Transcription Factors BES1 Research Articles

BES1 directly binds to the promoter of the ACC oxidase 1 gene to regulate gravitropic response in the roots of Arabidopsis thaliana

ABSTRACTBrassinosteroids (BRs) are known to be endogenous regulators of ethylene production, suggesting that some BR activity in plant growth and development is associated with ethylene. Here, we demonstrated that ethylene production in Arabidopsis thaliana roots is increased by BR signaling via the ethylene biosynthetic gene for ACC oxidase 1 (ACO1). Electrophoretic mobility shift and chromatin immune-precipitation assays showed that the BR transcription factor BES1 directly binds to two E-box sequences located in the intergenic region of ACO1. GUS expression using site mutations of the E-box sequences verified that ACO1 is normally expressed only when BES1 binds to the E-boxes in the putative promoter of ACO1, indicating that this binding is essential for ACO1 expression and the subsequent production of ethylene in A. thaliana roots. BR exogenously applied to A. thaliana roots enhanced the gravitropic response. Additionally, bes1-D exhibited a greater gravitropic response than did the wild-type specimens, proving that BR is a positive regulator of the gravitropic response in A. thaliana roots. The knock-down mutant aco1-1 showed a slightly lower gravitropic response than did the wild-type specimens, while bes1-D X aco1-1 exhibited a lower gravitropic response than did bes1-D. Therefore, ACO1 is a direct downstream target for BR transcription factor BES1, which controls ethylene production for gravitropism in A. thaliana roots.

Active BR signalling adjusts the subcellular localisation of BES1/HSP90 complex formation.

Heat shock proteins 90 (HSP90) are essential and play critical roles in the adaptation of organisms to diverse stimuli. In plants, HSP90 are involved in auxin, jasmonate and brassinosteroid (BR) signalling pathways. The BR-promoted activation of the BES1 transcription factor regulates BR-responsive genes. Using genetic, physiological, fluorescence live cell imaging, molecular and biochemical approaches, such as phenotypic analysis, co-immunoprecipitation assay, yeast-two hybrid and Bimolecular fluorescence complementation (BiFC), we studied complex formation between BES1 and HSP90 under control conditions and active BR signalling. Further, we determined the effect of the pharmacological inhibition of HSP90 ATPase activity on hypocotyl elongation of bes1-D mutant. We determined that HSP90 interact with BES1 in the nucleus and in the cytoplasm. During active BR signalling, nuclear complexes were absent while cytoplasmic HSP90/BES1 complexes were prominent. Our results showed that the hypocotyl length of bes1-D mutants was highly reduced when HSP90 was challenged by the geldanamycin (GDA) inhibitor of the ATPase activity of HSP90. Active BR signalling could not rescue the GDA effect on the hypocotyl elongation of bes1-D. Our results reveal that the constitutively active BES1 in the bes1-D mutant is hypersensitive to GDA. The interaction of HSP90 with BES1 argues that HSP90 facilitate the nuclear metastable conformation of BES1 to regulate BR-dependent gene expression, and our data show that HSP90 assist in the compartmentalised cycle of BES1 during active BR signalling.

Auxin promotion of seedling growth via ARF5 is dependent on the brassinosteroid-regulated transcription factors BES1 and BEH4.

Seedlings must continually calibrate their growth in response to the environment. Auxin and brassinosteroids (BRs) are plant hormones that work together to control growth responses during photomorphogenesis. We used our previous analysis of promoter architecture in an auxin and BR target gene to guide our investigation into the broader molecular bases and biological relevance of transcriptional co‐regulation by these hormones. We found that the auxin‐regulated transcription factor Auxin Responsive Factor 5 (ARF5) and the brassinosteroid‐regulated transcription factor BRI1‐EMS‐Suppressor 1/Brassinazole Resistant 2 (BES1) co‐regulated a subset of growth‐promoting genes via conserved bipartite cis‐regulatory elements. Moreover, ARF5 binding to DNA could be enriched by increasing BES1 levels. The evolutionary loss of bipartite elements in promoters results in loss of hormone responsiveness. We also identified another member of the BES1/BZR1 family called BEH4 that acts partially redundantly with BES1 to regulate seedling growth. Double mutant analysis showed that BEH4 and not BZR1 were required alongside BES1 for normal auxin response during early seedling development. We propose that an ARF5‐BES1/BEH4 transcriptional module acts to promote growth via modulation of a diverse set of growth‐associated genes.

