SMART MODE:

NORMAL GENOMIC

• Huang D et al.
• A Gibberellin-Mediated DELLA-NAC Signaling Cascade Regulates Cellulose Synthesis in Rice.
• Plant Cell. 2015; 27: 1681-96
• Display abstract

• Cellulose, which can be converted into numerous industrial products, has important impacts on the global economy. It has long been known that cellulose synthesis in plants is tightly regulated by various phytohormones. However, the underlying mechanism of cellulose synthesis regulation remains elusive. Here, we show that in rice (Oryza sativa), gibberellin (GA) signals promote cellulose synthesis by relieving the interaction between SLENDER RICE1 (SLR1), a DELLA repressor of GA signaling, and NACs, the top-layer transcription factors for secondary wall formation. Mutations in GA-related genes and physiological treatments altered the transcription of CELLULOSE SYNTHASE genes (CESAs) and the cellulose level. Multiple experiments demonstrated that transcription factors NAC29/31 and MYB61 are CESA regulators in rice; NAC29/31 directly regulates MYB61, which in turn activates CESA expression. This hierarchical regulation pathway is blocked by SLR1-NAC29/31 interactions. Based on the results of anatomical analysis and GA content examination in developing rice internodes, this signaling cascade was found to be modulated by varied endogenous GA levels and to be required for internode development. Genetic and gene expression analyses were further performed in Arabidopsis thaliana GA-related mutants. Altogether, our findings reveal a conserved mechanism by which GA regulates secondary wall cellulose synthesis in land plants and provide a strategy for manipulating cellulose production and plant growth.

• Xu K et al.
• OsGRAS23, a rice GRAS transcription factor gene, is involved in drought stress response through regulating expression of stress-responsive genes.
• BMC Plant Biol. 2015; 15: 141-141
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• BACKGROUND: Drought is a major abiotic stress factors that reduces agricultural productivity. GRAS transcription factors are plant-specific proteins that play diverse roles in plant development. However, the functions of a number of GRAS genes identified in rice are unknown, especially the GRAS genes related to rice drought resistance have not been characterized. RESULTS: In this study, a novel GRAS transcription factor gene named OsGRAS23, which is located in a drought-resistant QTL interval on chromosome 4 of rice, was isolated. The expression of OsGRAS23 was induced by drought, NaCl, and jasmonic acid treatments. The OsGRAS23-GFP fused protein was localized in the nucleus of tobacco epidermal cells. A trans-activation assay in yeast cells demonstrated that the OsGRAS23 protein possessed a strong transcriptional activation activity. OsGRAS23-overexpressing rice plants showed improved drought resistance and oxidative stress tolerance as well as less H2O2 accumulation compared with the wild-type plants. Furthermore, microarray analysis showed that several anti-oxidation related genes were up-regulated in the OsGRAS23-overexpressing rice plants. The yeast one hybrid test indicated that OsGRAS23 could bind to the promoters of its potential target genes. CONCLUSIONS: Our results demonstrate that OsGRAS23 encodes a stress-responsive GRAS transcription factor and positively modulates rice drought tolerance via the induction of a number of stress-responsive genes.

• Nelis S, Conti L, Zhang C, Sadanandom A
• A functional Small Ubiquitin-like Modifier (SUMO) interacting motif (SIM) in the gibberellin hormone receptor GID1 is conserved in cereal crops and disrupting this motif does not abolish hormone dependency of the DELLA-GID1 interaction.
• Plant Signal Behav. 2015; 10: 987528-987528
• Display abstract

• Plants survive adversity by modulating their growth in response to changing environmental signals. The phytohormone Gibberellic acid (GA) plays a central role in regulating these adaptive responses by stimulating the degradation of growth repressing DELLA proteins which accumulate during stress. The current model for GA signaling describes how this hormone binds to its receptor GID1 so promoting association of GID1 with DELLA, which then undergoes ubiquitin-mediated proteasomal degradation. Recent data revealed that conjugation of DELLAs to the Small Ubiquitin-like Modifier (SUMO) protein enables plants to modulate its abundance during environmental stress. This is achieved by SUMOylated DELLAs sequestering GID1 via its SUMO interacting motif (SIM) allowing non-SUMOylated DELLAs to accumulate leading to growth restraint under stress and potential yield loss. We demonstrate that GID1 proteins across the major cereal crops contain a functional SIM able to bind SUMO1. Site directed mutagenesis and yeast 2 hybrid experiments reveal that it is possible to disrupt the SIM-SUMO interaction motif without affecting the GA dependent DELLA-GID1 interaction and thereby uncoupling SUMO-mediated inhibition from DELLA degradation. Arabidopsis plants overexpressing a SIM mutant allele of GID1 perform better at relieving DELLA restraint than wild-type GID1. This evidence suggests that manipulating the SIM motif in the GA receptor may provide a possible route to developing stress tolerant crops plants.

• Fonseca S, Rosado A, Vaughan-Hirsch J, Bishopp A, Chini A
• Molecular locks and keys: the role of small molecules in phytohormone research.
• Front Plant Sci. 2014; 5: 709-709
• Display abstract

• Plant adaptation, growth and development rely on the integration of many environmental and endogenous signals that collectively determine the overall plant phenotypic plasticity. Plant signaling molecules, also known as phytohormones, are fundamental to this process. These molecules act at low concentrations and regulate multiple aspects of plant fitness and development via complex signaling networks. By its nature, phytohormone research lies at the interface between chemistry and biology. Classically, the scientific community has always used synthetic phytohormones and analogs to study hormone functions and responses. However, recent advances in synthetic and combinational chemistry, have allowed a new field, plant chemical biology, to emerge and this has provided a powerful tool with which to study phytohormone function. Plant chemical biology is helping to address some of the most enduring questions in phytohormone research such as: Are there still undiscovered plant hormones? How can we identify novel signaling molecules? How can plants activate specific hormone responses in a tissue-specific manner? How can we modulate hormone responses in one developmental context without inducing detrimental effects on other processes? The chemical genomics approaches rely on the identification of small molecules modulating different biological processes and have recently identified active forms of plant hormones and molecules regulating many aspects of hormone synthesis, transport and response. We envision that the field of chemical genomics will continue to provide novel molecules able to elucidate specific aspects of hormone-mediated mechanisms. In addition, compounds blocking specific responses could uncover how complex biological responses are regulated. As we gain information about such compounds we can design small alterations to the chemical structure to further alter specificity, enhance affinity or modulate the activity of these compounds.

