Showing papers on "Brassinosteroid published in 2005"
TL;DR: It is shown that BZR1 is a transcriptional repressor that has a previously unknown DNA binding domain and binds directly to the promoters of feedback-regulated BR biosynthetic genes.
Abstract: Brassinosteroid (BR) homeostasis and signaling are crucial for normal growth and development of plants. BR signaling through cell-surface receptor kinases and intracellular components leads to dephosphorylation and accumulation of the nuclear protein BZR1. How BR signaling regulates gene expression, however, remains unknown. Here we show that BZR1 is a transcriptional repressor that has a previously unknown DNA binding domain and binds directly to the promoters of feedback-regulated BR biosynthetic genes. Microarray analyses identified additional potential targets of BZR1 and illustrated, together with physiological studies, that BZR1 coordinates BR homeostasis and signaling by playing dual roles in regulating BR biosynthesis and downstream growth responses.
706 citations
TL;DR: It is demonstrated that brassinosteroids bind directly to the 94 amino acids comprising ID-LRR22 in the extracellular domain of BRI1, and define a new binding domain for steroid hormones.
Abstract: Both animals and plants use steroids as signalling molecules during growth and development. Animal steroids are principally recognized by members of the nuclear receptor superfamily of transcription factors. In plants, BRI1, a leucine-rich repeat (LRR) receptor kinase localized to the plasma membrane, is a critical component of a receptor complex for brassinosteroids. Here, we present the first evidence for direct binding of active brassinosteroids to BRI1 using a biotin-tagged photoaffinity castasterone (BPCS), a biosynthetic precursor of brassinolide (the most active of the brassinosteroids). Binding studies using BPCS, (3)H-labelled brassinolide and recombinant BRI1 fragments show that the minimal binding domain for brassinosteroids consists of a 70-amino acid island domain (ID) located between LRR21 and LRR22 in the extracellular domain of BRI1, together with the carboxy-terminal flanking LRR (ID-LRR22). Our results demonstrate that brassinosteroids bind directly to the 94 amino acids comprising ID-LRR22 in the extracellular domain of BRI1, and define a new binding domain for steroid hormones.
598 citations
TL;DR: Multiple lines of evidence together suggest that the D11/CYP724B1 gene plays a role in BR synthesis and may be involved in the supply of 6-DeoxoTY and TY in the BR biosynthesis network in rice.
Abstract: We have characterized a rice (Oryza sativa) dwarf mutant, dwarf11 (d11), that bears seeds of reduced length. To understand the mechanism by which seed length is regulated, the D11 gene was isolated by a map-based cloning method. The gene was found to encode a novel cytochrome P450 (CYP724B1), which showed homology to enzymes involved in brassinosteroid (BR) biosynthesis. The dwarf phenotype of d11 mutants was restored by the application of the brassinolide (BL). Compared with wild-type plants, the aberrant D11 mRNA accumulated at higher levels in d11 mutants and was dramatically reduced by treatment with BL, implying that the gene is feedback-regulated by BL. Precise determination of the defective step(s) in BR synthesis in d11 mutants proved intractable because of tissue specificity and the complex control of BR accumulation in plants. However, 6-deoxotyphasterol (6-DeoxoTY) and typhasterol (TY), but not any upstream intermediates before these compounds, effectively restored BR response in d11 mutants in a lamina joint bending assay. Multiple lines of evidence together suggest that the D11/CYP724B1 gene plays a role in BR synthesis and may be involved in the supply of 6-DeoxoTY and TY in the BR biosynthesis network in rice.
462 citations
TL;DR: Quantitative analysis suggested that in brd2, the 24-methylene BR biosynthesis pathway is activated and the uncommon BR, dolichosterone (DS), is produced, indicating that DS is a bioactive BR in rice.
Abstract: We have identified a rice (Oryza sativa) brassinosteroid (BR)-deficient mutant, BR-deficient dwarf2 (brd2). The brd2 locus contains a single base deletion in the coding region of Dim/dwf1, a homolog of Arabidopsis thaliana DIMINUTO/DWARF1 (DIM/DWF1). Introduction of the wild-type Dim/dwf1 gene into brd2 restored the normal phenotype. Overproduction and repression of Dim/dwf1 resulted in contrasting phenotypes, with repressors mimicking the brd2 phenotype and overproducers having large stature with increased numbers of flowers and seeds. Although brd2 contains low levels of common 6-oxo-type BRs, the severity of the brd2 phenotype is much milder than brd1 mutants and most similar to d2 and d11, which show a semidwarf phenotype at the young seedling stage. Quantitative analysis suggested that in brd2, the 24-methylene BR biosynthesis pathway is activated and the uncommon BR, dolichosterone (DS), is produced. DS enhances the rice lamina joint bending angle, rescues the brd1 dwarf phenotype, and inhibits root elongation, indicating that DS is a bioactive BR in rice. Based on these observations, we discuss an alternative BR biosynthetic pathway that produces DS when Dim/dwf1 is defective.
