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Silica nanoparticles (SiNPs) and brassinolide (BR) have been used as nano-fertilizer and growth regulator, respectively to enhance crop tolerance to abiotic stress. However, it is unclear whether a combination of the two (BR + SiNPs) is more beneficial than single application of BR or SiNPs to improve the growth of deficit-irrigated sugar beets. In this study, a two-year (2022-2023) field experiment was conducted to investigate the effects of foliar spraying of water (CK), SiNPs, BR, and BR + SiNPs on the antioxidant defense, photosynthetic capacity, dry matter accumulation, nutrient uptake, and yield of sugar beets under full irrigation (100% of crop evapotranspiration (ETc), W1) and deficit irrigation (60% ETc, W2). The results showed that compared with the application of BR or SiNPs, the application of BR + SiNPs could enhance the antioxidant defense, osmoregulation, and photosynthesis of the full-irrigated and deficit-irrigated sugar beet leaves, and ultimately improved the water status, growth, and yield of sugar beet plants. There was no significant difference in the net revenue (NR) between BR + SiNPs treatment and CK under W1 conditions. However, the NR of the BR + SiNPs treatment increased by 27.0% (p < 0.05) compared with that of CK under W2 conditions, and there was no significant difference in NR between BR + SiNPs and SiNPs treatments. A comprehensive evaluation using entropy weight combined with technique for order preference by similarity to ideal solution method found that under deficit irrigation condition, spraying SiNPs could improve the growth of sugar beet, increase the TY, NR, and water use efficiency, and reduce costs compared with spraying BR + SiNPs. Therefore, foliar spraying of SiNP on deficit-irrigated sugar beets can be used to improve sugar beet growth and reduce the potential economic losses caused by deficit irrigation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

There has been a growing concern over soil cadmium (Cd) pollution, underscoring the importance of finding effective remediation strategies. Willow trees have emerged as promising candidates for phytoremediation of Cd-contaminated soils. Nevertheless, the specific potential of a novel willow genotype, NJU513, in remediating Cd-polluted soil remains unexplored. Hence, the primary objectives of this study were twofold: firstly, to ascertain the suitability of the willow genotype NJU513 for remediating Cd-contaminated soil; and secondly, to elevate its remediation efficciency with the application of epibrassinolide (Brs). In the pot-culture experiment without Brs, its leaf and stem Cd concentrations were 203 mg kg -1 and 65.1 mg kg -1 , with a bioaccumulation factor (BCF) of 20.8 and 6.68, respectively. In the pot-culture experiment with Brs, the corresponding Cd concentrations were 226 mg kg -1 and 59.2 mg kg -1 , with a BCF of 23.1 and 6.06, respectively. In addition, the extracted Cd contents were higher in the Brs treatments (1.11-1.37 mg plant -1 ) than in the no-Brs treatments (0.78-0.96 mg plant -1 ) because Brs increased the plant biomass and leaf BCF. The mechanism underlying the Cd accumulation of NJU513 leaves with and without Brs was revealed by a transcriptome analysis. The expression levels of genes related to metal ion binding, channel activity, and transporters in leaves were up-regulated, which contributed to the high Cd accumulation and stress tolerance. Analyses of soil metabolites and bacteria in the presence and absence of Brs spraying on willow leaves indicated that soil organic compounds with carboxyl and amino groups may induce Cd activation and passivation, respectively. This study provides valuable insights for developing woody plant varieties that can be used for remediating Cd-contaminated soil., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

The introduction of arsenic, a hazardous metalloid, into the soil system due to heavy industrialization has negatively affected agricultural productivity, resulting in limited crop yields. A recent breakthrough in stress-responsive hormones, specifically brassinosteroids, has extensively covered the role of antioxidant enzyme defense systems in heavy metal stress mitigation. Considering the antioxidant properties and metal complex formation abilities of polyphenols, our study focuses on examining their role in arsenate toxicity amelioration by 24-epibrassinolide. We demonstrate enhanced growth parameters of sodium arsenate-stressed seedlings upon application of 24-epibrassinolide, with increased root and shoot polyphenol levels analyzed by high-performance liquid chromatography. Specifically, the concentration of catechin, sinapic acid, 4-hydroxy benzoic acid, protocatechuic acid, 4-coumaric acid, and myricetin were elevated, indicating induction of phenylpropanoid signaling pathway. Further, we also report a decrease in the generation of superoxide anions and hydrogen peroxide validated the antioxidant effects of these metabolites through the nitrobluetetrazolium and diaminobenzidine staining method. In addition, evaluation of transcript level of genes encoding for specific enzymes of the phenylpropanoid pathway in shoot and root showed a significant upregulation in mRNA expression of phenylalanine ammonia-lyase-1, cinnamate-4-hydroxylase, and caffeic acid o-methyltransferase-1 upon exogenous application of 24-epibrassinolide in arsenate stressed Oryza sativa., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

Salt stress is a critical limiting factor for fruit yield and quality of apples. Brassinosteroids (BRs) play an important role in response to abiotic stresses. In the present study, application of 2,4- Epicastasterone on seedlings of Malus 'M9T337' and Malus domestica 'Gala3' alleviated the physiological effects, such as growth inhibition and leaf yellowing, induced by salt stress. Further analysis revealed that treatment with NaCl induced expression of genes involved in BR biosynthesis in 'M9T337' and 'Gala3'. Among which, the expression of BR biosynthetic gene MdBR6OX2 showed a three-fold upregulation upon salt treatment, suggesting its potential role in response to salt stress in apple. MdBR6OX2, belonging to the CYP450 family, contains a signal peptide region and a P450 domain. Expression patterns analysis showed that the expression of MdBR6OX2 can be significantly induced by different abiotic stresses. Overexpressing MdBR6OX2 enhanced the tolerance of apple callis to salt stress, and the contents of endogenous BR-related compounds, such as Typhastero (TY), Castasterone (CS) and Brassinolide (BL) were significantly increased in transgenic calli compared with that of wild-type. Extopic expression of MdBR6OX2 enhanced tolerance to salt stress in Arabidopsis. Genes associated with salt stress were significantly up-regulated, and the contents of BR-related compounds were significantly elevated under salt stress. Our data revealed that BR-biosynthetic gene MdBR6OX2 positively regulates salt stress tolerance in both apple calli and Arabidopsis., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Masson SAS.)