• Xue XY et al.
• Interaction between two timing microRNAs controls trichome distribution in Arabidopsis.
• PLoS Genet. 2014; 10: 1004266-1004266
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• The miR156-targeted squamosa promoter binding protein like (SPL) transcription factors function as an endogenous age cue in regulating plant phase transition and phase-dependent morphogenesis, but the control of SPL output remains poorly understood. In Arabidopsis thaliana the spatial pattern of trichome is a hallmark of phase transition and governed by SPLs. Here, by dissecting the regulatory network controlling trichome formation on stem, we show that the miR171-targeted lost meristems 1 (LOM1), LOM2 and LOM3, encoding GRAS family members previously known to maintain meristem cell polarity, are involved in regulating the SPL activity. Reduced LOM abundance by overexpression of miR171 led to decreased trichome density on stems and floral organs, and conversely, constitutive expression of the miR171-resistant LOM (rLOM) genes promoted trichome production, indicating that LOMs enhance trichome initiation at reproductive stage. Genetic analysis demonstrated LOMs shaping trichome distribution is dependent on SPLs, which positively regulate trichome repressor genes TRICHOMELESS 1 (TCL1) and TRIPTYCHON (TRY). Physical interaction between the N-terminus of LOMs and SPLs underpins the repression of SPL activity. Importantly, other growth and developmental events, such as flowering, are also modulated by LOM-SPL interaction, indicating a broad effect of the LOM-SPL interplay. Furthermore, we provide evidence that MIR171 gene expression is regulated by its targeted LOMs, forming a homeostatic feedback loop. Our data uncover an antagonistic interplay between the two timing miRNAs in controlling plant growth, phase transition and morphogenesis through direct interaction of their targets.

• Locascio A, Blazquez MA, Alabadi D
• Dynamic regulation of cortical microtubule organization through prefoldin-DELLA interaction.
• Curr Biol. 2013; 23: 804-9
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• Plant morphogenesis relies on specific patterns of cell division and expansion. It is well established that cortical microtubules influence the direction of cell expansion, but less is known about the molecular mechanisms that regulate microtubule arrangement. Here we show that the phytohormones gibberellins (GAs) regulate microtubule orientation through physical interaction between the nuclear-localized DELLA proteins and the prefoldin complex, a cochaperone required for tubulin folding. In the presence of GA, DELLA proteins are degraded, and the prefoldin complex stays in the cytoplasm and is functional. In the absence of GA, the prefoldin complex is localized to the nucleus, which severely compromises alpha/beta-tubulin heterodimer availability, affecting microtubule organization. The physiological relevance of this molecular mechanism was confirmed by the observation that the daily rhythm of plant growth was accompanied by coordinated oscillation of DELLA accumulation, prefoldin subcellular localization, and cortical microtubule reorientation.

• Band LR, Bennett MJ
• Mapping the site of action of the Green Revolution hormone gibberellin.
• Proc Natl Acad Sci U S A. 2013; 110: 4443-4

• Sarnowska EA et al.
• DELLA-interacting SWI3C core subunit of switch/sucrose nonfermenting chromatin remodeling complex modulates gibberellin responses and hormonal cross talk in Arabidopsis.
• Plant Physiol. 2013; 163: 305-17
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• Switch (SWI)/Sucrose Nonfermenting (SNF)-type chromatin-remodeling complexes (CRCs) are involved in regulation of transcription, DNA replication and repair, and cell cycle. Mutations of conserved subunits of plant CRCs severely impair growth and development; however, the underlying causes of these phenotypes are largely unknown. Here, we show that inactivation of SWI3C, the core component of Arabidopsis (Arabidopsis thaliana) SWI/SNF CRCs, interferes with normal functioning of several plant hormone pathways and alters transcriptional regulation of key genes of gibberellin (GA) biosynthesis. The resulting reduction of GA4 causes severe inhibition of hypocotyl and root elongation, which can be rescued by exogenous GA treatment. In addition, the swi3c mutation inhibits DELLA-dependent transcriptional activation of GIBBERELLIN-INSENSITIVE DWARF1 (GID1) GA receptor genes. Down-regulation of GID1a in parallel with the DELLA repressor gene REPRESSOR OF GA1-3 1 in swi3c indicates that lack of SWI3C also leads to defects in GA signaling. Together with the recent demonstration of function of SWI/SNF ATPase BRAHMA in the GA pathway, these results reveal a critical role of SWI/SNF CRC in the regulation of GA biosynthesis and signaling. Moreover, we demonstrate that SWI3C is capable of in vitro binding to, and shows in vivo bimolecular fluorescence complementation interaction in cell nuclei with, the DELLA proteins RGA-LIKE2 and RGA-LIKE3, which affect transcriptional activation of GID1 and GA3ox (GIBBERELLIN 3-OXIDASE) genes controlling GA perception and biosynthesis, respectively. Furthermore, we show that SWI3C also interacts with the O-GlcNAc (O-linked N-acetylglucosamine) transferase SPINDLY required for proper functioning of DELLAs and acts hypostatically to (SPINDLY) in the GA response pathway. These findings suggest that DELLA-mediated effects in GA signaling as well as their role as a hub in hormonal cross talk may be, at least in part, dependent on their direct physical interaction with complexes responsible for modulation of chromatin structure.

• Golldack D, Li C, Mohan H, Probst N
• Gibberellins and abscisic acid signal crosstalk: living and developing under unfavorable conditions.
• Plant Cell Rep. 2013; 32: 1007-16
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• Plants adapt to adverse environments by integrating growth and development to environmentally activated cues. Within the adaptive signaling networks, plant hormones tightly control convergent developmental and stress adaptive processes and coordinate cellular responses to external and internal conditions. Recent studies have uncovered novel antagonizing roles of the plant hormones gibberellin (GA) and abscisic acid (ABA) in integrating growth and development in plants with environmental signaling. According to current concepts, GRAS transcription factors of the DELLA and SCARECROW-LIKE (SCL) types have a key role as major growth regulators and have pivotal functions in modulating GA signaling. Significantly, current models emphasize a function of DELLA proteins as central regulators in GA homeostasis. DELLA proteins interact with the cellular GA receptor GID1 (GA-INSENSITIVE DWARF1) and degradation of DELLAs activates the function of GA. Supplementary to the prevailing view of a pivotal role of GRAS family transcriptional factors in plant growth regulation, recent work has suggested that the DELLA and SCL proteins integrate generic GA responses into ABA-controlled abiotic stress tolerance. Here, we review and discuss how GRAS type proteins influence plant development and versatile adaptation as hubs in GA and ABA triggered signaling pathways.