265 citations
TL;DR: The hypothesis that 23-O-glucosylation of BL is a function of UGT73C5 in planta, and that glucosylations regulates BR activity is supported, is supported.
Abstract: Steroid hormones are essential for development, and the precise control of their homeostasis is a prerequisite for normal growth. UDP-glycosyltransferases (UGTs) are considered to play an important regulatory role in the activity of steroids in mammals and insects. This study provides an indication that a UGT accepting plant steroids as substrates functions in brassinosteroid (BR) homeostasis. The UGT73C5 of Arabidopsis thaliana catalyses 23-O-glucosylation of the BRs brassinolide (BL) and castasterone. Transgenic plants overexpressing UGT73C5 displayed BR-deficient phenotypes and contained reduced amounts of BRs. The phenotype, which was already apparent in seedlings, could be rescued by application of BR. In feeding experiments with BL, wild-type seedlings converted BL to the 23-O-glucoside; in the transgenic lines silenced in UGT73C5 expression, no 23-O-glucoside was detected, implying that this UGT is the only enzyme that catalyzes BL-23-O-glucosylation in seedlings. Plant lines in which UGT73C5 expression was altered also displayed hypocotyl phenotypes previously described for seedlings in which BR inactivation by hydroxylation was changed. These data support the hypothesis that 23-O-glucosylation of BL is a function of UGT73C5 in planta, and that glucosylation regulates BR activity.
225 citations
TL;DR: The results imply that BR homeostasis is maintained through feedback expressions of multiple genes, each of which is involved not only in BR-specific biosynthesis and inactivation, but also in sterol biosynthesis.
Abstract: Homeostasis of brassinosteroids (BRs) is essential for normal growth and development in higher plants. We examined responsiveness of 11 BR metabolic gene expressions to the decrease or increase of endogenous BR contents in Arabidopsis (Arabidopsis thaliana) to expand our knowledge of molecular mechanisms underlying BR homeostasis. Five BR-specific biosynthesis genes (DET2, DWF4, CPD, BR6ox1, and ROT3) and two sterol biosynthesis genes (FK and DWF5) were up-regulated in BR-depleted wild-type plants grown under brassinazole, a BR biosynthesis inhibitor. On the other hand, in BR-excessive wild-type plants that were fed with brassinolide, four BR-specific synthesis genes (DWF4, CPD, BR6ox1, and ROT3) and a sterol synthesis gene (DWF7) were down-regulated and a BR inactivation gene (BAS1) was up-regulated. However, their response to fluctuation of BR levels was highly reduced (DWF4) or nullified (the other eight genes) in a bri1 mutant. Taken together, our results imply that BR homeostasis is maintained through feedback expressions of multiple genes, each of which is involved not only in BR-specific biosynthesis and inactivation, but also in sterol biosynthesis. Our results also indicate that their feedback expressions are under the control of a BRI1-mediated signaling pathway. Moreover, a weak response in the mutant suggests that DWF4 alone is likely to be regulated in other way(s) in addition to BRI1 mediation.
209 citations
TL;DR: Results suggest that elicitation of growth promotion by PGPR in Arabidopsis is associated with several different signal transduction pathways and that such signaling may be different for plants grown in vitro vs. in vivo.
Abstract: Plant growth-promoting rhizobacteria (PGPR) colonize plant roots and exert beneficial effects on plant health and development. We are investigating the mechanisms by which PGPR elicit plant growth promotion from the viewpoint of signal transduction pathways within plants. We report here our first study to determine if well-characterized PGPR strains, which previously demonstrated growth promotion of various other plants, also enhance plant growth in Arabidopsis thaliana. Eight different PGPR strains, including Bacillus subtilis GB03, B. amyloliquefaciens IN937a, B. pumilus SE-34, B. pumilus T4, B. pasteurii C9, Paenibacillus polymyxa E681, Pseudomonas fluorescens 89B-61, and Serratia marcescens 90-166, were evaluated for elicitation of growth promotion of wild-type and mutant Arabidopsis in vitro and in vivo. In vitro testing on MS medium indicated that all eight PGPR strains increased foliar fresh weight of Arabidopsis at distances of 2, 4, and 6 cm from the site of bacterial inoculation. Among the eight strains, IN937a and GB03 inhibited growth of Arabidopsis plants when the bacteria were inoculated 2 cm from the plants, while they significantly increased plant growth when inoculated 6 cm from the plants, suggesting that a bacterial metabolite that diffused into the agar accounted for growth promotion with this strain. In vivo, eight PGPR strains promoted foliar fresh weight under greenhouse conditions 4 weeks after sowing. To define signal transduction pathways associated with growth promotion elicited by PGPR, various plant-hormone mutants of Arabidopsis were evaluated in vitro and in vivo. Elicitation of growth promotion by PGPR strains in vitro involved signaling of brassinosteroid, IAA, salicylic acid, and gibberellins. In vivo testing indicated that ethylene signaling was involved in growth promotion. Results suggest that elicitation of growth promotion by PGPR in Arabidopsis is associated with several different signal transduction pathways and that such signaling may be different for plants grown in vitro vs. in vivo.