Brassinosteroids (BR) play crucial roles in plant development and abiotic stresses in plants. Exogenous application of BR can significantly enhance cold tolerance in rice. However, the regulatory relationship between cold tolerance and the BR signaling pathway in rice remains largely unknown. Here, we characterized functions of the BR receptor OsBRI1 in response to cold tolerance in rice using its loss-of-function mutant (d61-1). Our results showed that mutant d61-1 was less tolerant to cold stress than wild-type (WT). Besides, d61-1 had lower levels than WT for some physiological parameters, including catalase activity (CAT), superoxide dismutase activity (SOD), peroxidase activity (POD), peroxidase activity (PRO), soluble protein, and soluble sugar content, while malondialdehyde content (MDA) and relative electrical conductivity (REC) levels in d61-1 were higher than those in WT plants. These results indicated that the loss of OsBRI1 function resulted in decreased cold tolerance in rice. In addition, we performed RNA sequencing (RNA-seq) of WT and d61-1 mutant under cold stress. Numerous common and unique differentially expressed genes (DEGs) with up- and down-regulation were observed in WT and d61-1 mutant. Some DEGs were expressed to various degrees, even opposite, between CK1 vs. T1 (WT) and CK2 vs. T2 (d61-1). Among these specific DEGs, some typical genes are involved in plant tolerance to cold stress. Through weighted correlation network analysis (WGCNA), 50 hub genes were screened in the turquoise and blue module. Many genes were involved in cold stress and plant hormone, such as Os01g0279800 (BRI1-associated receptor kinase 1 precursor), Os10g0513200 (Dwarf and tiller-enhancing 1, DTE1), Os02g0706400 (MYB-related transcription factor, OsRL3), etc. Differential expression levels of some genes were verified in WT and d61-1 under cold stress using qRT-PCR. These valuable findings and gene resources will be critical for understanding the regulatory relationships between cold stress tolerance and the BR signaling pathways in rice., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

The bulbil expansion of P. ternata is a key period for its yield formation, and the process of bulbil expansion is often subjected to short-term heavy precipitation. It is not clear whether the short-term waterlogging can affect bulbil expansion. Brassinolide (BR) is widely believed to enhance plant tolerance to abiotic stress. The study investigated the effects of normal water (C), waterlogging (W), waterlogging + BR (W + B), waterlogging + propiconazole (W + P) on P. ternata at the bulbil expansion period in order to assess P. ternata's ability to cope with waterlogging during the bulbil expansion stage and the regulation effects of BR on the process. The biomass of P. ternata was significantly increased after waterlogging. W treatment significantly reduced the H 2 O 2 and MDA contents, the rate of O 2 ⋅ - production and the activities of antioxidant enzymes compared with the C group. AsA and GSH contents were significantly reduced by W treatment. However, the ratios of AsA/DHA and GSH/GSSG were slightly affected by W treatment. The rate of O 2 ∙ - production and H 2 O 2 content in W + B group were significantly lower than those in W group. The POD, APX, and GR activities, and GSH content in W + B group were evidently increased compared with the W group. Soluble sugar and active ingredients contents were significantly increased after waterlogging, and the enhancement was reinforced by BR. In conclusion, waterlogging reduced oxidative stress in P. ternata under the experimental conditions. BR treatment under waterlogging had a positive effect on P. ternata by enhancing antioxidant capacity and promoting the accumulation of soluble sugars and active ingredients., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Masson SAS. All rights reserved.)

Gibberellins (GAs) play a crucial role in regulating secondary growth in angiosperms, but their effects on the secondary growth of gymnosperms are rarely reported. In this study, we administered exogenous GA 3 to two-year-old P. massoniana seedlings, and examined its effects on anatomical structure, physiological and biochemical changes, and gene expression in stems. The results showed that exogenous GA 3 could enhance xylem development in P. massoniana by promoting cell division. The content of endogenous hormone (including auxins, brassinosteroids, and gibberellins) were changed and the genes related to phytohormone biosynthesis and signaling pathway, such as GID1, DELLA, TIR1, ARF, SAUR, CPD, BR6ox1, and CYCD3, were differentially expressed under GA 3 treatment. Furthermore, GA 3 and BR (brassinosteroid) might act synergistically in promoting secondary growth in P. massoniana. Additionally, lignin content was significantly increased after GA 3 treatment accompanied by the express of lignin biosynthesis related genes. PmCAD (TRINITY&#95;DN142116&#95;c0&#95;g1), a crucial gene involved in the lignin biosynthesis, was cloned and overexpressed in Nicotiana benthamiana, significantly promoting the xylem development and enhancing stem lignification. It was regarded as a key candidate gene for improving stem growth of P. massoniana. The findings of this study have demonstrated the impact of GA 3 treatment on secondary growth of stems in P. massoniana, providing a foundation for understanding the molecular regulatory mechanism of stem secondary growth in Pinaceae seedlings and offering theoretical guidance for cultivating new germplasm with enhanced growth and yield., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Masson SAS. All rights reserved.)

Cold stress (CS) significantly limits cucumber yield. However, it remains unclear whether and how sodium nitrophenolate (CSN) regulates plant responses to cold stress. Here, H 2 O, CSN, 24-epibrassinolide (EBR), and CSN + EBR were sprayed on cucumber seedlings before or after CS, and on control plants. We found that CSN, EBR, or EBR + CSN pre-treatment improved seedling growth under normal conditions (control condition) and cold tolerance under CS conditions. EBR pre-treatment promoted the expression of approximately half of the genes involved in BR synthesis and signaling and CsICE-CsCBF-CsCOR under CS. However, CSN pre-treatment promoted almost all the expression of BR synthesis and signaling genes, and CsICE-CsCBF-CsCOR genes, which showed the highest expression in early CS, remarkably improving the cold tolerance of cucumber. Interestingly, EBR and CSN had a superimposed effect on the expression of BR synthesis and signaling and CsICE-CsCBF-CsCOR genes, which rapidly increased their expression under normal temperature. Spraying EBR after CS accelerated seedling recovery, whereas CSN had the opposite effect. However, spraying CSN combined with EBR accelerated the recovery of CS-injured seedlings and was better than spraying EBR alone. Although CS-injured seedlings were negatively influenced by CSN, pre-treatment with CSN accelerated seedling growth and increased cold tolerance, suggesting that the effect of CSN was related to whether the seedlings were damaged by CS. In conclusion, we firstly found that CSN enhanced cold tolerance by activating BR signaling, contributing to the gene expression of ICE-CBF-COR and that CSN + EBR contributed to cold tolerance and CS-injured seedling recovery in cucumber., Competing Interests: Declaration of competing interest The authors have no conflicts of interest to declare., (Copyright © 2023 Elsevier Masson SAS. All rights reserved.)

The molecular processes behind Parkinson's disease (PD) remain under debate although mitochondrial oxidative stress generation has been proposed as a fundamental contributor. In this context, different brassinosteroids have shown neuroprotective action hampering oxidative stress. This study determined the effects of 24-Epibrassinolide (24-EPI) against 6-hydroxydopamine- (6-OHDA-) induced toxicity using the zebrafish embryonic model. Embryos were exposed to 250 μM 6-OHDA or co-exposed to 24-EPI (0.01, 0.1, and 1 μM) for 3 days, starting at 48 h post-fertilization (hpf). During the experimental period, developmental parameters were assessed. At 120 hpf, larvae were tested for behavioural phenotypes with different biochemical biomarkers and tyrosine hydroxylase- (TH-) reactive neurons being also assessed. Exposure to 6-OHDA induced a decrease in body length while no other morphological phenotypes were noticed. A significant decrease in TH-neurons immunofluorescence, a decreased locomotion (speed and distance moved), and an increased absolute angle were found in 6-OHDA-exposed embryos. These outcomes were rescuable by the co-exposure with 24-EPI. Surprisingly, the direct effects of 6-OHDA on reactive oxygen species (ROS) were not observed in the present study supporting the involvement of other molecular pathways in the 6-OHDA-induced effects during embryonic development. Overall, the results obtained confirm PD-like symptoms induced by 6-OHDA during embryonic development which were reverted by 24-EPI. Although antioxidative signalling pathways deserve further scrutiny, the findings support the further investigation of 24-EPI neuroprotective effects., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.)