• Dixit R
• Plant cytoskeleton: DELLA connects gibberellins to microtubules.
• Curr Biol. 2013; 23: 47981-47981
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• A new study reveals that DELLA proteins directly interact with the prefoldin complex, thus regulating tubulin subunit availability in a gibberellin-dependent manner. This finding provides a mechanistic link between the growth-promoting plant hormone gibberellin and cortical microtubule organization.

• Middleton AM et al.
• Mathematical modeling elucidates the role of transcriptional feedback in gibberellin signaling.
• Proc Natl Acad Sci U S A. 2012; 109: 7571-6
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• The hormone gibberellin (GA) is a key regulator of plant growth. Many of the components of the gibberellin signal transduction [e.g., GIBBERELLIN INSENSITIVE DWARF 1 (GID1) and DELLA], biosynthesis [e.g., GA 20-oxidase (GA20ox) and GA3ox], and deactivation pathways have been identified. Gibberellin binds its receptor, GID1, to form a complex that mediates the degradation of DELLA proteins. In this way, gibberellin relieves DELLA-dependent growth repression. However, gibberellin regulates expression of GID1, GA20ox, and GA3ox, and there is also evidence that it regulates DELLA expression. In this paper, we use integrated mathematical modeling and experiments to understand how these feedback loops interact to control gibberellin signaling. Model simulations are in good agreement with in vitro data on the signal transduction and biosynthesis pathways and in vivo data on the expression levels of gibberellin-responsive genes. We find that GA-GID1 interactions are characterized by two timescales (because of a lid on GID1 that can open and close slowly relative to GA-GID1 binding and dissociation). Furthermore, the model accurately predicts the response to exogenous gibberellin after a number of chemical and genetic perturbations. Finally, we investigate the role of the various feedback loops in gibberellin signaling. We find that regulation of GA20ox transcription plays a significant role in both modulating the level of endogenous gibberellin and generating overshoots after the removal of exogenous gibberellin. Moreover, although the contribution of other individual feedback loops seems relatively small, GID1 and DELLA transcriptional regulation acts synergistically with GA20ox feedback.

• Gaiji N, Cardinale F, Prandi C, Bonfante P, Ranghino G
• The computational-based structure of Dwarf14 provides evidence for its role as potential strigolactone receptor in plants.
• BMC Res Notes. 2012; 5: 307-307
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• BACKGROUND: Strigolactones (SLs) are recently identified plant hormones modulating root and shoot branching. Besides their endogenous role within the producing organism, SLs are also key molecules in the communication of plants with arbuscular mycorrhizal (AM) fungi and parasitic weeds. In fact SLs are exuded into the rhizosphere where they act as a host-derived signal, stimulating the germination of the seeds of parasitic plants which would not survive in the absence of a host root to colonize. Similarly, their perception by AM fungi causes extensive hyphal branching; this is a prerequisite for effective root colonization, since it increases the number of potential contact points with the host surface. In spite of the crucial and multifaceted biological role of SLs, there is no information on the receptor(s) which bind(s) such active molecules, neither in the producing plants, or in parasitic weeds or AM fungi. RESULTS: In this work, we applied homology modelling techniques to investigate the structure of the protein encoded by the gene Dwarf14, which was first identified in rice as conferring SLs insensitivity when mutated. The best sequence identity was with bacterial RsbQ. Both proteins belong to the superfamily of alpha/beta-fold hydrolases, some members of which play a role in the metabolism or signalling of plant hormones. The Dwarf14 (D14) structure was refined by means of molecular dynamics simulations. In order to support the hypothesis that D14 could be an endogenous SLs receptor, we performed docking experiments with a natural ligand. CONCLUSIONS: It is suggested that D14 interacts with and thereby may act as a receptor for SLs in plants. This hypothesis offers a starting point to experimentally study the mechanism of its activity in vivo by means of structural, molecular and genetic approaches. Lastly, knowledge of the putative receptor structure will boost the research on analogues of the natural substrates as required for agricultural applications.

• Tanimoto E
• Tall or short? Slender or thick? A plant strategy for regulating elongation growth of roots by low concentrations of gibberellin.
• Ann Bot. 2012; 110: 373-81
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• BACKGROUND: Since the plant hormone gibberellin (GA) was discovered as a fungal toxin that caused abnormal elongation of rice shoots, the physiological function of GA has mainly been investigated in relation to the regulation of plant height. However, an indispensable role for GA in root growth has been elucidated by using severely GA-depleted plants, either with a gene mutation in GA biosynthesis or which have been treated by an inhibitor of GA biosynthesis. The molecular sequence of GA signalling has also been studied to understand GA functions in root growth. SCOPE: This review addresses research progress on the physiological functions of GA in root growth. Concentration-dependent stimulation of elongation growth by GA is important for the regulation of plant height and root length. Thus the endogenous level of GA and/or the GA sensitivity of shoots and roots plays a role in determining the shoot-to-root ratio of the plant body. Since the shoot-to-root ratio is an important parameter for agricultural production, control of GA production and GA sensitivity may provide a strategy for improving agricultural productivity. The sequence of GA signal transduction has recently been unveiled, and some component molecules are suggested as candidate in planta regulatory sites and as points for the artificial manipulation of GA-mediated growth control. CONCLUSIONS: This paper reviews: (1) the breakthrough dose-response experiments that show that root growth is regulated by GA in a lower concentration range than is required for shoot growth; (2) research on the regulation of GA biosynthesis pathways that are known predominantly to control shoot growth; and (3) recent research on GA signalling pathways, including GA receptors, which have been suggested to participate in GA-mediated growth regulation. This provides useful information to suggest a possible strategy for the selective control of shoot and root growth, and to explain how GA plays a role in rosette and liana plants with tall or short, and slender or thick axial organs.

• Mayerhofer H, Panneerselvam S, Mueller-Dieckmann J
• Protein kinase domain of CTR1 from Arabidopsis thaliana promotes ethylene receptor cross talk.
• J Mol Biol. 2012; 415: 768-79
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• Ethylene controls many aspects of plant growth and development. Signaling by the gaseous phytohormone is initiated by disulfide-linked membrane-bound receptors, and the formation of heteromeric receptor clusters contributes to the broad range of ethylene responsiveness. In Arabidopsis thaliana, the TCS-like ethylene receptors interact with the cytosolic serine/threonine kinase constitutive triple response 1 (CTR1), a proposed mitogen-activated protein kinase kinase kinase. In the absence of the hormone, the receptor and therefore CTR1 are active. Hence, ethylene acts as an inverse agonist of its signaling pathway. The three-dimensional structures of the active, triphosphorylated and the unphosphorylated, inactive kinase domain of CTR1 in complex with staurosporine illustrate the conformational rearrangements that form the basis of activity regulation. Additionally, in analytical ultracentrifugation experiments, active kinase domains form back-to-back dimers, while inactive and activation loop variants are monomers. Together with a front-to-front activation interface, the active protein kinase dimers thereby engage in interactions that promote CTR1-mediated cross talk between ethylene receptor clusters. This model provides a structural foundation for the observed high sensitivity of plants to ethylene.