In silico genome-wide identification and comprehensive characterization of the BES1 gene family in soybean

The BES1 transcription factor family play a central role in brassinosteroid signaling pathway that regulates a wide range of plant growth and developmental processes, as well as resistances to various stresses. However, no comprehensive study of the BES1 gene family in soybean has been reported. In this work, 16 GmBES1-like genes were identified in soybean, which could be divided into two clades based on their phylogenetic relationships, gene structures and motif compositions. We then examined their duplication status and evolutionary models. The result showed that most of the GmBES1-like genes have duplicated counterparts generated from the recent Glycine WGD event, and these genes are originated from 6 distinct ancestors before the Gamma WGT event. We further studied the expression profiles of GmBES1-like genes, and found their spatio-temporal and stressed expression patterns varied tremendously. For example, GmBES1-5 and GmBES1-6 were highly expressed in almost every sample, whereas GmBES1-7 and GmBES1-8 were not expressed. Additionally, interaction network analysis revealed the presence of 3 clusters between GmBES1-like genes and other associated genes, implying that they have both the conserved and divergent functions. Lastly, we analyzed the genetic diversity of GmBES1-like genes in 302 resequenced wild, landrace and improved soybean accessions. It showed that most of these genes are well conserved, and they are not changed during domestication and improvement. These results provide insights into the characterization of GmBES1 family and lay the foundation for further functional study of such genes.

Genome-wide analysis of BES1 genes in Gossypium revealed their evolutionary conserved roles in brassinosteroid signaling.

Brassinosteroids (BRs), which are essential phytohormones for plant growth and development, are important for cotton fiber development. Additionally, BES1 transcription factors are critical for BR signal transduction. However, cotton BES1 family genes have not been comprehensively characterized. In this study, we identified 11 BES1 genes in G. arboreum, 11 in G. raimondii, 16 in G. barbadense, and 22 in G. hirsutum. The BES1 sequences were significantly conserved in the Arabidopsis thaliana, rice, and upland cotton genomes. A total of 94 BES1 genes from 10 different plant species were divided into three clades according to the neighbor-joining and minimum-evolution methods. Moreover, the exon/intron patterns and motif distributions were highly conserved among the A. thaliana and cotton BES1 genes. The collinearity among the orthologs from the At and Dt subgenomes was estimated. Segmental duplications in the At and Dt subgenomes were primarily responsible for the expansion of the cotton BES1 gene family. Of the GhBES1 genes, GhBES1.4_At/Dt exhibited BL-induced expression and was predominantly expressed in fibers. Furthermore, Col-0/mGhBES1.4_At plants produced curled leaves with long and bent petioles. These transgenic plants also exhibited decreased hypocotyl sensitivity to brassinazole and constitutive BR induced/repressed gene expression patterns. The constitutive BR responses of the plants overexpressing mGhBES1.4_At were similar to those of the bes1-D mutant.

PhyB Interacts with BES1 to Regulate Brassinosteroid Signaling in Arabidopsis.

Light is an important environmental factor, which mainly inhibits hypocotyl elongation through various photoreceptors. In contrast, brassinosteroids (BRs) are major hypocotyl elongation-promoting hormones in plants, which could optimize photomorphogenesis concurrent with external light. However, the precise molecular mechanisms underlying the antagonism of light and BR signaling remain largely unknown. Here we show that the Arabidopsis red light receptor phyB is involved in inhibition of BR signaling via its direct interaction with the BR transcription factor BES1. In our study, the phyB mutant displays BR hypersensitivity, which is repressed in transgenic plants overexpressing phyB, suggesting that phyB negatively regulates the BR signaling pathway. In addition, protein interaction results show that phyB directly interacts with dephosphorylated BES1, the physiologically active form of BES1 induced by BR, in a red light-dependent manner. Genetic analyses suggest that phyB may act partially through BES1 to regulate BR signaling. Transcriptomic data and quantitative real-time PCR assay further show that phyB-mediated red light inhibits BR signaling by repressing expression of BES1 target genes, including the BR biosynthesis genes DWF4, the SAUR family and the PRE family genes required for promoting cell elongation. Finally, we found that red light treatment inhibits the DNA-binding activity of BES1 and photoactivated phyB represses the transcriptional activity of BES1 under red light. Taken together, we suggest that the interaction of phyB with dephosphorylated BES1 may allow plants to balance light and BR signaling by repressing transcriptional activity of BES1 to regulate expression of its target genes.