Nicotine is a highly addictive substance that can cause teratogenic impacts in the embryo through redox-dependent pathways. As antioxidants, naturally occurring chemicals can protect cells from redox imbalance. The purpose of this study was to evaluate the effectiveness of 24-epibrassinolide (24-EPI), a natural brassinosteroid with well-known antioxidant properties, in protecting zebrafish embryos against nicotine's teratogenic effects. For 96 h, embryos (2 h post-fertilization - hpf) were exposed to 100 μM nicotine, co-exposed with 24-EPI (0.01, 0.1, and 1 μM), and 24-EPI alone (1 μM). Lethal and sublethal developmental characteristics were evaluated during exposure. Biochemical tests were performed at the conclusion of the exposure, and distinct behavioural paradigms were analysed 24 h later. Nicotine exposure resulted in a higher proportion of larvae with deformities, which were decreased following co-exposure to 24-EPI. Nicotine exposure also caused an increase in oxidative stress as observed by the increased activity of superoxide dismutase and catalase accompanied by an increase in the malondialdehyde levels. Besides, metabolic changes were noticed as observed by the increased lactate dehydrogenase activity that were hypothesised to be associated to nicotine-induced hypoxia which may be responsible for the increased oxidative damage. In addition, locomotor deficits were observed as well as a decrease in the acetylcholinesterase activity denoting nicotine-induced cognitive dysfunction. However, co-exposure to 24-EPI alleviated behavioural deficits and improved nicotine-induced emotional states. Overall, and although further studies are required to clarify these effects, 24-EPI showed promising ameliorative properties against the teratogenic effects induced by nicotine., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Brassinosteroid (BR) has been indicated to induce the production of hydrogen peroxide (H 2 O 2 ) in plants in response to various environmental stimuli. However, it remains largely unknown how BR induces H 2 O 2 production. In this study, we found that BR treatment significantly raised the kinase activity of maize (Zea mays L.) brassinosteroid-signaling kinase 1 (ZmBSK1) using the immunoprecipitation kinase assay. ZmBSK1 could modulate the gene expressions and activities of nicotinamide adenine dinucleotide phosphate (NADPH) oxidases (EC 1.6.3.1) to modulate BR-induced H 2 O 2 production. BR could enhance the interaction between ZmBSK1 and maize calcium/calmodulin-dependent protein kinase (ZmCCaMK), a previously identified substrate of ZmBSK1. The BR-induced phosphorylation and kinase activity of ZmCCaMK are dependent on ZmBSK1. Moreover, we showed that ZmBSK1 regulated the NADPH oxidase gene expression and activity via directly phosphorylating ZmCCaMK. Genetic analysis suggested that ZmBSK1-ZmCCaMK module strengthened plant tolerance to oxidative stress induced by exogenous application of H 2 O 2 through improving the activities of antioxidant defense enzyme and alleviating the malondialdehyde (MDA) accumulation and electrolyte leakage rate. In conclusion, these findings provide the new insights of ZmBSK1 functioning in BR-induced H 2 O 2 production and the theoretical supports for breeding stress-tolerant crops., (Copyright © 2022 Elsevier Masson SAS. All rights reserved.)

Drought stress is one of the major abiotic stresses that limit grape growth and yield. Brassinosteroids (BRs) are a class of phytohormones essential for plant growth, development, and adaptation to environmental stress. This study aimed to reveal the physiological and biochemical mechanisms of exogenous BRs in alleviating the drought stress in grapevines. Two-year-old grape seedlings (Vitis vinifera L.) were sprayed with 24-epibrassinolide (EBR), a synthetic analog of BRs, and then subjected to drought treatment. The results showed that exogenous EBR significantly mitigated the reduction of photosynthetic pigment contents and photosystem II efficiency and decreased the damage to chloroplasts when grape seedlings were subjected to drought stress. Drought stress resulted in the accumulation of reactive oxidative species (ROS) and an increase in lipid peroxidation. A reduction in oxidative damage was observed in EBR-pretreated plants, which was probably due to the elevated antioxidant system. Exogenous EBR improved the activities of superoxide dismutase (14%), catalase (18%), peroxidase (17%), and ascorbate peroxidase (9%), and promoted the accumulation of ascorbic acid (10%) and glutathione (7%) under drought stress. EBR pretreatment also promoted autophagic activity, which contributed to the degradation of damaged chloroplasts. Moreover, EBR pretreatment increased the concentrations of abscisic acid, jasmonic acid, auxin, and gibberellic acid. Taken together, exogenous EBR could ameliorate the deleterious effects of drought stress by up-regulating photosynthetic capacity, antioxidant system, autophagic activity, and hormone concentrations., (Copyright © 2022. Published by Elsevier Masson SAS.)

Previous embryonic fish data have shown caffeine to induce potential teratogenic and long-term neurodevelopmental outcomes through oxidative stress-mediated apoptosis. In this context, antioxidants may have the potential to counteract the caffeine-induced effects. Therefore, the present study aimed to investigate the potential protective role of 24-epibrassinolide (24-EPI), a natural brassinosteroid with proven antioxidant properties, against caffeine-induced teratogenic effects during early zebrafish development. Embryos (~2 h post-fertilization - hpf) were exposed to 0.5 mM caffeine, co-exposed to 24-EPI (0.01, 0.1 and 1 μM) and to 24-EPI alone (1 μM) for 96 h. During exposure, lethal and sublethal developmental parameters were evaluated. At the end of the exposure, biochemical evaluations were made, and 24 h after, different behavioural paradigms were assessed. An increased number of animals showing oedema and malformations were observed after caffeine exposure, while these were reduced after co-exposure to 24-EPI concentration, namely the tail curvature. The results showed oxidative stress and related parameters similar among treatments. Yet, caffeine exposure resulted in locomotor deficits (decreased speed and distance) and disrupted anxiety-like and avoidance responses. The co-exposure to caffeine and to the highest 24-EPI concentrations resulted in less pronounced behavioural deficits. Overall, there was an absence of effects in the embryo/larvae exposed solely to 24-EPI, while caffeine caused developmental and neurotoxic effects. Although further studies are needed, the results showed promising protective effects of the highest 24-EPI concentration tested against the toxicity induced by caffeine in zebrafish., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Involvement of nitrate reductase (NR) and nitric oxide synthase (NOS)-like enzyme in 24-epibrassinolide (EB)-triggered nitric oxide (NO) synthesis to improve iron deficiency (ID) tolerance in strawberry plants was studied. EB was sprayed to strawberry plants every two days for two weeks. Then, the EB-treated plants were pre-treated with inhibitors of NR, tungstate, or NOS, L-NAME for 3 h. During the first three weeks, Fe was supplied as 100 μM EDTA-Fe or FeSO 4 to Fe-sufficient or Fe-deficient plants, respectively. Thereafter, plants were subjected for further three weeks to control (100 μM EDTA-Fe) and Fe deficiency (ID; without Fe). ID reduced biomass, chlorophyll, and chlorophyll fluorescence, while increased oxidative stress parameters, ascorbate (AsA), glutathione (GSH), endogenous NO, and the activities of NR, NOS, and antioxidant enzymes. Pre-treatments with EB and EB + SNP improved ID tolerance of strawberry by improving leaf Fe 2+ , plant growth, and antioxidant enzyme activities, and causing a further elevation in AsA, GSH, NO, NR and NOS. L-NAME application reversed NOS activity, but it did not eliminate NO, however, tungstate application reversed both NR activity and NO synthesis in plants exposed to ID + EB, suggesting that NR is the main contributor of EB-induced NO synthesis to improve ID tolerance in strawberry plants., Competing Interests: Declaration of competing interest The authors declare no conflict of interest among them., (Copyright © 2020 Elsevier Masson SAS. All rights reserved.)