• Band LR et al.
• Growth-induced hormone dilution can explain the dynamics of plant root cell elongation.
• Proc Natl Acad Sci U S A. 2012; 109: 7577-82
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• In the elongation zone of the Arabidopsis thaliana plant root, cells undergo rapid elongation, increasing their length by approximately 10-fold over 5 h while maintaining a constant radius. Although progress is being made in understanding how this growth is regulated, little consideration has been given as to how cell elongation affects the distribution of the key regulating hormones. Using a multiscale mathematical model and measurements of growth dynamics, we investigate the distribution of the hormone gibberellin in the root elongation zone. The model quantifies how rapid cell expansion causes gibberellin to dilute, creating a significant gradient in gibberellin levels. By incorporating the gibberellin signaling network, we simulate how gibberellin dilution affects the downstream components, including the growth-repressing DELLA proteins. We predict a gradient in DELLA that provides an explanation of the reduction in growth exhibited as cells move toward the end of the elongation zone. These results are validated at the molecular level by comparing predicted mRNA levels with transcriptomic data. To explore the dynamics further, we simulate perturbed systems in which gibberellin levels are reduced, considering both genetically modified and chemically treated roots. By modeling these cases, we predict how these perturbations affect gibberellin and DELLA levels and thereby provide insight into their altered growth dynamics.

• Westfall CS, Zubieta C, Herrmann J, Kapp U, Nanao MH, Jez JM
• Structural basis for prereceptor modulation of plant hormones by GH3 proteins.
• Science. 2012; 336: 1708-11
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• Acyl acid amido synthetases of the GH3 family act as critical prereceptor modulators of plant hormone action; however, the molecular basis for their hormone selectivity is unclear. Here, we report the crystal structures of benzoate-specific Arabidopsis thaliana AtGH3.12/PBS3 and jasmonic acid-specific AtGH3.11/JAR1. These structures, combined with biochemical analysis, define features for the conjugation of amino acids to diverse acyl acid substrates and highlight the importance of conformational changes in the carboxyl-terminal domain for catalysis. We also identify residues forming the acyl acid binding site across the GH3 family and residues critical for amino acid recognition. Our results demonstrate how a highly adaptable three-dimensional scaffold is used for the evolution of promiscuous activity across an enzyme family for modulation of plant signaling molecules.

• Hauvermale AL, Ariizumi T, Steber CM
• Gibberellin signaling: a theme and variations on DELLA repression.
• Plant Physiol. 2012; 160: 83-92

• Janssen BJ, Snowden KC
• Strigolactone and karrikin signal perception: receptors, enzymes, or both?
• Front Plant Sci. 2012; 3: 296-296
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• The signaling molecules strigolactone (SL) and karrikin are involved in seed germination, development of axillary meristems, senescence of leaves, and interactions with arbuscular mycorrhizal fungi. The signal transduction pathways for both SLs and karrikins require the same F-box protein (MAX2) and closely related alpha/beta hydrolase fold proteins (DAD2 and KAI2). The crystal structure of DAD2 has been solved revealing an alpha/beta hydrolase fold protein with an internal cavity capable of accommodating SLs. DAD2 responds to the SL analog GR24 by changing conformation and binding to MAX2 in a GR24 concentration-dependent manner. DAD2 can also catalyze hydrolysis of GR24. Structure activity relationships of analogs indicate that the butenolide ring common to both SLs and karrikins is essential for biological activity, but the remainder of the molecules can be significantly modified without loss of activity. The combination of data from the study of DAD2, KAI2, and chemical analogs of SLs and karrikins suggests a model for binding that requires nucleophilic attack by the active site serine of the hydrolase at the carbonyl atom of the butenolide ring. A conformational change occurs in the hydrolase that results in interaction with the F-box protein MAX2. Downstream signal transduction is then likely to occur via SCF (Skp-Cullin-F-box) complex-mediated ubiquitination of target proteins and their subsequent degradation. The role of the catalytic activity of the hydrolase is unclear but it may be integral in binding as well as possibly allowing the signal to be cleared from the receptor. The alpha/beta hydrolase fold family consists mostly of active enzymes, with a few notable exceptions. We suggest that DAD2 and KAI2 represent an intermediate stage where some catalytic activity is retained at the same time as a receptor role has evolved.

• Pearce S et al.
• Molecular characterization of Rht-1 dwarfing genes in hexaploid wheat.
• Plant Physiol. 2011; 157: 1820-31
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• The introduction of the Reduced height (Rht)-B1b and Rht-D1b semidwarfing genes led to impressive increases in wheat (Triticum aestivum) yields during the Green Revolution. The reduction in stem elongation in varieties containing these alleles is caused by a limited response to the phytohormone gibberellin (GA), resulting in improved resistance to stem lodging and yield benefits through an increase in grain number. Rht-B1 and Rht-D1 encode DELLA proteins, which act to repress GA-responsive growth, and their mutant alleles Rht-B1b and Rht-D1b are thought to confer dwarfism by producing more active forms of these growth repressors. While no semidwarfing alleles of Rht-A1 have been identified, we show that this gene is expressed at comparable levels to the other homeologs and represents a potential target for producing novel dwarfing alleles. In this study, we have characterized additional dwarfing mutations in Rht-B1 and Rht-D1. We show that the severe dwarfism conferred by Rht-B1c is caused by an intragenic insertion, which results in an in-frame 90-bp insertion in the transcript and a predicted 30-amino acid insertion within the highly conserved amino-terminal DELLA domain. In contrast, the extreme dwarfism of Rht-D1c is due to overexpression of the semidwarfing Rht-D1b allele, caused by an increase in gene copy number. We show also that the semidwarfing alleles Rht-B1d and Rht-B1e introduce premature stop codons within the amino-terminal coding region. Yeast two-hybrid assays indicate that these newly characterized mutations in Rht-B1 and Rht-D1 confer "GA-insensitive" dwarfism by producing DELLA proteins that do not bind the GA receptor GA INSENSITIVE DWARF1, potentially compromising their targeted degradation.

