistant genes (Mla, Mlg and mlo). After inoculation with Blumeria graminis f. sp. hordei (Bgh), measurements of leaf reflectance and chlorophyll a fluorescence were performed 3 and 7 day post-inoculation (dpi), while hormone assays were made 7 dpi. Bgh-inoculated resistant genotypes were characterized by lowered leaf reflectance parameters that correlated with carotenoids (CRI) and water content (WBI) in comparison to inoculated Ingrid. The PSII activity (i.e., Fv/Fm, ETo/CSm and P.I.ABS) strongly decreased in susceptible Ingrid leaves when the disease symptoms became visible 7 dpi. In Mla plants with visible hypersensitive spots the PSII activity decreased to a lesser extent. Inoculation resulted in a very slight decrease of photosynthesis at later stage of infection in Mlg plants, whereas in resistant mlo plants the PSII activity did not change. Chlorophyll a fluorescence measurements allowed presymptomatic detection of infection in Ingrid and Mla. Changes in the homeostasis of 22 phytohormones (cytokinins, auxins, gibberellins and the stress hormones JA, SA and ABA) in powdery mildew inoculated barley are discussed in relation to resistance against this biotrophic pathogen.
Assuntos
Ascomicetos/patogenicidade , Hordeum/metabolismo , Doenças das Plantas/genética , Ascomicetos/efeitos dos fármacos , Clorofila A/metabolismo , Resistência à Doença/genética , Regulação da Expressão Gênica de Plantas/genética , Hordeum/genética , Fotossíntese/fisiologia , Folhas de Planta/metabolismo , Proteínas de Plantas/metabolismoRESUMO
(1) Background: The study characterized barley mutants with brassinosteroid (BR) biosynthesis and signaling disturbances in terms of the physicochemical/structural properties of membranes to enrich the knowledge about the role of brassinosteroids for lipid metabolism and membrane functioning. (2) Methods: The Langmuir method was used to investigate the properties of the physicochemical membranes. Langmuir monolayers were formed from the lipid fractions isolated from the plants growing at 20 °C and then acclimated at 5 °C or 27 °C. The fatty acid composition of the lipids was estimated using gas chromatography. (3) Results: The BR-biosynthesis and BR-signaling mutants of barley were characterized by a temperature-dependent altered molar percentage of fatty acids (from 14:0 to 20:1) in their galactolipid and phospholipid fractions in comparison to wild-type (WT). For example, the mutants had a lower molar percentage of 18:3 in the phospholipid (PL) fraction. The same regularity was observed at 5 °C. It resulted in altered physicochemical parameters of the membranes (Alim, πcoll, Cs-1). (4) Conclusions: BR may be involved in regulating fatty acid biosynthesis or their transport/incorporation into the cell membranes. Mutants had altered physicochemical parameters of their membranes, compared to the WT, which suggests that BR may have a multidirectional impact on the membrane-dependent physiological processes.
Assuntos
Aclimatação/fisiologia , Membrana Celular/metabolismo , Genes de Plantas , Hordeum/genética , Hordeum/fisiologia , Mutação/genética , Temperatura , Fenômenos Químicos , Ácidos Graxos/metabolismo , Hordeum/crescimento & desenvolvimento , Lipídeos de Membrana/metabolismoRESUMO
In temperature stress, the main role of heat-shock proteins (HSP) is to act as molecular chaperones for other cellular proteins. However, knowledge about the hormonal regulation of the production of the HSP is quite limited. Specifically, little is known about the role of the plant steroid hormones-brassinosteroids (BR)-in regulating the HSP expression. The aim of our study was to answer the question of how a BR deficit or disturbances in its signaling affect the accumulation of the HSP90, HSP70, HSP18, and HSP17 transcripts and protein in barley growing at 20 °C (control) and during the acclimation of plants at 5 °C and 27 °C. In barley, the temperature of plant growth modified the expression of HSPs. Furthermore, the BR-deficient mutants (mutations in the HvDWARF or HvCPD genes) and BR-signaling mutants (mutation in the HvBRI1 gene) were characterized by altered levels of the transcripts and proteins of the HSP group compared to the wild type. The BR-signaling mutant was characterized by a decreased level of the HSP transcripts and heat-shock proteins. In the BR-deficient mutants, there were temperature-dependent cases when the decreased accumulation of the HSP70 and HSP90 transcripts was connected to an increased accumulation of these HSP. The significance of changes in the accumulation of HSPs during acclimation at 27 °C and 5 °C is discussed in the context of the altered tolerance to more extreme temperatures of the studied mutants (i.e., heat stress and frost, respectively).