205 citations
TL;DR: It is postulate that castasterone is the major active brassinosteroid during vegetative growth in tomato, whereas brassinolide may play an organ-specific role in fruit development in this species.
191 citations
TL;DR: The root and nodulation phenotypes of various mutant lines of pea (Pisum sativum) that display alterations in their phytohormone levels and/or perception were characterized in this article.
Abstract: The initiation and development of legume nodules induced by compatible Rhizobium species requires a complex signal exchange involving both plant and bacterial compounds. Phytohormones have been implicated in this process, although in many cases direct evidence is lacking. Here, we characterize the root and nodulation phenotypes of various mutant lines of pea (Pisum sativum) that display alterations in their phytohormone levels and/or perception. Mutants possessing root systems deficient in gibberellins (GAs) or brassinosteroids (BRs) exhibited a reduction in nodule organogenesis. The question of whether these reductions represent direct or indirect effects of the hormone deficiency is addressed. For example, the application of GA to the roots of a GA-deficient mutant completely restored its number of nodules to that of the wild type. Grafting studies revealed that a wild-type shoot or root also restored the nodule number of a GA-deficient mutant. These findings suggest that GAs are required for nodulation. In contrast, the shoot controlled the number of nodules that formed in graft combinations of a BR-deficient mutant and its wild type. The root levels of auxin and GA were similar among these latter graft combinations. These results suggest that BRs influence a shoot mechanism that controls nodulation and that the root levels of auxin and GA are not part of this process. Interestingly, a strong correlation between nodule and lateral root numbers was observed in all lines assessed, consistent with a possible overlap in the early developmental pathways of the two organs.
170 citations
TL;DR: In this article, the authors reported the isolation of a second activation-tagged mutant with a BR-deficient phenotype caused by the overexpression of SOB7 (CYP72C1), a homolog of BAS1.
Abstract: Active brassinosteroids (BRs), such as brassinolide (BL) and castasterone (CS), are growth-promoting plant hormones An Arabidopsis cytochrome P450 monooxygenase (CYP734A1, formerly CYP72B1), encoded by the BAS1 gene, inactivates BRs and modulates photomorphogenesis BAS1 was identified as the overexpressed gene responsible for a dominant, BR-deficient mutant, bas1-D This mutant was isolated in an activation-tagged screen designed to identify redundant genes that might not be identified in classic loss-of-function screens Here we report the isolation of a second activation-tagged mutant with a BR-deficient phenotype The mutant phenotype is caused by the overexpression of SOB7 (CYP72C1), a homolog of BAS1 We generated single and double null-mutants of BAS1 and SOB7 to test the hypothesis that these two genes act redundantly to modulate photomorphogenesis BAS1 and SOB7 act redundantly with respect to light promotion of cotyledon expansion, repression of hypocotyl elongation and flowering time in addition to other phenotypes not regulated by light We also provide biochemical evidence to suggest that BAS1 and SOB7 act redundantly to reduce the level of active BRs, but have unique mechanisms Overexpression of SOB7 results in a dramatic reduction in endogenous CS levels, and although single null-mutants of BAS1 and SOB7 have the same level of CS as the wild type, the double null-mutant has twice the amount Application of BL to overexpression lines of BAS1 or SOB7 results in enhanced metabolism of BL, though only BAS1 overexpression lines confer enhanced conversion to 26-OHBL, suggesting that SOB7 and BAS1 convert BL and CS into unique products
158 citations
TL;DR: A clear connection between brassinosteroid and fiber development is established and the door for genetic analysis of cotton development is opened through direct modification of the brass inosteroid signal transduction pathway.
Abstract: ;Our current understanding of the role of phytohormones in the development of cotton fibers is derived largely from an amenable culture system in which cotton ovules, collected on the day of anthesis, are floated on liquid media Under these conditions, supplemental auxin and gibberellin were found to promote fiber initiation and elongation More recently, addition of low concentrations of the brassinosteroid brassinolide (BL) were also found to promote fiber elongation while a brassinosteroid biosynthesis inhibitor brassinazole2001 (Brz) inhibited fiber development In order to elucidate the role of brassinosteroid in cotton fiber development further, we have performed a more detailed analysis of the effects of these chemicals on cultured cotton ovules Our results confirm that exogenous BL promotes fiber elongation while treatment with Brz inhibits it Furthermore, treatment of cotton floral buds with Brz results in the complete absence of fiber differentiation, indicating that BR is required for fiber initiation as well as elongation Expression of fiber genes associated with cell elongation increased in ovules treated with BL and was suppressed by Brz treatment, establishing a correlation between brassinosteroid-regulated gene expression and fiber elongation These results establish a clear connection between brassinosteroid and fiber development and open the door for genetic analysis of cotton development through direct modification of the brassinosteroid signal transduction pathway
TL;DR: In this article, the ROTUNDIFOLIA3 (ROT3) gene, encoding the enzyme CYP90C1, which was specifically involved in the regulation of leaf length in Arabidopsis thaliana, was required for the late steps in the BR biosynthesis pathway.