• Voegele A, Linkies A, Muller K, Leubner-Metzger G
• Members of the gibberellin receptor gene family GID1 (GIBBERELLIN INSENSITIVE DWARF1) play distinct roles during Lepidium sativum and Arabidopsis thaliana seed germination.
• J Exp Bot. 2011; 62: 5131-47
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• Germination of endospermic seeds is partly regulated by the micropylar endosperm, which acts as constraint to radicle protrusion. Gibberellin (GA) signalling pathways control coat-dormancy release, endosperm weakening, and organ expansion during seed germination. Three GIBBERELLIN INSENSITIVE DWARF1 (GID1) GA receptors are known in Arabidopsis thaliana: GID1a, GID1b, and GID1c. Molecular phylogenetic analysis of angiosperm GID1s reveals that they cluster into two eudicot (GID1ac, GID1b) groups and one monocot group. Eudicots have at least one gene from each of the two groups, indicating that the different GID1 receptors fulfil distinct roles during plant development. A comparative Brassicaceae approach was used, in which gid1 mutant and whole-seed transcript analyses in Arabidopsis were combined with seed-tissue-specific analyses of its close relative Lepidium sativum (garden cress), for which three GID1 orthologues were cloned. GA signalling via the GID1ac receptors is required for Arabidopsis seed germination, GID1b cannot compensate for the impaired germination of the gid1agid1c mutant. Transcript expression patterns differed temporarily, spatially, and hormonally, with GID1b being distinct from GID1ac in both species. Endosperm weakening is mediated, at least in part, through GA-induced genes encoding cell-wall-modifying proteins. A suppression subtraction hybridization (SSH) cDNA library enriched for sequences that are highly expressed during early germination in the micropylar endosperm contained expansins and xyloglucan endo-transglycosylases/hydrolases (XTHs). Their transcript expression patterns in both species strongly suggest that they are regulated by distinct GID1-mediated GA signalling pathways. The GID1ac and GID1b pathways seem to fulfil distinct regulatory roles during Brassicaceae seed germination and seem to control their downstream targets distinctly.

• Richter R, Behringer C, Muller IK, Schwechheimer C
• The GATA-type transcription factors GNC and GNL/CGA1 repress gibberellin signaling downstream from DELLA proteins and PHYTOCHROME-INTERACTING FACTORS.
• Genes Dev. 2010; 24: 2093-104
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• The phytohormone gibberellin (GA) regulates various developmental processes in plants such as germination, greening, elongation growth, and flowering time. DELLA proteins, which are degraded in response to GA, repress GA signaling by inhibitory interactions with PHYTOCHROME-INTERACTING FACTOR (PIF) family transcription factors. How GA signaling is controlled downstream from the DELLA and PIF regulators is, at present, unclear. Here, we characterize GNC (GATA, NITRATE-INDUCIBLE, CARBON-METABOLISM INVOLVED) and GNL/CGA1 (GNC-LIKE/CYTOKININ-RESPONSIVE GATA FACTOR1), two homologous GATA-type transcription factors from Arabidopsis thaliana that we initially identified as GA-regulated genes. Our genetic analyses of loss-of-function mutants and overexpression lines establish that GNC and GNL are functionally redundant regulators of germination, greening, elongation growth and flowering time. We further show by chromatin immunoprecipitation that both genes are potentially direct transcription targets of PIF transcription factors, and that their expression is up-regulated in pif mutant backgrounds. In line with a key role of GNC or GNL downstream from DELLA and PIF signaling, we find that their overexpression leads to gene expression changes that largely resemble those observed in a ga1 biosynthesis mutant or a pif quadruple mutant. These findings, together with the fact that gnc and gnl loss-of-function mutations suppress ga1 phenotypes, support the hypothesis that GNC and GNL are important repressors of GA signaling downstream from the DELLA and PIF regulators.

• Yamamoto Y et al.
• A rice gid1 suppressor mutant reveals that gibberellin is not always required for interaction between its receptor, GID1, and DELLA proteins.
• Plant Cell. 2010; 22: 3589-602
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• To investigate gibberellin (GA) signaling using the rice (Oryza sativa) GA receptor GIBBERELLIN-INSENSITIVE DWARF1 (GID1) mutant gid1-8, we isolated a suppressor mutant, Suppressor of gid1-1 (Sgd-1). Sgd-1 is an intragenic mutant containing the original gid1-8 mutation (L45F) and an additional amino acid substitution (P99S) in the loop region. GID1(P99S) interacts with the rice DELLA protein SLENDER RICE1 (SLR1), even in the absence of GA. Substitution of the 99th Pro with other amino acids revealed that substitution with Ala (P99A) caused the highest level of GA-independent interaction. Physicochemical analysis using surface plasmon resonance revealed that GID1(P99A) has smaller K(a) (association) and K(d) (dissociation) values for GA(4) than does wild-type GID1. This suggests that the GID1(P99A) lid is at least partially closed, resulting in both GA-independent and GA-hypersensitive interactions with SLR1. One of the three Arabidopsis thaliana GID1s, At GID1b, can also interact with DELLA proteins in the absence of GA, so we investigated whether GA-independent interaction of At GID1b depends on a mechanism similar to that of rice GID1(P99A). Substitution of the loop region or a few amino acids of At GID1b with those of At GID1a diminished its GA-independent interaction with GAI while maintaining the GA-dependent interaction. Soybean (Glycine max) and Brassica napus also have GID1s similar to At GID1b, indicating that these unique GID1s occur in various dicots and may have important functions in these plants.

• Yuan X, Yin P, Hao Q, Yan C, Wang J, Yan N
• Single amino acid alteration between valine and isoleucine determines the distinct pyrabactin selectivity by PYL1 and PYL2.
• J Biol Chem. 2010; 285: 28953-8
• Display abstract

• Abscisic acid (ABA) is one of the most important phytohormones in plant. PYL proteins were identified to be ABA receptors in Arabidopsis thaliana. Despite the remarkably high degree of sequence similarity, PYL1 and PYL2 exhibit distinct responses toward pyrabactin, an ABA agonist. PYL1 inhibits protein phosphatase type 2C upon binding of pyrabactin. In contrast, PYL2 appears relatively insensitive to this compound. The crystal structure of pyrabactin-bound PYL1 revealed that most of the PYL1 residues involved in pyrabactin binding are conserved, hence failing to explain the selectivity of pyrabactin for PYL1 over PYL2. To understand the molecular basis of pyrabactin selectivity, we determined the crystal structure of PYL2 in complex with pyrabactin at 1.64 A resolution. Structural comparison and biochemical analyses demonstrated that one single amino acid alteration between a corresponding valine and isoleucine determines the distinct pyrabactin selectivity by PYL1 and PYL2. These characterizations provide an important clue to dissecting the redundancy of PYL proteins.