Overview of human embryonic development

The BES1 transcription factor family play a central role in brassinosteroid signaling pathway that regulates a wide range of plant growth and developmental processes, as well as resistances to various stresses. However, no comprehensive study of the BES1 gene family in soybean has been reported. In this work, 16 GmBES1-like genes were identified in soybean, which could be divided into two clades based on their phylogenetic relationships, gene structures and motif compositions. We then examined their duplication status and evolutionary models. The result showed that most of the GmBES1-like genes have duplicated counterparts generated from the recent Glycine WGD event, and these genes are originated from 6 distinct ancestors before the Gamma WGT event. We further studied the expression profiles of GmBES1-like genes, and found their spatio-temporal and stressed expression patterns varied tremendously. For example, GmBES1-5 and GmBES1-6 were highly expressed in almost every sample, whereas GmBES1-7 and GmBES1-8 were not expressed. Additionally, interaction network analysis revealed the presence of 3 clusters between GmBES1-like genes and other associated genes, implying that they have both the conserved and divergent functions. Lastly, we analyzed the genetic diversity of GmBES1-like genes in 302 resequenced wild, landrace and improved soybean accessions. It showed that most of these genes are well conserved, and they are not changed during domestication and improvement. These results provide insights into the characterization of GmBES1 family and lay the foundation for further functional study of such genes.

Brassinosteroids regulate root growth by controlling reactive oxygen species homeostasis and dual effect on ethylene synthesis in Arabidopsis.

The brassinosteroids (BRs) represent a class of phytohormones, which regulate numerous aspects of growth and development. Here, a det2-9 mutant defective in BR synthesis was identified from an EMS mutant screening for defects in root length, and was used to investigate the role of BR in root development in Arabidopsis. The det2-9 mutant displays a short-root phenotype, which is result from the reduced cell number in root meristem and decreased cell size in root maturation zone. Ethylene synthesis is highly increased in the det2-9 mutant compared with the wild type, resulting in the hyper-accumulation of ethylene and the consequent inhibition of root growth. The short-root phenotype of det2-9 was partially recovered in the det2-9/acs9 double mutant and det2-9/ein3/eil1-1 triple mutant which have defects either in ethylene synthesis or ethylene signaling, respectively. Exogenous application of BR showed that BRs either positively or negatively regulate ethylene biosynthesis in a concentration-dependent manner. Different from the BR induced ethylene biosynthesis through stabilizing ACSs stability, we found that the BR signaling transcription factors BES1 and BZR1 directly interacted with the promoters of ACS7, ACS9 and ACS11 to repress their expression, indicating a native regulation mechanism under physiological levels of BR. In addition, the det2-9 mutant displayed over accumulated superoxide anions (O2-) compared with the wild-type control, and the increased O2- level was shown to contribute to the inhibition of root growth. The BR-modulated control over the accumulation of O2- acted via the peroxidase pathway rather than via the NADPH oxidase pathway. This study reveals an important mechanism by which the hormone cross-regulation between BRs and ethylene or/and ROS is involved in controlling root growth and development in Arabidopsis.

Arabidopsis G-Protein β Subunit AGB1 Interacts with BES1 to Regulate Brassinosteroid Signaling and Cell Elongation.