Abstract: Brassinosteroids (BRs) are plant hormones that are essential for a wide range of developmental processes in plants. Many of the genes responsible for the early reactions in the biosynthesis of BRs have recently been identified. However, several genes for enzymes that catalyze late steps in the biosynthesis pathways of BRs remain to be identified, and only a few genes responsible for the reactions that produce bioactive BRs have been identified. We found that the ROTUNDIFOLIA3 (ROT3) gene, encoding the enzyme CYP90C1, which was specifically involved in the regulation of leaf length in Arabidopsis thaliana, was required for the late steps in the BR biosynthesis pathway. ROT3 appears to be required for the conversion of typhasterol to castasterone, an activation step in the BR pathway. We also analyzed the gene most closely related to ROT3, CYP90D1, and found that double mutants for ROT3 and CYP90D1 had a severe dwarf phenotype, whereas cyp90d1 single knockout mutants did not. BR profiling in these mutants revealed that CYP90D1 was also involved in BR biosynthesis pathways. ROT3 and CYP90D1 were expressed differentially in leaves of A. thaliana, and the mutants for these two genes differed in their defects in elongation of hypocotyls under light conditions. The expression of CYP90D1 was strongly induced in leaf petioles in the dark. The results of the present study provide evidence that the two cytochrome P450s, CYP90C1 and CYP90D1, play distinct roles in organ-specific environmental regulation of the biosynthesis of BRs.
TL;DR: It is shown here that tomato plants treated with 24-epibrassinolide (EBR) are more tolerant to high temperature than untreated plants, and EBR treated tomato plants had better photosynthetic efficiency.
Abstract: Brassinosteroids are naturally occurring plant growth regulators, which exhibit structural similarities to animal steroid hormones. Recent studies have indicated that besides an essential role in plant growth and development, brassinosteroids also exert anti-stress effects on plants. We show here that tomato plants treated with 24-epibrassinolide (EBR) are more tolerant to high temperature than untreated plants. An analysis of mitochondrial small heat shock proteins (MT-sHSP) in tomato leaves by western blotting revealed that the MT-sHSP did not preferentially accumulate in EBR treated plants at 25 °C. However, treatment of plants at 38 °C induced much more accumulation of MT-sHSP in EBR treated than in untreated plants. Results of this study provide the first direct evidence for EBR induced expression of MT-sHSP, which possibly induced thermotolerance in tomato plants. EBR treated tomato plants had better photosynthetic efficiency. We also observed significantly higher in vitro pollen germination, enhanced pollen tube growth and low pollen bursting in the presence of EBR at 35 °C, a temperature high enough to induce heat-stress symptoms in tomato, indicating a possible role of EBR during plant reproduction.
TL;DR: It is shown that DWF1 is a Ca2+/calmodulin-binding protein and this binding is critical for its function, and this results raise the possibility of producing size-engineered crops by altering the Ca2+, calmodulin, binding property of theirDWF1 orthologues.
Abstract: Brassinosteroids are plant steroid hormones that couple environmental factors, such as light, with plant growth and development. New work in Arabidopsis shows that to function normally, the DWARF1 protein (DWF1) required for brassinosteroid biosynthesis must bind to the Ca2+ binding protein calmodulin. The DWF1 equivalent proteins from other plants have a similar Ca2+/calmodulin-binding domain. The finding raises the possibility of genetically engineering crop plants of varying sizes to cope with in particular environmental conditions. Brassinosteroids are plant-specific steroid hormones1,2 that have an important role in coupling environmental factors, especially light, with plant growth and development3. How the endogenous brassinosteroids change in response to environmental stimuli is largely unknown. Ca2+/calmodulin has an essential role in sensing and transducing environmental stimuli4,5. Arabidopsis DWARF1 (DWF1) is responsible for an early step in brassinosteroid biosynthesis that converts 24-methylenecholesterol to campesterol6,7. Here we show that DWF1 is a Ca2+/calmodulin-binding protein and this binding is critical for its function. Molecular genetic analysis using site-directed and deletion mutants revealed that loss of calmodulin binding completely abolished the function of DWF1 in planta, whereas partial loss of calmodulin binding resulted in a partial dwarf phenotype in complementation studies. These results provide direct proof that Ca2+/calmodulin-mediated signalling has a critical role in controlling the function of DWF1. Furthermore, we observed that DWF1 orthologues from other plants have a similar Ca2+/calmodulin-binding domain, implying that Ca2+/calmodulin regulation of DWF1 and its homologues is common in plants. These results raise the possibility of producing size-engineered crops by altering the Ca2+/calmodulin-binding property of their DWF1 orthologues.