Plants and crops are widely suffered by shade stress in the natural communities or in the agricultural fields. The three main phytohormones auxin, gibberellins (GAs) and brassinosteroids (BRs) were found essential in shade avoidance in Arabidopsis. However, their relationship have been seldom reported in plant shade avoidance control. Here, we report our investigation of the crosstalk of auxin, GAs and BRs in shade-induced hypocotyl elongation of soybean. Exogenous feeding of indol-3-acetic acid (IAA), GA3 or 24-epibrassinolide (EBL) distinctly promoted hypocotyl elongation in the white light, while the potent biosynthesis inhibitors of GA3, IAA, BRs severely diminished shade-induced hypocotyl elongation. Synergistic treatment of their biosynthesis inhibitors showed that GA3 fully, while EBL slightly, restored the hypocotyl elongation that was efficiently repressed by IAA biosynthesis inhibitor, GA3 and IAA dramatically suppressed the hypocotyl growth inhibition by BR biosynthesis inhibitor in the shade, whereas both IAA and EBL feeding cannot suppress the elongation inhibition by GA biosynthesis inhibitor. Further analyses revealed that shade remarkably upregulated expression of key genes of IAA, GA and BR biosynthesis in the soybean hypocotyls, and GA biosynthesis genes were effectively blocked by IAA, GA and BR biosynthesis inhibitors in the shade. Taken together, these results suggest that GAs modulate shade-induced hypocotyl elongation downstream of mutual promotion of auxin and BRs in soybean., Competing Interests: Declaration of competing interest The authors declare that they have no conflicts of interest., (Copyright © 2020 Elsevier Masson SAS. All rights reserved.)

Salinity stress reduces growth and yield productivity of most crop plants. Potentiality of kinetin (Kn) and epi-brassinolide (EBL), either individually or combinedly in preventing the salinity (100 mM NaCl) stress mediated oxidative damage and photosynthetic inhibition was studied in Solanum lycopersicum. Combined application of Kn and EBL imparted much prominent impact on the growth, photosynthesis and metabolism of antioxidants, osmolytes and secondary metabolites. Synthesis of chlorophylls and carotenoids increased and the photosynthetic parameters like stomatal conductance, intercellular CO 2 concentration and net photosynthesis were significantly improved due to application of Kn and EBL. Photosystem II functioning (Fv/Fm), photochemical quenching and electron transport rate (ETR) improved significantly in Kn and EBL treated plants imparting significant decline in salinity induced non-photochemical quenching. Exogenous Kn and EBL effectively prevented the oxidative damage by significantly declining the generation of hydrogen peroxide and superoxide under saline and non-saline conditions as reflected in lowered lipid peroxidation and electrolyte leakage. Reduced oxidative damage in Kn and EBL treated plants was accompanied down-regulation of protease and lipoxygenase concomitant with up-regulation of the antioxidant system and the accumulation of compatible osmolytes. Treatment of Kn and EBL proved effective in enhancing the contents of redox homeostasis, ascorbic acid and reduced glutathione, and the secondary metabolites assisting the enzymatic antioxidant system in combating the salinity stress efficiently. Results suggest that combined application of Kn and EBL regulate growth and photosynthesis in tomato more effectively than their individual application through a probable regulatory crosstalk mechanism., Competing Interests: Declaration of competing interest The authors declare that no conflict of interest exists., (Copyright © 2019. Published by Elsevier Masson SAS.)

Brassinosteroids (BRs) play a crucial role in improving plant resistance to various environmental stresses. In this study, we aimed to explore the potential role of BRs in protecting plants from antimony (Sb) toxicity. In the in vitro agar-plate culture experiments, the level changes of BR in wide-type plants and BR biosynthesis mutant dwrf4-1 significantly affected the corresponding response of Arabidopsis to Sb stress. Increasing the BR content significantly enhanced Sb-induced root growth inhibition and lowering the BR level appeared to reduce the plant sensitivity to Sb stress. Foliar application of eBL, however, significantly decreased the Sb accumulation and peroxidation of membrane lipids, increased the contents of chlorophyll and proline, and further boosted and strengthened the antioxidant enzymes activities. These experiments demonstrated that BRs played an important role in regulating heavy metal stress responses in plants and exogenous foliar spray of eBL was an important method for alleviating toxicity of Sb., (Copyright © 2019 Elsevier Masson SAS. All rights reserved.)

Salinity is a common environmental challenge limiting worldwide agricultural crop yield. Plants employ epigenetic regulatory strategies, such as DNA methylation which relatively allows rapid adaptation to new conditions in response to environmental stresses. Brassinosteroids (BRs) are a novel group of phytohormones recognized as transcription and translation regulators which are able to mitigate the impact of environmental stresses on the plants. In the current investigation, the influence of salinity and 24-epibrassinolide (24-epiBL) was investigated on the extent and pattern of cytosine DNA methylation using methylation-sensitive amplified polymorphisms (MSAP) technique in flax. Upon NaCl (150 mM) exposure, total methylation of CCGG sequences was decreased in comparison to control plants, while 24-epiBL (10 -8  M) induced total methylation under salinity stress. Sequencing and analysis of six randomly selected MSAP fragments detected genes involved in various biological and molecular processes such as vitamine B1 biosynthesis, protein targeting and localization, post-translational modification and gene regulation. In conclusion, 24-epiBL seed priming could play critical role in regulation of cellular and biological processes in response to salt stress by epigenetic modification and induction of methylation., (Copyright © 2019 Elsevier Masson SAS. All rights reserved.)

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 μM), Na 2 SeO 4 (Se, 10 μM), EBR (0.1 μM) 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., (Copyright © 2019 Elsevier Masson SAS. All rights reserved.)

Soil salinity is one of the major abiotic stress factors that hampers plant growth and productivity by limiting photosynthesis and other related metabolic processes. In this study we investigated whether treatment with proline and/or 24-epibrassinolide (EBL) to two contrasting cultivars of Brassica juncea (L.) Czern and Coss viz. Varuna and RH-30 could counteract with the adverse effects of salinity on photosynthesis and seed yield. Plants were treated with proline and/or 24-epibrassinolide (EBL) at 28 and 29d-stages of growth. Salt stress reduced plant growth, photosynthetic attributes, efficiency of PSII (Fv/Fm), leaf water potential and finally seed yield, at harvest but improved the activity of antioxidant enzymes in both the cultivars in a concentration dependent manner. Exogenous application of EBL with proline completely neutralised the adverse effects of salt at 78 mM or 117 mM stress levels whereas the treatment partially neutralised the impact of highest salt concentration of 156 mM, through the upregulation of the antioxidant system., (Copyright © 2019 Elsevier Masson SAS. All rights reserved.)