Assuntos
Brassinosteroides/biossíntese , Proteínas de Choque Térmico/genética , Proteínas de Choque Térmico/metabolismo , Hordeum/crescimento & desenvolvimento , Aclimatação , Vias Biossintéticas , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Hordeum/genética , Hordeum/metabolismo , Mutação , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , TemperaturaRESUMO
Plants have developed various acclimation strategies in order to counteract the negative effects of abiotic stresses (including temperature stress), and biological membranes are important elements in these strategies. Brassinosteroids (BR) are plant steroid hormones that regulate plant growth and development and modulate their reaction against many environmental stresses including temperature stress, but their role in modifying the properties of the biological membrane is poorly known. In this paper, we characterise the molecular dynamics of chloroplast membranes that had been isolated from wild-type and a BR-deficient barley mutant that had been acclimated to low and high temperatures in order to enrich the knowledge about the role of BR as regulators of the dynamics of the photosynthetic membranes. The molecular dynamics of the membranes was investigated using electron paramagnetic resonance (EPR) spectroscopy in both a hydrophilic and hydrophobic area of the membranes. The content of BR was determined, and other important membrane components that affect their molecular dynamics such as chlorophylls, carotenoids and fatty acids in these membranes were also determined. The chloroplast membranes of the BR-mutant had a higher degree of rigidification than the membranes of the wild type. In the hydrophilic area, the most visible differences were observed in plants that had been grown at 20 °C, whereas in the hydrophobic core, they were visible at both 20 and 5 °C. There were no differences in the molecular dynamics of the studied membranes in the chloroplast membranes that had been isolated from plants that had been grown at 27 °C. The role of BR in regulating the molecular dynamics of the photosynthetic membranes will be discussed against the background of an analysis of the photosynthetic pigments and fatty acid composition in the chloroplasts.
Assuntos
Brassinosteroides/metabolismo , Cloroplastos/metabolismo , Hordeum/fisiologia , Aclimatação , Cloroplastos/genética , Resposta ao Choque Frio , Resposta ao Choque Térmico , Hordeum/genética , Simulação de Dinâmica Molecular , Mutação , FotossínteseRESUMO
The integral parts of the cell membranes are the functional proteins, which are crucial for cell life. Among them, proton-pumping ATPase and aquaporins appear to be of particular importance. There is some knowledge about the effect of the temperature during plant growth, including stress-inducing temperatures, on the accumulation of the membrane proteins: plasma membrane H+-ATPase and aquaporins, but not much is known about the effect of the phytohormones (i.e. brassinosteroids (BR)) on control of accumulation of these proteins. The aim of our study was to answer the question of how a BR deficit and disturbances in the BR perception/signalling affect the accumulation of plasma membrane H+-ATPase (PM H+-ATPase), the aquaporin HvPIP1 transcript and protein in barley growing at 20⯰C and during its acclimation at 5⯰C and 27⯰C. For the studies, the BR-deficient mutant 522DK (derived from the wild-type Delisa), the BR-deficient mutant BW084 and the BR-signalling mutant BW312 and their wild-type Bowman were used. Generally, temperature of growth was significant factor influencing on the level of the accumulation of the H+-ATPase and HvPIP1 transcript and the PM H+-ATPase and HvPIP1 protein in barley leaves. The level of the accumulation of the HvPIP1 transcript decreased at 5⯰C (compared to 20⯰C), but was higher at 27⯰C than at 20⯰C in the analyzed cultivars. In both cultivars the protein HvPIP1 was accumulated in the highest amounts at 27⯰C. On the other hand, the barley mutants with a BR deficiency or with BR signalling disturbances were characterised by an altered accumulation level of PM H+-ATPase, the aquaporin HvPIP1 transcript and protein (compared to the wild types), which may suggest the involvement of brassinosteroids in regulating PM H+-ATPase and aquaporin HvPIP1 at the transcriptional and translational levels.