TL;DR: Montoya et al. as mentioned in this paper showed that the patterns of Dwarf expression and brassinosteroid accumulation in tomato reveal the importance of branchedosteroid synthesis during fruit development.
Abstract: Montoya, T., Nomura, T., Yokota, T., Farrar, K., Harrison, K, Jones, J. G. D., Kaneta, T., Kamiya, Y., Szekeres, M., Bishop, G. J. (2005). Patterns of Dwarf expression and brassinosteroid accumulation in tomato reveal the importance of brassinosteroid synthesis during fruit development. Plant Journal, 42, (2), 262-269. Sponsorship: BBSRC grant P13908/Human Frontier Research Program (HFSP) to GB and TY / Royal Society, STA in Japan / the Hungarian Scientific Research Fund (Grant T 42639) and travel grants from the Royal Society and the British Council
TL;DR: In this paper, a review of the physiological and molecular mechanisms underlying BR-promoted growth in different plant organs is presented, focusing on BR-sensitive genes and their interactions with other phytohormones.
Abstract: Brassinosteroids (BRs) are highly potent growth-promoting sterol derivatives. BR-deficient or BR-insensitive mutants display dwarfism. Whole plants and excised tissues have been used to analyse the mechanisms involved in BR-promoted growth. BR stimulates cell elongation and cell division, and BR has specific effects on differentiation. Underlying physiological pathways include modification of cell wall properties, effects on carbohydrate assimilation and allocation, and control of aquaporin activities. BR apparently coordinates and integrates diverse processes required for growth, partly via interactions with other phytohormones setting the frame for BR responses. Ultimately, BR-promoted growth is mediated through genomic pathways. Positive regulators of the BR response (such as BZR1 and BES1) and putative downstream components (such as EXO) are involved in the regulation of BR-responsive genes and growth promotion. BR-responsive genes have been identified in several plant species. However, causal links between physiological effects and changes of transcript patterns, for the most part, are still unresolved. This review focuses on physiology and molecular mechanisms underlying BR-promoted growth in the different plant organs. Interactions with other phytohormones are discussed.
TL;DR: In this paper, an Arabidopsis dwarf mutant, shrink 1-D (shk1-D), was isolated and the phenotype was shown to be caused by activation of the CYP72C1 gene.
Abstract: *Summary Brassinosteroids (BRs) are plant steroidal hormones that regulate plant growth and development. An Arabidopsis dwarf mutant, shrink1-D (shk1-D), was isolated and the phenotype was shown to be caused by activation of the CYP72C1 gene. CYP72C1 is a member of the cytochrome P 450 monooxygenase gene family similar to BAS1/CYP734A1 that regulates BR inactivation. shk1-D has short hypocotyls in both light and dark, and short petioles and siliques. The seeds are also shortened along the longitudinal axis indicating CYP72C1 controls cell elongation. The expression of CPD, TCH4 and BAS1 were altered in CYP72C1 overexpression transgenic lines and endogenous levels of castasterone, 6-deoxocastasterone and 6-deoxotyphasterol were also altered. Unlike BAS1/CYP734A1 the expression of CYP72C1 was not changed by application of exogenous brassinolide. We propose that CYP72C1 controls BR homeostasis by modulating the concentration of BRs.
TL;DR: The results demonstrate that the Arabidopsis CYP51A2 gene encodes a functional obtusifoliol 14α-demethylase enzyme and plays an essential role in controlling plant growth and development by a sterol-specific pathway.
Abstract: CYP51 exists in all organisms that synthesize sterols de novo. Plant CYP51 encodes an obtusifoliol 14α-demethylase involved in the postsqualene sterol biosynthetic pathway. According to the current gene annotation, the Arabidopsis (Arabidopsis thaliana) genome contains two putative CYP51 genes, CYP51A1 and CYP51A2. Our studies revealed that CYP51A1 should be considered an expressed pseudogene. To study the functional importance of the CYP51A2 gene in plant growth and development, we isolated T-DNA knockout alleles for CYP51A2. Loss-of-function mutants for CYP51A2 showed multiple defects, such as stunted hypocotyls, short roots, reduced cell elongation, and seedling lethality. In contrast to other sterol mutants, such as fk/hydra2 and hydra1, the cyp51A2 mutant has only minor defects in early embryogenesis. Measurements of endogenous sterol levels in the cyp51A2 mutant revealed that it accumulates obtusifoliol, the substrate of CYP51, and a high proportion of 14α-methyl-Δ8-sterols, at the expense of campesterol and sitosterol. The cyp51A2 mutants have defects in membrane integrity and hypocotyl elongation. The defect in hypocotyl elongation was not rescued by the exogenous application of brassinolide, although the brassinosteroid-signaling cascade is apparently not affected in the mutants. Developmental defects in the cyp51A2 mutant were completely rescued by the ectopic expression of CYP51A2. Taken together, our results demonstrate that the Arabidopsis CYP51A2 gene encodes a functional obtusifoliol 14α-demethylase enzyme and plays an essential role in controlling plant growth and development by a sterol-specific pathway.