In Arabidopsis, brassinosteroids (BR) are major growth-promoting hormones, which integrate with the heterotrimeric guanine nucleotide-binding protein (G-protein) signals and cooperatively modulate cell division and elongation. However, the mechanisms of interaction between BR and G-protein are not well understood. Here, we show that the G-protein β subunit AGB1 directly interacts with the BR transcription factor BES1 in vitro and in vivo. An AGB1-null mutant, agb1-2, displays BR hyposensitivity and brassinazole (BRZ, BR biosynthesis inhibitor) hypersensitivity, which suggests that AGB1 positively mediates the BR signaling pathway. Moreover, we demonstrate that AGB1 synergistically regulates expression of BES1 target genes, including the BR biosynthesis genes CPD and DWF4 and the SAUR family genes required for promoting cell elongation. Further, Western blot analysis of BES1 phosphorylation states indicates that the interaction between AGB1 and BES1 alters the phosphorylation status of BES1 and increases the ratio of dephosphorylated to phosphorylated BES1, which leads to accumulation of dephosphorylated BES1 in the nucleus. Finally, AGB1 promotes BES1 binding to BR target genes and stimulates the transcriptional activity of BES1. Taken together, our results demonstrate that AGB1 positively regulates cell elongation by affecting the phosphorylation status and transcriptional activity of BES1.

TTL Proteins Scaffold Brassinosteroid Signaling Components at the Plasma Membrane to Optimize Signal Transduction in Plant Cells

Brassinosteroids (BRs) form a group of steroidal hormones essential for plant growth, development and stress responses. BR perception at the plasma membrane initiates a series of phosphorylation events enabling the nuclear accumulation and activity of the key transcription factors BZR1 and BES1. Previous studies support that all BR signaling components form an interconnected pathway, although it is not known how these proteins are brought together for the prompt signal transduction from the plasma membrane to the nucleus. Here we report that plant-specific Tetratricopeptide Thioredoxin- Like (TTL) proteins are positive regulators of BR signaling that function as scaffold for the BR signaling components in Arabidopsis. TTL3 directly interacts with BZR1 and the phosphatase BSU1, and associates in vivo with most core components involved in BR signaling, with the exception of the BAK1 coreceptor. TTL3 is mainly localized in the cytoplasm, and BR treatment increases its localization at the plasma membrane. In addition, the expression of TTL3 strengthens the association of BR-signaling components BSK1 and BZR1 at the plasma membrane. Consistent with a role in BR signaling, mutations in TTL3, and related TTL1 and TTL4 genes cause reduced BR responses, and these defects that highly enhanced in a triple ttl1 ttl3 ttl4 mutant. We propose a novel mechanistic model for BR signaling, in which cytoplasmic/nuclear BR components bound to TTL proteins are recruited to the plasma membrane upon BR perception, which in turn allows the assembly of a BR signaling complex with the goal of ensuring dephosphorylation and nuclear accumulation of the transcription factors BZR1 and BES1. Interestingly, this novel TTL scaffold model for BR signaling resembles that of Wnt signaling in metazoans, in which TTL proteins would act similar to Axin1, optimizing signaling efficiency of the cascade by promoting the assembly of the signaling complex at the plasma membrane.

WRKY transcription factors are involved in brassinosteroid signaling and mediate the crosstalk between plant growth and drought tolerance

ABSTRACTBrassinosteroids (BRs) are critical for the plant growth and development. BRs signal through the plasma membrane localized receptor-like kinases to downstream transcription factors BES1/BZR1 to regulate the expression of thousands of genes for various BR responses. In addition to the role in plant growth and development, BRs have been implicated in responses to environmental stresses such as drought. However, the mechanism through which BRs regulate drought have just begun emerging. We have recently found that a group of WRKY transcription factors, WRKY46, WRKY54, WRKY70, which are well known for the function in abiotic and biotic stress, cooperates with BES1 to mediate BR-regulated drought response. The wrky46 wrky54 wrky70 triple mutants showed growth defect, likely due to impaired BR signaling as well as some reduction of endogenous BR level. WRKY46/54/70 cooperates with BES1 to regulate the expression of BR target genes to promote growth. We also found that WRKY46/54/70 negatively modulates drought tolerance by globally repressing drought-inducible gene expression. Thus, our result uncovers a new role for WRKY transcription factors in BR signaling and provides the molecular mechanism for BR-regulated plant growth and drought stress through WRKY46/54/70 and BES1 transcription factors.