Drought stress is one of most dramatic abiotic stresses, reduces crop yield significantly. Application of hormones proved as an effective drought stress ameliorating approach. 24-Epibrassinolide (EBL), an active by-product from brassinolide biosynthesis increases drought stress tolerance in plants significantly. EBL application enhances plant growth and development under drought stress by acting as signalling compound in different physiological processes. This article discussed potential role of 24-epibrassinolide application and drought tolerance in plants. Briefly, EBL sustains or improves plant growth and yield by enhancing carbon assimilation rate, maintaining a balance between ROS and antioxidants and also plays important role in solute accumulation and water relations. Furthermore, we also compared different EBL application methods and concluded that seed priming and foliar application are more productive as compared with root application method. In conclusion, EBL is very impressive phyto-hormone, which can ameliorate drought stress induced detrimental effects in plants., (Crown Copyright © 2018. Published by Elsevier Masson SAS. All rights reserved.)

Drought stress is a serious threat to agriculture and the environment. Brassinosteroids (BRs) increase tolerance to drought stress of plant. Autophagy plays important roles in plant responses to drought stress; however, there are few reports on autophagy in peach (Prunus persica). In total, 23 putative autophagy-related genes (ATGs) in peach were identified using ATGs from the Arabidopsis thaliana genome as query in BLASTx algorithm-based searches. Under drought stress, the photosynthetic abilities of peach leaves decreased, while antioxidant enzyme activities, autophagy and ATG expression increased. A correlation analysis showed that antioxidant enzyme activities are inversely correlated to the expression levels of the PpATGs. During drought, the PpATG8s and some PpATG18s had the strongest responses. To investigate enhanced drought-stress tolerance, peach was treated with water, 100 nM 24-epibrassinolide (EBR), 1 μM EBR, 10 μM EBR and 1 μM voriconazole. Exogenous EBR at 1 μM decreased the malondialdehyde (MDA) content under drought stress when compared with water-, 1 μM voriconazole-, 100 nM EBR- and 10 μM EBR-treated peach leaf. The 1-μM EBR application increased superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), ascorbate peroxidase (APX) and glutathione peroxidase (GR) activities during drought stress. In addition, the expression levels of PpATGs were inhibited by EBR. Thus, the 1-μM EBR treatment alleviated drought-stress damage to peach leaves, decreased PpATG expression levels and reduced the number of autophagosomes., (Copyright © 2018. Published by Elsevier Masson SAS.)

Copper (Cu) stress is the most common abiotic stress experienced in vineyards owing to the copper-based fungicides application. Plant hormones, including 24-Epibrassinolide (EBR), may alleviate the adverse impacts of heavy metal stress on plants. We investigated the effects of EBR pretreatment on root morphological parameters, active oxygen metabolism, osmolytes contents, antioxidant enzyme activity, endogenous phytohormone contents, and ascorbate-glutathione (AsA-GSH) cycle activity of one-year-old grape (Vitis vinifera L.) cuttings under Cu stress. Pretreatment with EBR significantly enhanced root morphological parameters (total root length, root surface area, root diameter, root volume, and tip number), increased soluble protein and proline contents, and significantly decreased the contents of H 2 O 2 , O 2 ⋅- , and malondialdehyde (MDA) in roots and leaves. EBR pretreatment increased the activities of superoxide dismutase (SOD), catalase (CAT), peroxidase oxidase (POD), and the contents of the endogenous phytohormones abscisic acid, jasmonic acid, and salicylic acid in the leaves. In addition, EBR regulated the balance of the AsA-GSH cycle by increasing the activities of monodehydroascorbate reductase (MDHAR), glutathione peroxidase (GR), ascorbate peroxidase (APX), and dehydroascorbate reductase (DHAR), and the contents of the antioxidant ascorbate (AsA) and dehydroascorbic acid (DHA), but the contents of glutathione (GSH) and oxidized glutathione (GSSG) decreased. Among the treatments tested, pretreatment with 0.10 mg/L EBR showed the optimal performance for alleviation of Cu toxicity. The results show that exogenous brassinosteroids reduce oxidative damage and improve the tolerance of Cu stress of grapevine cuttings., (Copyright © 2018 Elsevier Masson SAS. All rights reserved.)

BRs are polyhydroxylated sterol derivatives, classified as phytohormones. Plants of Brassica juncea var. Varuna were grown in pots and an aqueous solution (10 -8  M) of two brassinosteroid isomers 28-homobrassinolide (HBL) and 24-epibrassinolide (EBL) of same concentration (10 -8  M) was applied to their leaves. The treatment up-regulated the photosynthetic machinery directly by enhancing water splitting activity, photochemical quenching, non-photochemical quenching, maximum PSII efficiency, actual PSII efficiency, electron transport rate, stomatal movement, stomatal conductance, internal CO 2 concentration, transpiration rate, net photosynthetic rate and carbohydrate synthesis. Moreover, the level of biochemical enzymes (carbonic anhydrase and nitrate reductase), reactive oxygen species (superoxide and hydrogen peroxide) generation, antioxidant enzyme activity and mineral status (C, N, Mg, P, S, K), which indirectly influence the rate of photosynthesis, also improved in the treated plants. Out of the two BR analogues tested, EBL excelled in its effects over HBL., (Copyright © 2018 Elsevier Masson SAS. All rights reserved.)

Brassinosteroids play an important role in the plant growth and development as well as in the adaptation of plants to environmental stresses. Studies have shown the effect of 24-epibrassinolide (EBL) in the range of concentrations 0.0001-10 μM on the green unicellular alga Acutodesmus obliquus (Chlorophyceae) during 7 days of cultivation. EBL is an effective stimulator of algal growth as it causes an increase in the number of cells and the contents of selected metabolites such as proteins, monosaccharides, and photosynthetic pigments (chlorophylls, carotenes, and xanthophylls). Furthermore, EBL inhibits the formation of reactive oxygen species such as hydrogen peroxide and oxidative damage as evidenced by a decrease of the lipid peroxidation. The positive effect of EBL resulting from the cellular oxidative state can be alleviated by antioxidants such as ascorbate peroxidase, catalase, superoxide dismutase, and ascorbate., (Copyright © 2018 Elsevier Masson SAS. All rights reserved.)

Brassinolide (BL) and castasterone (CS) are the representative members of brassinosteroid class of plant steroid hormone having plant growth promoting activity. In this study, eleven CS analogs bearing a variety of side chains were synthesized to determine the effect of the side chain structures on the BL-like activity. The plant hormonal activity was evaluated in a dwarf rice lamina inclination assay, and the potency was determined as the reciprocal logarithm of the 50% effective dose (ED 50 ) from each dose-response curve. The reciprocal logarithm of ED 50 (pED 50 ) was decreased dramatically upon deletion of the C-28 methyl group of CS. The introduction of oxygen-containing groups such as hydroxy, methoxy, and ethoxycarbonyl was also unfavorable to the activity. The pED 50 was influenced by the geometry of carbon-carbon double bond between C-24 and C-25 (cis and trans), but the introduction of a fluorine atom at the C-25 position of the double bond did not significantly change the activity. The binding free energy (ΔG) was calculated for all ligand-receptor binding interactions using molecular dynamics, resulting that ΔG is linearly correlated with the pED 50 ., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Presence of the toxic metalloid, "arsenic (As)" is ubiquitous in the environment especially in the soil and water. Its excess availability in the soil retards growth and metabolism of plants via (a) slowing down the cell division/elongation, (b) overproduction of reactive oxygen species (ROS), (c) modulation of antioxidant enzymes, and (d) alteration of DNA profile/genomic template stability (GTS). In the current study, diphenylene iodonium (DPI), 24-epibrassinolide (EBL) and proline (Pro) were used to analyze their roles in eliminating the adverse effects of As. Glycine max L. (variety JS 335) seeds were subjected to As (75 μM, Sodium arsenite was used as source of As), and in combination with DPI (10 μM), EBL (0.5 μM) or Pro (10 mM), for five consecutive days, and effects of these treatment combinations were analyzed on germination percentage, biomass, membrane stability, GTS and expressions of defensive genes. In addition, the levels of As, ROS, malondialdehyde, DNA content, oxidation, fragmentation, polymorphism, DNase activity, endogenous Pro and pyrroline-5-carboxylate synthetase activity were evaluated. The results indicated that the treatments of DPI, EBL or Pro are capable to alleviate detrimental effects of As, gauged from above variables, but with different magnitudes. Apropos As-stress mitigation, Pro was found to be the most effective under the confines of the study protocol. This study certainly provides new ideas for intensifying studies to unravel elusive central mechanism of amelioration involving use of DPI, EBL or Pro in plants with confirmed As-toxicity., (Copyright © 2016 Elsevier Masson SAS. All rights reserved.)