TL;DR: A rice (Oryza sativa) dwarf mutant, dwarf and gladius leaf 1 (dgl1), which exhibits only minimal response to GA and BR, is reported, and the function of DGL1 is important for cell and organ elongation in rice, and aberrant D GL1-mediated microtubule organization causes up-regulation of gibberellin biosynthetic genes independently of gibBerell in signaling.
Abstract: Molecular genetic studies of plant dwarf mutants have indicated that gibberellin (GA) and brassinosteroid (BR) are two major factors that determine plant height; dwarf mutants that are caused by other defects are relatively rare, especially in monocot species Here, we report a rice (Oryza sativa) dwarf mutant, dwarf and gladius leaf 1 (dgl1), which exhibits only minimal response to GA and BR In addition to the dwarf phenotype, dgl1 produces leaves with abnormally rounded tip regions Positional cloning of DGL1 revealed that it encodes a 60-kD microtubule-severing katanin-like protein The protein was found to be important in cell elongation and division, based on the observed cell phenotypes GA biosynthetic genes are up-regulated in dgl1, but the expression of BR biosynthetic genes is not enhanced The enhanced expression of GA biosynthetic genes in dgl1 is not caused by inappropriate GA signaling because the expression of these genes was repressed by GA3 treatment, and degradation of the rice DELLA protein SLR1 was triggered by GA3 in this mutant Instead, aberrant microtubule organization caused by the loss of the microtubule-severing function of DGL1 may result in enhanced expression of GA biosynthetic genes in that enhanced expression was also observed in a BR-deficient mutant with aberrant microtubule organization These results suggest that the function of DGL1 is important for cell and organ elongation in rice, and aberrant DGL1-mediated microtubule organization causes up-regulation of gibberellin biosynthetic genes independently of gibberellin signaling
TL;DR: YP72C1, together with BAS1/CYP72B1, is speculated to regulate active brassinosteroid levels in plants, and expression analysis suggested that wild-type CYP 72C1 transcript levels increased after exposure to white light, although the physiological significance of such a response remains obscure.
Abstract: A pool of Arabidopsis lines transformed with the activation vector was screened for short hypocotyl mutants under dim far-red light, and three mutant lines designated chibi1-3 (chi) were isolated. Among the chi mutants, chi2 was dominant. The chi2 seedlings were short, regardless of the light conditions. The chi2 mature plants exhibited phenotypic features such as dwarfism, reduced male fertility and dark green, rounded epinastic leaves, which are characteristics of brassinosteroid-deficient mutants such as det2, cpd, and dwf4. Furthermore, the hypocotyl phenotype was restored by the addition of brassinolide to the culture medium, suggesting that brassinosteroids had been affected in this mutant. The molecular analysis of the chi2 mutant revealed that the CYP72C1 gene was overexpressed by the enhancing activity of the inserted DNA. Wild-type plants that were transformed with a vector containing a chimeric gene between the 35S promoter and the CYP72C1 genomic DNA exhibited a similar phenotype. Consistent with the morphological and physiological phenotype, the levels of active brassinosteroids were reduced in the chi2 mutant. Hence, CYP72C1, together with BAS1/CYP72B1, is speculated to regulate active brassinosteroid levels in plants. Expression analysis suggested that wild-type CYP72C1 transcript levels increased after exposure to white light, although the physiological significance of such a response remains obscure.
TL;DR: Results indicate that BRs actually negatively regulate GA20 levels in pea, and appears that the BR growth response is not mediated by changes in bioactive GA levels, thus providing further evidence thatBRs are important regulators of stem elongation.
Abstract: The objective of this study was to increase our understanding of the relationship between brassinosteroids (BRs) and gibberellins (GAs) by examining the effects of BR deficiency on the GA biosynthesis pathway in several tissue types of pea (Pisum sativum L.). It was suggested recently that, in Arabidopsis, BRs act as positive regulators of GA 20-oxidation, a key step in GA biosynthesis [Bouquin et al. (2001) Plant Physiol 127:450-458]. However, this may not be the case in pea as GA20 levels were consistently higher in all shoot tissues of BR-deficient (lk and lkb) and BR-response (lka) mutants. The application of brassinolide (BL) to lkb plants reduced GA20 levels, and metabolism studies revealed a reduced conversion of GA19 to GA20 in epi-BL-treated lkb plants. These results indicate that BRs actually negatively regulate GA20 levels in pea. Although GA20 levels are affected by BR levels, this does not result in consistent changes in the level of the bioactive GA, GA1. Therefore, even though a clear interaction exists between endogenous BR levels and the level of GA20, this interaction may not be biologically significant. In addition to the effect of BRs on GA levels, the effect of altered GA1 levels on endogenous BR levels was examined. There was no significant difference in BR levels between the GA mutants and the wild type (wt), indicating that altered GA1 levels have no effect on BR levels in pea. It appears that the BR growth response is not mediated by changes in bioactive GA levels, thus providing further evidence that BRs are important regulators of stem elongation.