BZR1 Positively Regulates Freezing Tolerance via CBF-Dependent and CBF-Independent Pathways in Arabidopsis

Cold stress is a major environmental factor that adversely affects plant growth and development. The C-repeat binding factor/DRE binding factor 1 (CBF/DREB1) transcriptional regulatory cascade has been shown to play important roles in plant response to cold. Here we demonstrate that two key components of brassinosteroid (BR) signaling modulate freezing tolerance of Arabidopsis plants. The loss-of-function mutant of the GSK3-like kinases involved in BR signaling, bin2-3 bil1 bil2, showed increased freezing tolerance, whereas overexpression of BIN2 resulted in hypersensitivity to freezing stress under both non-acclimated and acclimated conditions. By contrast, gain-of-function mutants of the transcription factors BZR1 and BES1 displayed enhanced freezing tolerance, and consistently cold treatment could induce the accumulation of dephosphorylated BZR1. Biochemical and genetic analyses showed that BZR1 acts upstream of CBF1 and CBF2 to directly regulate their expression. Moreover, we found that BZR1 also regulated other COR genes uncoupled with CBFs, such as WKRY6, PYL6, SOC1, JMT, and SAG21, to modulate plant response to cold stress. Consistently, wrky6 mutants showed decreased freezing tolerance. Taken together, our results indicate that BZR1 positively modulates plant freezing tolerance through CBF-dependent and CBF-independent pathways.

BES1 regulates the localization of the brassinosteroid receptor BRL3 within the provascular tissue of the Arabidopsis primary root.

Brassinosteroid (BR) hormones are important regulators of plant growth and development. Recent studies revealed the cell-specific role of BRs in vascular and stem cell development by the action of cell-specific BR receptor complexes and downstream signaling components in Arabidopsis thaliana Despite the importance of spatiotemporal regulation of hormone signaling in the control of plant vascular development, the mechanisms that confer cellular specificity to BR receptors within the vascular cells are not yet understood. The present work shows that BRI1-like receptor genes 1 and 3 (BRL1 and BRL3) are differently regulated by BRs. By using promoter deletion constructs of BRL1 and BRL3 fused to GFP/GUS (green fluorescent protein/β-glucuronidase) reporters in Arabidopsis, analysis of their cell-specific expression and regulation by BRs in the root apex has been carried out. We found that BRL3 expression is finely modulated by BRs in different root cell types, whereas the location of BRL1 appears to be independent of this hormone. Physiological and genetic analysis show a BR-dependent expression of BRL3 in the root meristem. In particular, BRL3 expression requires active BES1, a central transcriptional effector within the BRI1 pathway. ChIP analysis showed that BES1 directly binds to the BRRE present in the BRL3 promoter region, modulating its transcription in different subsets of cells of the root apex. Overall our study reveals the existence of a cell-specific negative feedback loop from BRI1-mediated BES1 transcription factor to BRL3 in phloem cells, while contributing to a general understanding of the spatial control of steroid signaling in plant development.

Genome-wide analysis of the BES1 transcription factor family in Chinese cabbage (Brassica rapa ssp. pekinensis)

The BES1 transcription factor family is one of the most important families involved in growth and development, hormone responses, and abiotic or biotic stress responses in plants. Completion of the sequencing of the Brassica rapa genome enabled us to undertake a genome-wide identification and functional study of the gene families related to the morphological diversity and agronomic traits of Brassica crops. Therefore, systematic analysis of the BES1 gene family in Chinese cabbage has profound significance. In this study, 15 BrBES1 genes were identified in Chinese cabbage, which could be classified into two groups according to their gene structural characteristics and motif compositions obtained. Fifteen BrBES1 genes mapped on chromosomes were further assigned to three subgenomes and eight ancestral karyotypes. Distribution mapping shows that BES1 proteins are non-randomly localized in eight chromosomes. Fourteen orthologous gene pairs are shared by Chinese cabbage and Arabidopsis. The expansion of BES1 genes in Chinese cabbage results from genome triplication. Quantitative RT-PCR analysis showed that expressions of BrBES1 genes varied widely under different abiotic stress treatments for different times. Numerous stress–responsive candidate genes were obtained. Our systematic analyses provided insights into the characterization of the BES1 genes in Chinese cabbage and basis for further functional studies of such genes.