• Romir J, Harter K, Stehle T
• Two-component systems in Arabidopsis thaliana--A structural view.
• Eur J Cell Biol. 2010; 89: 270-2
• Display abstract

• Plants respond to outside stimuli by initiating signaling cascades that regulate gene expression. Little structural information is available on the signaling proteins that are part of the two-component systems of plants, and how changes in phosphorylation translate into alterations of three-dimensional structures and changes in recognition domains remains largely mysterious. Our work is on deciphering aspects of these systems through the crystallization and structural analysis of two-component system proteins in Arabidopsis thaliana.

• Shibata N, Kagiyama M, Nakagawa M, Hirano Y, Hakoshima T
• Crystallization of the plant hormone receptors PYL9/RCAR1, PYL5/RCAR8 and PYR1/RCAR11 in the presence of (+)-abscisic acid.
• Acta Crystallogr Sect F Struct Biol Cryst Commun. 2010; 66: 456-9
• Display abstract

• Abscisic acid (ABA) is a plant hormone that plays key regulatory roles in physiological pathways for the adaptation of vegetative tissues to abiotic stresses such as water stress in addition to events pertaining to plant growth and development. The Arabidopsis ABA receptor proteins PYR/PYLs/RCARs form a START family that contains 14 members which are classified into three subfamilies (I-III). Here, purification, crystallization and X-ray data collection are reported for a member of each of the subfamilies, PYL9/RCAR1 from subfamily I, PYL5/RCAR8 from subfamily II and PYR1/RCAR11 from subfamily III, in the presence of (+)-abscisic acid. The three proteins crystallize in space groups P3(1)21/P3(2)21, P2 and P1, respectively. X-ray intensity data were collected to 1.9-2.6 A resolution.

• Santner A, Estelle M
• The ubiquitin-proteasome system regulates plant hormone signaling.
• Plant J. 2010; 61: 1029-40
• Display abstract

• Plants utilize the ubiquitin-proteasome system (UPS) to modulate nearly every aspect of growth and development. Ubiquitin is covalently attached to target proteins through the action of three enzymes known as E1, E2, and E3. The ultimate outcome of this post-translational modification depends on the nature of the ubiquitin linkage and the extent of polyubiquitination. In most cases, ubiquitination results in degradation of the target protein in the 26S proteasome. During the last 10 years it has become clear that the UPS plays a prominent regulatory role in hormone biology. E3 ubiquitin ligases in particular actively participate in hormone perception, de-repression of hormone signaling pathways, degradation of hormone specific transcription factors, and regulation of hormone biosynthesis. It is certain that additional functions will be discovered as more of the nearly 1200 potential E3s in plants are elucidated.

• Sun TP
• Gibberellin-GID1-DELLA: a pivotal regulatory module for plant growth and development.
• Plant Physiol. 2010; 154: 567-70

• Guo J, Wang S, Wang J, Huang WD, Liang J, Chen JG
• Dissection of the relationship between RACK1 and heterotrimeric G-proteins in Arabidopsis.
• Plant Cell Physiol. 2009; 50: 1681-94
• Display abstract

• Mammalian receptor for activated C kinase 1 (RACK1) is a versatile scaffold protein, playing regulatory roles in multiple signal transduction pathways. Moreover, RACK1 interacts with the heterotrimeric G-proteins (G-proteins) and regulates some specific functions of Gbetagamma. Although the protein sequences of both RACK1 and G-proteins are highly conserved in Arabidopsis, their relationship remains elusive. Here we provide genetic and biochemical evidence that Arabidopsis RACK1 and G-proteins may act through a mechanism that is distinct from their counterparts in mammals. Loss-of-function alleles of RACK1A (the most abundantly expressed RACK1 gene in Arabidopsis) do not appear to share morphological and developmental phenotypes with loss-of-function alleles of GPA1 (encoding the sole Galpha in Arabidopsis) or AGB1 (encoding the sole Gbeta in Arabidopsis). The analysis of gpa1 rack1a and agb1 rack1a double mutants suggested that the effect of RACK1A on morphological and developmental traits may occur independently of the presence or absence of the G-protein subunits. Although both RACK1A and G-protein subunits are negative regulators of ABA responses in the ABA inhibition of early seedling development, an additive ABA hypersensitivity was observed in gpa1 rack1a and agb1 rack1a double mutants. Biochemical analysis suggested that unlike their counterparts in mammals, RACK1 may not physically interact with AGB1. Taken together, these findings revealed some fundamental differences in the relationship of RACK1 and G-proteins between Arabidopsis and mammals.

• Koini MA et al.
• High temperature-mediated adaptations in plant architecture require the bHLH transcription factor PIF4.
• Curr Biol. 2009; 19: 408-13
• Display abstract

• Exposure of Arabidopsis plants to high temperature (28 degrees C) results in a dramatic change in plant development. Responses to high temperature include rapid extension of plant axes, leaf hyponasty, and early flowering. These phenotypes parallel plant responses to the threat of vegetational shade and have been shown to involve the hormone auxin. In this work, we demonstrate that high temperature-induced architectural adaptations are mediated through the bHLH transcriptional regulator PHYTOCHROME INTERACTING FACTOR 4 (PIF4). Roles for PIF4 have previously been established in both light and gibberellin (GA) signaling, through interactions with phytochromes and DELLA proteins, respectively. Mutants deficient in PIF4 do not display elongation responses or leaf hyponasty upon transfer to high temperature. High temperature-mediated induction of the auxin-responsive gene IAA29 is also abolished in these plants. An early flowering response to high temperature is maintained in pif4 mutants, suggesting that architectural and flowering responses operate via separate signaling pathways. The role of PIF4 in temperature signaling does not, however, appear to operate through interaction with either phytochrome or DELLA proteins, suggesting the existence of a novel regulatory mechanism. We conclude that PIF4 is an important component of plant high temperature signaling and integrates multiple environmental cues during plant development.