Brassinosteroids (BRs) are steroid plant hormones that have been shown to be involved in the response to salt stress in cross-talk with other plant growth regulators such as polyamines (PAs). In addition, BRs are involved in the regulation of the nodulation in the rhizobium-legume symbiosis through the alteration of the PAs content in leaves. In this work, we have studied the effect of exogenous 24-epibrassinolide (EBL) in the response to salinity of nitrogen fixation in the symbiosis Medicago truncatula-Sinorhizobium meliloti. Foliar spraying of EBL restored the growth of plants subjected to salt stress and provoked an increment of the nitrogenase activity. In general, PAs levels in leaves and nodules decreased by the salt and EBL treatments, however, the co-treatment with NaCl and EBL augmented the foliar spermine (Spm) concentration. This increment of the Spm levels was followed by a reduction of the membrane oxidative damage and a diminution of the proline accumulation. The effect of BRs on the symbiotic interaction was evaluated by the addition of 0.01, 0.1 and 0.5 μM EBL to the growing solution, which provoked a reduction of the nodule number and an increment of the PAs levels in shoot. In conclusion, foliar treatment with EBL had a protective effect against salt stress in the M. truncatula-S. meliloti symbiosis mediated by an increment of the Spm levels. Treatment of roots with EBL incremented PAs levels in shoot and reduced the nodule number which suggests a cross-talk between PAs and BRs in the nodule suppression and the protection against salt stress., (Copyright © 2016 Elsevier Masson SAS. All rights reserved.)

In this study, we performed a comparative analysis of the physiological and biochemical parameters of wheat cultivars with contrasting drought resistance, drought-resistant Omskaya 35 (O-35) and less drought-resistant Salavat Yulaev (SYu), during 7-day germination under drought stress simulated by 5% mannitol. In addition, we evaluated the effectiveness of pre-sowing seed treatment with 0.4 μM 24-epibrassinolide (EBR) used to increase the resistance of plants of both cultivars to drought stress. It was revealed that mannitol has caused significant changes in the hormonal balance of the plants of both cultivars, associated with abscisic acid (ABA) accumulation and decrease in the contents of indoleacetic acid (IAA) and cytokinins (CKs). It should be noted that more dramatic changes in the content of phytohormones were characteristic for seedlings of SYu cultivar, which was reflected in a stronger growth inhibition of these plants. Pretreatment with EBR mitigated the negative effect of drought on the hormonal status and growth of seedlings during their germination. Furthermore, we found that drought caused accumulation of dehydrin (DHN) proteins, especially of low molecular weight DHNs, whose abundance was 2.5 times greater in O-35 cultivar than in SYu plants. EBR-pretreated plants of both cultivars were characterized by the additional accumulation of DHNs, indicating their involvement in the development of the EBR-induced wheat drought resistance. The use of fluridone allowed us to demonstrate ABA-dependent and ABA-independent pathways of regulation of low molecular mass dehydrins accumulation by EBR in wheat plants of both cultivars under drought conditions., (Copyright © 2016 Elsevier Masson SAS. All rights reserved.)

In the present study, we aimed to investigate the effects of exogenous 24-epibrassinolide (EBR) on growth, photosynthetic characteristics, chlorophyll fluorescence imaging, and nitrogen metabolism of tomato leaves under low temperature and weak light conditions. The results showed that foliar application of EBR significantly alleviated the inhibition of plant growth, and increased the fresh and dry weights of tomato plants under a combined low temperature and weak light stress. Moreover, EBR also increased the net photosynthetic rate (Pn), light saturation point (LSP), maximal quantum yield of PSII photochemistry (Fv/Fm), actual photochemical efficiency of PSII (ФPSII), and photochemical quenching coefficient (qP), but decreased the intercellular CO2 concentration (Ci), light compensation point (LCP) and apparent quantum efficiency (AQE) under low temperature and weak light conditions. In addition, application of EBR to tomato leaves significantly enhanced the activities of nitrate reductase (NR), glutamate dehydrogenase (GDH), glutamine synthetase (GS), and glutamate synthase (GOGAT), but decreased the ammonium content and nitrite reductase (NiR) activity. We observed that EBR remarkably increased the contents of aspartic acid, threonine, serine, glycine, and phenylalanine, while decreasing the accumulation of cysteine, methionine, arginine, and proline under a combined low temperature and light stress. These results suggest that EBR could alleviate the combined stress-induced harmful effects on photosynthesis and nitrogen metabolism, thus leading to improved plant growth., (Copyright © 2016. Published by Elsevier Masson SAS.)

Sugar unloading in grape berries is a crucial step in the long-distance transport of carbohydrates from grapevine leaves to berries. Brassinosteroids (BRs) mediate many physiological processes in plants including carbohydrate metabolism. Here, 'Cabernet Sauvignon' (Vitis vinifera L.) grape berries cultivated in clay loam fields were treated with an exogenous BR (24-epibrassinolide; EBR), a BR synthesis inhibitor (brassinazole; Brz), Brz + EBR (sprayed with EBR 24 h after a Brz treatment), and deionized water (control) at the onset of véraison. The EBR treatment sharply increased the soluble sugars content in the berries, but decreased it in the skins. The EBR and Brz + EBR treatments significantly promoted the activities of both invertases (acidic and neutral) and sucrose synthase (sucrolytic) at various stages of ripening. The mRNA levels of genes encoding sucrose metabolic invertase (VvcwINV), and monosaccharide (VvHT3, 4, 5 and 6) and disaccharide (VvSUC12 and 27) transporters were increased by the EBR and/or Brz + EBR treatments. Generally, the effects of the Brz treatment on the measured targets contrasted with the effects of the EBR treatments. The EBR and Brz treatments inhibited the biosynthesis of the endogenous BRs 6-deoxocastastarone and castasterone. Both EBR and Brz + EBR treatments increased the brassinolide contents, down-regulated the expression of genes encoding BRs biosynthetic enzymes BRASSINOSTEROID-6-OXIDASE and DWARF1, (VvBR6OX1 and VvDWF1) and induced BR receptor gene BRASSINOSTEROID INSENSITIVE 1 (VvBRI1) expression in deseeded berries. Together, these results show that BRs are involved in controlling sugar unloading in grape berries during véraison., (Copyright © 2015 Elsevier Masson SAS. All rights reserved.)