TL;DR: It is found that elongated, a previously identified photomorphogenesis mutant, enhances high-light phototropism and represents a unique allele of BAK1/SERK3, a receptor kinase implicated in brassinosteroid perception.
Abstract: The phototropic response of Arabidopsis (Arabidopsis thaliana) is induced by the phototropin photoreceptors and modulated by the cryptochrome and phytochrome photoreceptors. Downstream of these photoreceptors, asymmetric lateral redistribution of auxin underlies the differential growth, which results in phototropism. Historical physiological evidence and recent analysis of hormone-induced gene expression demonstrate that auxin and brassinosteroid signaling function interdependently. Similarly, in this study we report evidence that interactions between brassinosteroids and auxin signaling modulate phototropic responsiveness. We found that elongated, a previously identified photomorphogenesis mutant, enhances high-light phototropism and represents a unique allele of BAK1/SERK3, a receptor kinase implicated in brassinosteroid perception. Altogether, our results support the hypothesis that phototropic responsiveness is modulated by inputs that influence control of auxin response factor-mediated transcription.
TL;DR: The results indicate that brassinosteroids may regulate the nodule number in soybean plants and brassinazole inhibited internodal growth in Enrei.
Abstract: Leguminous plants form nitrogen-fixing root nodules and the number of nodules is controlled by a self-regulating mechanism called autoregulation. However, signaling substances involved in nodule regulation have not been identified. In the present study, we used brassinolide, a most effective molecular species of plant hormone brassinosteroids, and brassinazole, an effective inhibitor of brassinosteroid biosynthesis to determine whether brassinolide played a role in systemic regulation of noduel formation in wild type soybean and its super-nodulating mutant. Foliar application or direct injection of brassinolide into the root base inhibited nodule formation and root development in the super-nodulating mutant (En6500), but not in the parent line (cv. Enrei). The internodes in the plants subjected to foliar application were significantly longer than those in the untreated plants. In contrast, the application of brassinazole on mature leaves or into the culture media resulted in the increase of the nodule num...
TL;DR: Findings on the effects of BRs on plant growth, BR biosynthesis and catabolism, and BR signal transduction are described.
Abstract: In animals, a large number of steroid hormones play important roles in numerous processes including reproduction and differentiation. The biologically active plant steroid brassinolide (BL) was first discovered in the pollen of western rape in 1979 (Grove et al., 1979). This finding suggested that BL is indispensable for plant growth and differentiation. To date, more than 50 BL analogs have been identified, and the group has been termed brassinosteroids (BRs) (Fujioka and Yokota, 2003). Brassinosteroids have several biological activities, such as inducing cell elongation when applied at very low concentrations. For this reason, soon after their discovery, they were suggested to be a sixth type of plant hormone; however, for years BRs were not considered true plant hormones. The turning point in BR research was the discovery of the Arabidopsis dwarf mutants det2 and cpd in 1996 (Li et al., 1996; Szekeres et al., 1996). These BR-deficient mutants were found to revert to the wild-type phenotype following BR treatment. Concurrent with the analysis of these mutants, an outline of the biosynthetic pathway of BRs was being elucidated through chemical analysis. Following the isolation of det2 and cpd, a great number of BR-deficient mutants were identified. The mutant genes were found to encode proteins that catalyze the conversion of plant steroids to BR precursors. Eventually, BRs were widely recognized as important plant hormones indispensable for growth and differentiation (Clouse and Sasse, 1998). In parallel, mutants that are insensitive to BRs were isolated (Clouse et al., 1996; Li et al., 1997) with phenotypes very similar to those of the BR-biosynthesis mutants. Investigations of these mutants revealed several mechanisms of BR perception and signal transduction (Bishop and Koncz, 2002; Clouse, 2002). This review describes findings on the effects of BRs on plant growth, BR biosynthesis and catabolism, and BR signal transduction.
TL;DR: Promoter activities of the two Arabidopsis BR C-6 oxidase genes (CYP85A1 and CYP85A2) in transgenic plants carrying promoter fusions with the GUS, GFP or LUC reporter genes were studied to gain insight into potential sites of BR synthesis.