BRI1-EMS-suppressor 1 gain-of-function mutant shows higher susceptibility to necrotrophic fungal infection

Brassinosteroids (BRs) are plant-specific steroids that are involved in plant growth and defense responses. However, the exact roles of BR in plant defense are unclear. We used the bes1-D gain-of-function mutant to define the underlying relationship between plant growth and defense through BR signaling and innate immunity. In bes1-D, further downstream component BES1 transcription factor is stabilized, leading to the activation of BR signaling. Previous reports on BES1 target genes showed that approximately 10% are related to biotic stress responses. Therefore, the bes1-D PTI responses were examined. The bes1-D mutant was specifically susceptible to Alternaria brassicicola, a necrotrophic fungus, which successfully produced spore, resulting in considerable cell death. However, it was not affected by a biotrophic pathogen, Pseudomonas syringae pv. tomato (Pst) DC3000. Instead of a ROS burst, a representative initial PTI responses, higher ROS accumulation was sustained in bes1-D than in the wild type plant. PDF1.2 expression was not induced in response to fungal pathogen infection in bes1-D. These results suggest that BES1 is also involved in JA-related defense responses, especially in response to necrotrophic pathogens.

OCTOPUS Negatively Regulates BIN2 to Control Phloem Differentiation in Arabidopsis thaliana

The phloem is a vascular strand that conducts photoassimilates and systemic signals throughout the plant to coordinate growth. To date, few molecular genetic determinants have been identified to control both specification and differentiation of this tissue [1-3]. Among them, OCTOPUS (OPS) protein was previously identified as a polarly localized plasma membrane-associated protein of unknown biochemical function whose broad provascular expression becomes restricted to the phloem upon differentiation [2]. OPS loss-of-function mutants showed an altered vascular network in cotyledons and an intermittent phloem differentiation in the root [2, 4]. Here, we demonstrate a role for OPS as a positive regulator of the brassinosteroid (BR) signaling pathway. Indeed, transgenic lines overexpressing OPS (OPS-OE) display the hallmarks of constitutively overactivated BR mutants. Physiological and genetic analyses place OPS as a positive regulator of the BR signaling pathway upstream of the key transcription factors BES1 and BZR1. Directed protein interactions with known BR signaling proteins identified BIN2, a GSK3 protein involved in multiple signaling pathways, as a partner of OPS. This interaction recruits BIN2 to the plasma membrane, thus preventing its inhibitory activity in the nucleus. Finally, both bikinin (a potent inhibitor of GSK3 [5]) treatment and downstream dominant mutants bes1-D [6] and bzr1-D [7] can rescue phloem defects of ops in the root. Together, our data show that OPS antagonizes BIN2 to promote phloem differentiation.

Brassinosteroids Are Master Regulators of Gibberellin Biosynthesis in Arabidopsis.

Plant growth and development are highly regulated processes that are coordinated by hormones including the brassinosteroids (BRs), a group of steroids with structural similarity to steroid hormones of mammals. Although it is well understood how BRs are produced and how their signals are transduced, BR targets, which directly confer the hormone's growth-promoting effects, have remained largely elusive. Here, we show that BRs regulate the biosynthesis of gibberellins (GAs), another class of growth-promoting hormones, in Arabidopsis thaliana. We reveal that Arabidopsis mutants deficient in BR signaling are severely impaired in the production of bioactive GA, which is correlated with defective GA biosynthetic gene expression. Expression of the key GA biosynthesis gene GA20ox1 in the BR signaling mutant bri1-301 rescues many of its developmental defects. We provide evidence that supports a model in which the BR-regulated transcription factor BES1 binds to a regulatory element in promoters of GA biosynthesis genes in a BR-induced manner to control their expression. In summary, our study underscores a role of BRs as master regulators of GA biosynthesis and shows that this function is of major relevance for the growth and development of vascular plants.