In the present work the effects of exogenous 24-epibrassinolide (EBR) on functional and structural characteristics of the thylakoid membranes under non-stress conditions were evaluated 48 h after spraying of pea plants with different concentrations of EBR (0.01, 0.1 and 1.0 mg.L(-1)). The results show that the application of 0.1 mg.L(-1) EBR has the most pronounced effect on the studied characteristics of the photosynthetic membranes. The observed changes in 540 nm light scattering and in the calorimetric transitions suggest alterations in the structural organization of the thylakoid membranes after EBR treatment, which in turn influence the kinetics of oxygen evolution, accelerate the electron transport rate, increase the effective quantum yield of photosystem II and the photochemical quenching. The EBR-induced changes in the photosynthetic membranes are most probably involved in the stress tolerance of plants., (Copyright © 2014 Elsevier Masson SAS. All rights reserved.)

Interaction between brassinosteroids (BRs) (brassinolide, BL; 24-epibrassinolide, 24-epiBL; 28-homobrassinolide, 28-homoBL; castasterone, CS; 24-epicastasterone, 24-epiCS; 28-homocastasterone, 28-homoCS) and adenine- (trans-zeatin, tZ; kinetin, Kin) as well as phenylurea-type (1,3-diphenylurea, DPU) cytokinins (CKs) in the regulation of cell number, phytohormone level and the content of chlorophyll, monosaccharide and protein in unicellular green alga Chlorella vulgaris (Trebouxiophyceae) were examined. Chlorella vulgaris exhibited sensitivity to CKs in the following order of their stimulating properties: 10 nM tZ > 100 nM Kin >1 μM DPU. Exogenously applied BRs possessed the highest biological activity in algal cells at concentration of 10 nM. Among the BRs, BL was characterized by the highest activity, while 28-homoCS - by the lowest. The considerable increase in the level of all endogenous BRs by 27-46% was observed in C. vulgaris culture treated with exogenous 10 nM tZ. It can be speculated that CKs may stimulate BR activity in C. vulgaris by inducing the accumulation of endogenous BRs. CKs interacted synergistically with BRs increasing the number of cells and endogenous accumulation of proteins, chlorophylls and monosaccharides in C. vulgaris. The highest stimulation of algal growth and the contents of analyzed biochemical parameters were observed for BL applied in combination with tZ, whereas the lowest in the culture treated with both 28-homoCS and DPU. However, regardless of the applied mixture of BRs with CKs, the considerable increase in cell number and the metabolite accumulation was found above the level obtained in cultures treated with any single phytohormone in unicellular green alga C. vulgaris., (Copyright © 2014 Elsevier Masson SAS. All rights reserved.)

The relationships between brassinosteroids (BRs) (brassinolide, BL; 24-epiBL; 28-homoBL; castasterone, CS; 24-epiCS; 28-homoCS) and auxins (indole-3-acetic acid, IAA; indole-3-butyric acid, IBA; indole-3-propionic acid, IPA) in the regulation of cell number, phytohormone level and metabolism in green alga Chlorella vulgaris were investigated. Exogenously applied auxins had the highest biological activity in algal cells at 50 μM. Among the auxins, IAA was characterized by the highest activity, while IBA - by the lowest. BRs at 0.01 μM were characterized by the highest biological activity in relation to auxin-treated and untreated cultures of C. vulgaris. The application of 50 μM IAA stimulated the level of all detected endogenous BRs in C. vulgaris cells. The stimulatory effect of BRs in green algae was arranged in the following order: BL > 24-epiBL > 28-homoBL > CS > 24-epiCS > 28-homoCS. Auxins cooperated synergistically with BRs stimulating algal cell proliferation and endogenous accumulation of proteins, chlorophylls and monosaccharides in C. vulgaris. The highest stimulation of algal growth and the contents of analyzed biochemical parameters were observed for the mixture of BL with IAA, whereas the lowest in the culture treated with both 28-homoCS and IBA. However, regardless of the applied mixture of BRs with auxins, the considerable increase in cell number and the metabolite accumulation was found above the level obtained in cultures treated with any single phytohormone. Obtained results confirm that both groups of plant hormones cooperate synergistically in the control of growth and metabolism of unicellular green alga C. vulgaris., (Copyright © 2013 Elsevier Masson SAS. All rights reserved.)

Plant steroidal hormones, brassinosteroids, play a pivotal role in variety of plant developmental processes and adaptation to various environmental stresses. The present work investigates the response of various stress markers upon exogenous application of 24-epibrassinolide (EBL) on Pusa Basmati-1, a commercially important rice variety, under salt stress conditions. Rice seeds after treatment with different concentrations of NaCl alone or in combination with different concentrations of 24-epibrassinolide (EBL) were analysed for various growth parameters, protein, proline and malondialdehyde content (MDA) and antioxidant enzymes activities. The seedlings exposed to NaCl exhibited a significant decline in growth parameters and changes in the levels of antioxidant enzymes, however, treatment with EBL showed an improvement in growth, levels of protein and proline content and antioxidant enzymes activity. The enhanced levels of MDA content during salt stress in rice seedlings was decreased with EBL treatment. Further, the treatment with EBL increased the expression of various oxidative stress marker genes, although to different levels. Expression of various brassinosteroids (OsBRI1, OsDWF4) and salt (SalT) responsive genes, revealed the down regulation of OsDWF4 with application of EBL and upregulation of SalT in presence of salt stress thereby confirming the efficacy of the treatments. Interestingly, a significant down regulation of SalT gene was observed on application of EBL along with salt compared to salt treatment alone. On the other hand, the application of EBL alone and in combination with salt has resulted in upregulation of OsBRI1., (Copyright © 2013 Elsevier Masson SAS. All rights reserved.)

A novel chemical tool compound that is an antagonist of brassinolide (BL, 1)-induced rice lamina joint inclination was developed. Although 2-O-, 3-O-, 22-O-, or 23-O-methylation of BL causes a critical decrease in biological activity,(5) a crystal structure of the extracellular leucine-rich repeat (LRR) domain of BRASSINOSTEROID-INSENSITIVE I (BRI1) bound to BL(3,4) indicates that the loss of activity of the O-methylated BL may result from not only the low affinity to BRI1, but also from blocking the interaction with another BR signaling factor, a partner protein of BRI1 (e.g., BRI1-ASSOCIATED KINASE 1, BAK1). On the basis of this hypothesis we synthesized the BL 2,3-acetonide 2, the 22,23-acetonide 3, and the 2,3:22,23-diacetonide 4 to assess the possibility of 2-O- and 3-O- or/and 22-O- and 23-O-alkylated BL as an antagonist in BR signaling evoked by exogenously applied BL. The 2,3-acetonide 2 more strongly inhibited the lamina inclination caused by BL relative to the 22,23-acetonide 3, whereas the diacetonide 4 had no effect most likely due to its increased hydrophobicity. This suggested that the 2,3-hydroxyl groups of BL play a more significant role in the interaction with a BRI1 partner protein rather than BRI1 itself in rice lamina joint inclination. Taken together it was demonstrated that BL, the most potent agonist of BRI1, is transformed into an antagonist by functionalization of the 2,3-dihydroxyl groups as the acetonide. This finding opens the door to the potential development of a chemical tool that modulates protein-protein interactions in the BR signaling pathway to dissect the BR-dependent processes., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Ca(NO(3))(2) accumulation is a major factor that limits greenhouse production in China. The present investigation was carried out to study the effect of 24-epibrassinolide (EBL) on nitrogen metabolism (including contents of NO(3)(-), NH(4)(+) and amino acids and related enzymes activities) in cucumber seedlings (Cucumis sativus L. cv. Jinyou No. 4) under 80 mM Ca(NO(3))(2) stress. This study found that exogenous EBL significantly reduced the accumulation of NO(3)(-) and NH(4)(+) by Ca(NO(3))(2), and enhanced the inactivated enzymes activities involved in the nitrogen metabolism. In addition, EBL alleviated the inhibition of photosynthesis nitrogen-use efficiency by Ca(NO(3))(2). Increased total amino acids by EBL under stress increased the precursor of proteins biosynthesis, thus promoting the biosynthesis nitrogen containing compounds. The presence of Ca(NO(3))(2) increased polyamines level, which might result from the increased content of free putrescine that is harmful to plant growth. However, exogenous EBL induced a further increase in total polyamines. The increase is likely caused by the elevated contents of conjugated and bound forms of polyamines. In summary, exogenously EBL compensated for the damage/losses by Ca(NO(3))(2) stress to some extent through the regulation of nitrogen metabolism and metabolites., (Copyright © 2012. Published by Elsevier Masson SAS.)