Assuntos
Adenosina Trifosfatases/genética , Aquaporinas/genética , Hordeum/fisiologia , Proteínas de Plantas/genética , Aclimatação , Adenosina Trifosfatases/metabolismo , Aquaporinas/metabolismo , Brassinosteroides/metabolismo , Temperatura Baixa , Hordeum/genética , Temperatura Alta , Mutação , Proteínas de Plantas/metabolismoRESUMO
A literature-based, manually-curated database of PCR primers for the detection of antibiotic resistance genes in various environments was constructed (LCPDb-ARG; lcpdb.ddg.biol.uw.edu.pl and lcpdb.ddlemb.com). Currently, this database is comprised of 607 PCR primer pairs designed for the amplification of various genes conferring resistance to antibiotics representing 10 classes of antimicrobial agents. Three parameters were assigned for each primer pair: specificity, efficacy and taxonomic efficacy. These parameters were evaluated using a novel bioinformatic tool, UniPriVal, developed for this study. UniPriVal was used to validate each primer pair against various databases, including the Bacterial Antimicrobial Resistance Reference Gene Database (BARRGDB) and those of the National Center of Biotechnology Information (NCBI). Primer pairs specific for each antibiotic resistance gene were ranked based on their model success metric value. To validate the utility and correctness of the information collected in the LCPDb-ARG, selected primer pairs were tested in bioinformatic and experimental PCR surveys. To our knowledge, this is the first database designed to facilitate PCR monitoring of the occurrence and diversity of antibiotic resistance genes in environmental and clinical samples. The internal validation system of this user-friendly application enables the quantified ranking of PCR primer pairs, which assists selection of the best primers for each application.
Assuntos
Antibacterianos , Biologia Computacional , Primers do DNA , Resistência Microbiana a Medicamentos , Reação em Cadeia da PolimeraseRESUMO
The purpose of experiments was to describe the alterations of content of steroid regulators (brassinosteroids, progesterone) during cold hardening of winter wheat. Further we studied physiological and biochemical changes induced by these steroids in cold hardened winter wheat together with estimation of plant frost tolerance. The endogenous brassinosteroid content was elevated in winter wheat during cold hardening while level of progesterone was lowered. A higher content of brassinosteroids (but not progesterone) was connected to better frost tolerance of winter wheat cultivars. Plant supplementation with brassinosteroid (24-epibrassinolide) and progesterone before cold hardening reduced frost damage. Tests with the inhibitors of the biosynthesis of brassinosteroids and progesterone suggested that these steroids are one of players in regulating the antioxidant system in winter wheat during cold hardening. Their role in regulating the expression of Rubisco or the Rubisco activase gene was less clear. Steroid regulators did not affect the content of the stress hormone ABA. Model studies of the membranes, made on a Langmuir bath, showed an increase in the value of the parameter describing differences in membrane compressibility (resulting from stronger interactions among the molecules in the monolayers). This suggests that 24-epibrassinolide and progesterone enter into the lipid layer and - in a similar way to sterols - stabilise the interaction among lipids. It may be significant step for better frost tolerance. The use of steroid regulators (especially brassinosteroids) as agrochemicals improving frost tolerance of winter cereals will be discussed.
Assuntos
Triticum/metabolismo , Temperatura Baixa , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Progesterona/metabolismo , Ribulose-Bifosfato Carboxilase/metabolismo , Triticum/genéticaRESUMO
This study focused on the idea that the toxic effect of zearalenone (ZEA) and the protective actions of the brassinosteroid - 24-epibrassinolide (EBR) as well as selenium are dependent on its accumulation in chloroplasts to a high degree. These organelles were isolated from the leaves of oxidative stress-sensitive and stress-tolerant wheat cultivars that had been grown from grains that had been incubated in a solution of ZEA (30⯵M), Na2SeO4 (Se, 10⯵M), EBR (0.1⯵M) or in a mixture of ZEA with Se or EBR. Ultra-high performance liquid chromatography techniques indicated that ZEA was adsorbed in higher amounts in the chloroplasts in the sensitive rather than tolerant cultivar. Although the brassinosteroids and Se were also accumulated in the chloroplasts, higher levels were only found in the tolerant cultivar. The application of EBR increased the homocastasterone content, especially in the chloroplasts of the tolerant plant and after the addition of ZEA. The presence of both protectants caused a decrease in the ZEA content in studied organelles and resulted in diminishing of the oxidative stress (i.e. changes in the activity of the antioxidative enzymes). Moreover, a recovery of photosystem II and decrease in the negative impact of ZEN on Hsp90 transcript accumulation was observed in plants.