Abstract: Brassinosteroids (BRs) are steroid hormones that are essential for plant growth and development. To gain insight into potential sites of BR synthesis, we studied promoter activities of the two Arabidopsis BR C-6 oxidase genes (CYP85A1 and CYP85A2) in transgenic plants carrying promoter fusions with the GUS, GFP or LUC reporter genes. BR-dependent feedback regulation of the GUS reporter constructs indicated that their expression corresponded to those of the native genes. Both the CYP85A1 and CYP85A2 promoters showed maximum activity during the first week following germination, particularly in the vascular tissues. Compared to CYP85A2, CYP85A1 expression was weaker and confined to the early stages of seedling development. Stronger CYP85A2 promoter activity was evident in both juvenile and adult plants. Comparison of the 5′-UTR and TATA box sequences of CYP85A1 and CYP85A2 revealed high homology, indicating a relatively recent gene duplication. We also found that transgenic Arabidopsis plants harbouring the tomato DWARF promoter-GUS fusion had similarities in the expression pattern to the Arabidopsis genes suggesting common transcriptional regulation of CYP85 genes in the two species.
TL;DR: Data demonstrate that BRS1 is a secreted and active serine carboxypeptidase, consistent with the hypothesis suggested by the previous genetic evidence that B RS1 may process a protein involved in an early event in the BRI1 signaling pathway.
TL;DR: Recent studies have begun to unravel the biochemical details of individual steps of the brassinosteroid signal transduction pathway, including ligand binding and receptor dimerization at the cell surface, signal transmission across the cell membrane, the phosphorylation of cellular targets in the cytosol, and gene regulation inside the nucleus.
TL;DR: Ulastructural studies revealed that chloroplast development was completely suppressed by chilling treatment, and a cDNA library originating from 28°C-light-grown seedling leaves was constructed for screening cold-suppressed (cos) genes.
Abstract: Etiolated leaves of 28°C-dark-grown mung bean (Vigna radiata L. cv. 2937) seedlings fail to turn green after being shifted to a light and cold environment. At the visible phenotypic level, incapability of leaf greening is the only failure event for the de-etiolation of mung bean seedlings at low temperature. Ultrastructural studies revealed that chloroplast development was completely suppressed by chilling treatment. A cDNA library originating from 28°C-light-grown seedling leaves was constructed for screening cold-suppressed (cos) genes. Thirteen full-length cDNA clones were obtained, with 12 clones encoding chloroplast proteins, which, according to their known physiological functions, were important for chloroplast development and photosynthesis. Another cos cDNA encodes CYP90A2, which is a cytochrome P450 protein involved in the biosynthesis of brassinosteroid hormones. All cos genes are light-regulated at normal temperature. The influence of chilling stress on cos expression was examined in 10°C-light- and 10°C-dark-grown etiolated seedlings, and in 10°C-light-grown green plants. The data show that cos expression in these three treatments is severely suppressed. This suppression is controlled at the transcriptional level, as demonstrated by nuclear runoff experiments, and is reversible because cos mRNAs accumulate again after the cold-treated plants have been transferred to 28°C.
TL;DR: Functional characterization of CYP51G1-Sc expressed in yeast showed that it could demethylate obtusifoliol, as well as nontypical plant sterol biosynthetic intermediates (lanosterol), in contrast with the strong substrate specificity of the previously characterized obtUSifolioiol 14α-demethylases found in other plant species.
Abstract: The sterol 14α-demethylase (CYP51) is the most widely distributed cytochrome P450 gene family being found in all biological kingdoms. It catalyzes the first step following cyclization in sterol biosynthesis, leading to the formation of precursors of steroid hormones, including brassinosteroids, in plants. Most enzymes involved in the plant sterol biosynthesis pathway have been characterized biochemically and the corresponding genes cloned. Genes coding for enzymes promoting substrate modifications before 24-methylenelophenol lead to embryonic and seed defects when mutated, while mutants downstream the 24-methylenelophenol intermediate show phenotypes characteristic of brassinosteroid mutants. By a differential display approach, we have isolated a fertilization-induced gene, encoding a sterol 14α-demethylase enzyme, named CYP51G1-Sc. Functional characterization of CYP51G1-Sc expressed in yeast (Saccharomyces cerevisiae) showed that it could demethylate obtusifoliol, as well as nontypical plant sterol biosynthetic intermediates (lanosterol), in contrast with the strong substrate specificity of the previously characterized obtusifoliol 14α-demethylases found in other plant species. CYP51G1-Sc transcripts are mostly expressed in meristems and in female reproductive tissues, where they are induced following pollination. Treatment of the plant itself with obtusifoliol induced the expression of the CYP51G1-Sc mRNA, suggesting a possible role of this transient biosynthetic intermediate as a bioactive signaling lipid molecule. Furthermore, treatments of leaves with 14C-labeled obtusifoliol demonstrated that this sterol could be transported in distal parts of the plant away from the sprayed leaves. Arabidopsis (Arabidopsis thaliana) CYP51 homozygous knockout mutants were also lethal, suggesting important roles for this enzymatic step and its substrate in plant development.
TL;DR: Two recent reports have shown that the BZR1-BES1 family of proteins is a class of novel plant-specific transcription factors that directly bind to and regulate BR-responsive genes, thus filling in key details of the link between hormonal signal transmission in the cytoplasm and transcriptional status change in the nucleus.