The objective of this study was to explore the response of 24-epibrassinolide to improve the biological yield of Ni-tolerant and Ni-sensitive varieties of Vigna radiata and also to test the propositions that 24-epibrassinolide induced up-regulation of antioxidant system protects the efficiency of V. radiata, grown under Ni-stress. Surface sterilized seeds of var. T-44 (Ni-tolerant) and PDM-139 (Ni-sensitive) were soaked in DDW (control), 10(-10), 10(-8), or 10(-6) M of 24-epibrassinolide for 8 h (shotgun approach). These treated seeds were then inoculated with specific Rhizobium grown in sandy loam soil supplemented with different levels of Ni 0, 50, 100, or 150 mg Ni kg(-1) of soil and were allowed to grow for 45-days. At this stage of growth, plants were sampled to assess the various growths and nodule related traits as well as selected biochemical characteristics. The remaining plants were allowed to grow to maturity to study the yield characteristics. The results indicated that plant-fresh and dry mass, number of nodules, their fresh and dry mass, leghemoglobin content, nitrogen and carbohydrate content in the nodules, leaf chlorophyll content, activities of nitrate reductase and carbonic anhydrase decreased proportionately with the increasing concentrations of soil nickel. However, the application of 24-epibrassinolide as shotgun approach (pre-sowing seed soaking) to the nickel-stressed or non-stressed plants improved growth, nodulation and enhanced the activity of various antioxidant enzymes (viz. catalase, peroxidase and superoxide dismutase) and also the content of proline. The up-regulation of antioxidant enzymes as well as proline (osmolyte) triggered by 24-epibrassinolide could have conferred tolerance to the Ni-stressed plants resulting in improved growth, nodulation and yield attributes., (Copyright © 2012 Elsevier Masson SAS. All rights reserved.)

Fast and stable 2-fold accumulation of cytokinins (CKs) was detected initially in roots and then in shoots of 4-day-old wheat (Triticum aestivum L.) seedlings in the course of their treatment with 0.4μM 24-epibrassinolide (EBR). Elevated cytokinin level has been maintained only in the presence of EBR, while the hormone removal has led to return of cytokinin concentration to the control level initially in the roots and then in the shoots. EBR-induced accumulation of cytokinins was accompanied by inhibition of both cytokinin oxidase (CKX) (cytokinin oxidase/dehydrogenase, EC 1.5.99.12) activity and expression of the gene coding for this enzyme, and on the contrary the decline in CKs level resulted in increase in these characteristics up to the control level in roots and then in shoots. Sharp accumulation of cytokinin O-glucosides has been discovered in response to EBR-treatment suggesting fast EBR-induced activation of production of cytokinins, which excessive amounts were transferred into the storage forms. The obtained data provide evidence for the involvement of EBR in regulation of cytokinin level in wheat seedlings., (Copyright © 2012 Elsevier Masson SAS. All rights reserved.)

Brassinosteroids (BRs) are plant steroid hormones and, when applied exogenously, they induce physiological responses, including tolerance to heat shock (HS). How endogenous BR content and altered perception of BRs influence thermal tolerance is poorly understood. BR-induced thermotolerance in tomato seedlings with altered BR homeostasis was examined by assessing the survival, ion leakage and lipid peroxidation of seedlings from a BR-deficient mutant (extreme dwarf d(x)), a partially BR-insensitive mutant curl3(-abs) allele (curl3 altered brassinolide sensitivity) and a line overexpressing the Dwarf, BR-biosynthesis gene (35SD). We confirmed that treatment with 1 μM of epi-brassinolide (EBL) induces thermotolerance of wild type seedlings following a HS regime at 45 °C. The curl3(-abs) seedlings had the highest basal tolerance to heat, whereas the EBL-induced thermal tolerance of d(x) seedlings was greatest and responded to lower EBL concentrations. The d(x) and 35SD seedlings had similar thermal tolerance; however, they showed increased signs of oxidative stress. EBL reduced the induction of lipid peroxidation of seedlings after recovery from heat. Highest oxidative stress and peroxidase (POX) activity (EC 1.11.1.7) was in BR-deficient d(x) mutant seedlings. EBL was able of inducing POX activity but not other antioxidant enzymes; however, effects of HS on POX activity of seedlings were absent or less marked. Taking together, results indicate that thermal tolerance is independent of endogenous BR content, but HS-mediated oxidative stress depends on BR levels., (Copyright © 2011 Elsevier Masson SAS. All rights reserved.)

To identify nuclear proteins involved in the brassinosteroid (BR) signaling pathway, a targeted proteomic approach was applied to Arabidopsis thaliana suspension-cultured T87 cells. Cell growth was promoted by 0.1 μM brassinolide (BL) and inhibited by 5 μM brassinazole (Brz). Analysis of BR-regulated proteins in nuclear-enriched fractions was carried out using two-dimensional polyacrylamide gel electrophoresis with a special fluorescent dye. Proteins of interest were identified by correlating normalized spot volume of proteins on the gels with cellular BR level (Brz-treated cells, extremely low level of BRs; control cells, normal level of BRs; BL-treated cells, high level of BRs). A number of BR-responsive proteins were detected and some of these proteins were identified by nano-liquid chromatography-tandem mass spectrometry after enzymatic digestion. Fluctuations in eight identified nuclear proteins in BL-treated cells were investigated in the first 12 h of treatment. Three nuclear BR-responsive proteins, Nucleosome Assembly Protein (NAP) 1;1, Band 7 Family Protein, and Vernalization Independence 3, significantly decreased during this time. Meanwhile, NAP1;2, S-Adenosylmethionine Synthetase 2, and 60S Ribosomal Protein L14 increased markedly over time. Since some of these proteins are reportedly related to chromosome remodeling, cell growth induced by BL may involve chromatin remodeling. Interestingly, NAP1;2 was found to be post-translationally modified in response to cellular BR levels. Our study of quantitative protein changes in the nucleus provides valuable insight into BR-induced cellular and physiological responses., (Copyright © 2011 Elsevier Masson SAS. All rights reserved.)