• Thomas C, Fricke I, Weyand M, Berken A
• 3D structure of a binary ROP-PRONE complex: the final intermediate for a complete set of molecular snapshots of the RopGEF reaction.
• Biol Chem. 2009; 390: 427-35
• Display abstract

• Guanine nucleotide exchange factors (GEFs) catalyze the activation of GTP-binding proteins (G proteins) in a multi-step reaction comprising intermediary complexes with and without nucleotide. Rho proteins of plants (ROPs) are activated by novel RopGEFs with a catalytic PRONE domain. We have previously characterized structures of GDP-bound ROP and a ternary complex between plant-specific ROP nucleotide exchanger (PRONE) and ROP including loosely bound GDP. Now, we complete the molecular snapshots of the RopGEF reaction with the nucleotide-free ROP-PRONE structure at 2.9 A. The binary complex surprisingly closely resembles the preceding ternary intermediate including an unusually intact P-loop in the G protein. A striking difference is the prominent contact of the invariant P-loop lysine to a conserved switch II glutamate in ROP, favoring a key role of this interaction in driving out the nucleotide. The nucleotide-free state is supported by additional interactions involving the essential WW-motif in PRONE. We propose that this GEF region stabilizes the intact P-loop conformation, which facilitates re-association with a new nucleotide and further promotes the overall exchange reaction. With our novel structure, we provide further insights into the nucleotide exchange mechanism and present a first example of the complete GEF reaction at a molecular level.

• Achard P et al.
• Gibberellin signaling controls cell proliferation rate in Arabidopsis.
• Curr Biol. 2009; 19: 1188-93
• Display abstract

• Plant growth involves the integration of many environmental and endogenous signals that together with the intrinsic genetic program determine plant size. At the cellular level, growth rate is regulated by the combined activity of two processes: cell proliferation and expansion. Gibberellins (GA) are plant-specific hormones that play a central role in the regulation of growth and development with respect to environmental variability. It is well established that GA promote growth through cell expansion by stimulating the destruction of growth-repressing DELLA proteins (DELLAs); however, their effects on cell proliferation remain unknown. Kinematic analysis of leaf and root meristem growth revealed a novel function of DELLAs in restraining cell production. Moreover, by visualizing the cell cycle marker cyclinB1::beta-glucuronidase in GA-signaling mutants, we show that GA modulate cell cycle activity in the root meristem via a DELLA-dependent mechanism. Accordingly, expressing gai (a nondegradable DELLA protein) solely in root meristem reduced substantially the number of dividing cells. We also show that DELLAs restrain cell production by enhancing the levels of the cell cycle inhibitors Kip-related protein 2 (KRP2) and SIAMESE (SIM). Therefore, DELLAs exert a general plant growth inhibitory activity by reducing both cell proliferation and expansion rates, enabling phenotypic plasticity.

• Hayashi K, Kamio S, Oono Y, Townsend LB, Nozaki H
• Toyocamycin specifically inhibits auxin signaling mediated by SCFTIR1 pathway.
• Phytochemistry. 2009; 70: 190-7
• Display abstract

• The auxins, plant hormones, play a crucial role in many aspects of plant development by regulating cell division, elongation and differentiation. Toyocamycin, a nucleoside-type antibiotic, was identified as auxin signaling inhibitor in a screen of microbial extracts for inhibition of the auxin-inducible reporter gene assay. Toyocamycin specifically inhibited auxin-responsive gene expression, but did not affect other hormone-inducible gene expression. Toyocamycin also blocked auxin-enhanced degradation of the Aux/IAA repressor modulated by the SCF(TIR1) ubiquitin-proteasome pathway without inhibiting proteolytic activity of proteasome. Furthermore, toyocamycin inhibited auxin-induced lateral root formation and epinastic growth of cotyledon in the Arabidopsis thaliana plant. This evidence suggested that toyocamycin would act on the ubiquitination process regulated by SCF(TIR1) machineries. To address the structural requirements for the specific activity of toyocamycin on auxin signaling, the structure-activity relationships of nine toyocamycin-related compounds, including sangivamycin and tubercidin, were investigated.

• Ohyama K, Shinohara H, Ogawa-Ohnishi M, Matsubayashi Y
• A glycopeptide regulating stem cell fate in Arabidopsis thaliana.
• Nat Chem Biol. 2009; 5: 578-80
• Display abstract

• The secreted peptide gene CLAVATA3 (CLV3) regulates stem cell fate in the shoot apical meristem in Arabidopsis thaliana plants, but the molecular structure of the active mature CLV3 peptide is controversial. Here, using nano-LC-MS/MS analysis of apoplastic peptides of A. thaliana plants overexpressing CLV3, we show that CLV3 is a 13-amino-acid arabinosylated glycopeptide. Post-translational arabinosylation of CLV3 is critical for its biological activity and high-affinity binding to its receptor CLV1.

• Nelson DC et al.
• Karrikins discovered in smoke trigger Arabidopsis seed germination by a mechanism requiring gibberellic acid synthesis and light.
• Plant Physiol. 2009; 149: 863-73
• Display abstract

• Discovery of the primary seed germination stimulant in smoke, 3-methyl-2H-furo[2,3-c]pyran-2-one (KAR1), has resulted in identification of a family of structurally related plant growth regulators, karrikins. KAR1 acts as a key germination trigger for many species from fire-prone, Mediterranean climates, but a molecular mechanism for this response remains unknown. We demonstrate that Arabidopsis (Arabidopsis thaliana), an ephemeral of the temperate northern hemisphere that has never, to our knowledge, been reported to be responsive to fire or smoke, rapidly and sensitively perceives karrikins. Thus, these signaling molecules may have greater significance among angiosperms than previously realized. Karrikins can trigger germination of primary dormant Arabidopsis seeds far more effectively than known phytohormones or the structurally related strigolactone GR-24. Natural variation and depth of seed dormancy affect the degree of KAR1 stimulation. Analysis of phytohormone mutant germination reveals suppression of KAR1 responses by abscisic acid and a requirement for gibberellin (GA) synthesis. The reduced germination of sleepy1 mutants is partially recovered by KAR1, which suggests that germination enhancement by karrikin is only partly DELLA dependent. While KAR1 has little effect on sensitivity to exogenous GA, it enhances expression of the GA biosynthetic genes GA3ox1 and GA3ox2 during seed imbibition. Neither abscisic acid nor GA levels in seed are appreciably affected by KAR1 treatment prior to radicle emergence, despite marked differences in germination outcome. KAR1 stimulation of Arabidopsis germination is light-dependent and reversible by far-red exposure, although limited induction of GA3ox1 still occurs in the dark. The observed requirements for light and GA biosynthesis provide the first insights into the karrikin mode of action.