Down-regulation of BdBRI1, a putative brassinosteroid receptor gene produces a dwarf phenotype with enhanced drought tolerance in Brachypodium distachyon

Brassinosteroids (BRs) play important roles in plant growth, development and responses to a range of environmental cues. Although the mechanism of how BRs regulate growth and development is well-understood in Arabidopsis, the effect of BRs on stress tolerance, particularly drought tolerance remains unknown. We isolated a BRI1 (BRASSINOSTEROID INSENSITIVE 1) homologous gene, BdBRI1 from Brachypodium distachyon, a model for temperate grasses and cereals, created and characterized RNA interference (RNAi) knockdown mutants for BdBRI1 in Brachypodium. The loss-of-function BdBRI1-RNAi mutants exhibited reduced plant height, shortened internodes, narrow and short leaf, and reduced expression of BR signaling genes, BdBES1, BdBZR1, BdBLE2, and enhanced expression of BR biosynthesis genes BdD2, BdCPD and BdDWF4. More importantly, BdBRI1 RNAi mutants exhibited enhanced drought tolerance, accompanied by highly elevated expression of drought-responsive genes, BdP5CS, BdCOR47/BdRD17, together with BdERD1 and BdRD26, two putative targets of the transcription factors BES1 and BZR1 that are key components of the BR signaling pathway. Our results suggest that BR signaling and biosynthesis are largely conserved among Arabidopsis, rice and Brachypodium, and that BR signaling plays an important role in drought tolerance by directly regulating expression of key drought-responsive genes. The effect of BR biosynthesis or crosstalks between BR and other hormones or components of stress signaling pathways on drought tolerance is discussed.

Turning on the microscope turret: a new view for the study of brassinosteroid signaling in plant development

Brassinosteroid (BR) hormones are essential for plant growth and development. In Arabidopsis, the general understanding of BR signaling has been greatly attained by genetic and biochemical approaches that led to the identification of central BR signaling components, from the BRI1 receptor at the plasma membrane to downstream acting BR-regulated BRZ1 and BES1 transcription factors in the nuclei. Recently, an emerging trend is being established to further advance our understanding of the BR signaling pathway in plant development. Scientists have turned on the microscope lens turret to revisit the pleiotropic phenotypes of the BR mutants at a higher magnification, uncovering novel and specific cellular defects in the plant. In-depth phenotypic analysis in combination with the search for cell-specific signaling components that are responsible for those particular defects in the mutants are leading to: (1) definition of novel roles for BRs in vascular development, (2) unraveling BR function in cell division through quantitative analysis of Arabidopsis root growth, (3) establishment of a molecular connection between known patterning and BR-signaling components in organ boundary and stomata development and (4) development of novel strategies toward the identification of BR signaling components with spatiotemporal resolution. In this review, we highlight the importance of these emerging studies to investigate the spatiotemporal control of BR pathways in plant development.

The Protein Chaperone HSP90 Can Facilitate the Divergence of Gene Duplicates

The heat-shock protein 90 (HSP90) acts as a chaperone by ensuring proper maturation and folding of its client proteins. The HSP90 capacitor hypothesis holds that interactions with HSP90 allow proteins to accumulate mutations while maintaining function. Following this logic, HSP90 clients would be predicted to show relaxed selection compared with nonclients. In this study, we identify a new HSP90 client in the plant steroid hormone pathway: the transcription factor BES1. Its closest paralog, BZR1, is not an HSP90 client. This difference in HSP90 client status in two highly similar proteins enabled a direct test of the capacitor hypothesis. We find that BES1 shows relaxed selection compared to BZR1, hallmarks of neo- and subfunctionalization, and dynamic HSP90 client status across independent evolutionary paths. These results suggested that HSP90's influence on gene evolution may be detectable if we compare gene duplicates because duplicates share most other properties influencing evolutionary rate that might otherwise conceal the chaperone's effect. We test this hypothesis using systematically identified HSP90 clients in yeast and observe a significant trend of HSP90 clients evolving faster than their nonclient paralogs. This trend was not detected when yeast clients and nonclients were compared without considering paralog status. Our data provide evidence that HSP90 influences selection on genes encoding its clients and facilitates divergence between gene duplicates.