• De Rybel B, Audenaert D, Beeckman T, Kepinski S
• The past, present, and future of chemical biology in auxin research.
• ACS Chem Biol. 2009; 4: 987-98
• Display abstract

• Research into the plant hormone auxin has always been tightly linked with the use of small molecules. In fact, most of the known players in auxin signaling and transport in the model plant Arabidopsis thaliana were identified by screening for resistance to auxin analogues. The use of high-throughput screening technologies has since yielded many novel molecules, opening the way for the identification of new target proteins to further elucidate known pathways. Here, we give an overview of well-established and novel molecules used in auxin research and highlight the current status and future perspectives of chemical biology approaches to auxin biology.

• Lorkovic ZJ
• Role of plant RNA-binding proteins in development, stress response and genome organization.
• Trends Plant Sci. 2009; 14: 229-36
• Display abstract

• RNA-binding proteins (RBPs) in eukaryotes have crucial roles in all aspects of post-transcriptional gene regulation. They are important governors of diverse developmental processes by modulating expression of specific transcripts. The Arabidopsis (Arabidopsis thaliana) genome encodes for more than 200 different RBPs, most of which are plant specific and are therefore likely to perform plant-specific functions. Indeed, recent identification and analysis of plant RBPs clearly showed that, in addition to the important role in diverse developmental processes, they are also involved in adaptation of plants to various environmental conditions. Clearly, they act by regulating pre-mRNA splicing, polyadenylation, RNA stability and RNA export, as well as by influencing chromatin modification.

• Hirano Y, Murase K, Hakoshima T
• [Structural studies of the plant hormone receptors].
• Tanpakushitsu Kakusan Koso. 2009; 54: 833-42

• Miyazono K et al.
• Structural basis of abscisic acid signalling.
• Nature. 2009; 462: 609-14
• Display abstract

• The phytohormone abscisic acid (ABA) mediates the adaptation of plants to environmental stresses such as drought and regulates developmental signals such as seed maturation. Within plants, the PYR/PYL/RCAR family of START proteins receives ABA to inhibit the phosphatase activity of the group-A protein phosphatases 2C (PP2Cs), which are major negative regulators in ABA signalling. Here we present the crystal structures of the ABA receptor PYL1 bound with (+)-ABA, and the complex formed by the further binding of (+)-ABA-bound PYL1 with the PP2C protein ABI1. PYL1 binds (+)-ABA using the START-protein-specific ligand-binding site, thereby forming a hydrophobic pocket on the surface of the closed lid. (+)-ABA-bound PYL1 tightly interacts with a PP2C domain of ABI1 by using the hydrophobic pocket to cover the active site of ABI1 like a plug. Our results reveal the structural basis of the mechanism of (+)-ABA-dependent inhibition of ABI1 by PYL1 in ABA signalling.

• Chini A, Fonseca S, Chico JM, Fernandez-Calvo P, Solano R
• The ZIM domain mediates homo- and heteromeric interactions between Arabidopsis JAZ proteins.
• Plant J. 2009; 59: 77-87
• Display abstract

• Discovery of the jasmonate ZIM-domain (JAZ) repressors defined the core jasmonate (JA) signalling module as COI1-JAZ-MYC2, and allowed a full view of the JA signalling pathway from hormone perception to transcriptional reprogramming. JAZ proteins are repressors of MYC2 and targets of SCF(COI1), which is the likely jasmonate receptor. Upon hormone perception, JAZ repressors are degraded by the proteasome releasing MYC2 and allowing the activation of JA responses. All members of the JAZ family share two conserved domains, the Jas motif, required for JAZ interactions with MYC2 and COI1, and the ZIM domain, the function of which is so far unknown. Here, we show that the ZIM domain acts as a protein-protein interaction domain mediating homo- and heteromeric interactions between JAZ proteins. These JAZ-JAZ interactions are independent of the presence of the hormone. The observation that only a few members of the JAZ family form homo- and heteromers may suggest the relevance of these proteins in the regulation of JA signalling. Interestingly, the JAZ3DeltaJas protein interacts with several JAZ proteins, providing new clues to understanding the dominant JA insensitivity promoted by truncated JAZDeltaJas proteins. We also provide evidence that the Jas motif mediates the hormone-dependent interaction between Arabidopsis JAZ3 and COI1, and further confirm that the Jas motif is required and sufficient for Arabidopsis JAZ3-MYC2 interaction. Finally, we show that interaction with MYC2 is a common feature of the JAZ family, as most JAZ proteins can bind MYC2 in pull-down and yeast two-hybrid assays.

• Backman HG, Pessoa J, Eneqvist T, Glaser E
• Binding of divalent cations is essential for the activity of the organellar peptidasome in Arabidopsis thaliana, AtPreP.
• FEBS Lett. 2009; 583: 2727-33
• Display abstract

• The dual-targeted mitochondrial and chloroplastic zinc metallooligopeptidase from Arabidopsis, AtPreP, functions as a peptidasome that degrades targeting peptides and other small unstructured peptides. In addition to Zn located in the catalytic site, AtPreP also contains two Mg-binding sites. We have investigated the role of Mg-binding using AtPreP variants, in which one or both sites were rendered unable to bind Mg(2+). Our results show that metal binding besides that of the active site is crucial for AtPreP proteolysis, particularly the inner site appears essential for normal proteolytic function. This is also supported by its evolutionary conservation among all plant species of PreP.

• Khan SA et al.
• A new strain of Arthrinium phaeospermum isolated from Carex kobomugi Ohwi is capable of gibberellin production.
• Biotechnol Lett. 2009; 31: 283-7
• Display abstract

• Plant growth-promoting endophytic fungi with gibberellin-producing ability were isolated from the roots of Carex kobomugi Ohwi, a common sand-dune plant, and bioassayed for plant growth-promotion. A new strain, Arthrinium phaeospermum KACC43901, promoted growth of waito-c rice and Atriplex gemelinii. Analysis of its culture filtrate showed the presence of bioactive GA(1) (0.5 ng/ml), GA(3) (8.8 ng/ml), GA(4) (4.7 ng/ml) and GA(7) (2.2 ng/ml) along with physiologically inactive GA(5) (0.4 ng/ml), GA(9) (0.6 ng/ml), GA(12) (0.4 ng/ml), GA(15) (0.4 ng/ml), GA(19) (0.9 ng/ml) and GA(24) (1.8 ng/ml). The fungal isolate was identified through sequence homology and phylogenetic analysis of 18S rDNA (internal transcribed region).

• Valdez HA, Busi MV, Wayllace NZ, Parisi G, Ugalde RA, Gomez-Casati DF
• Role of the N-terminal starch-binding domains in the kinetic properties of starch synthase III from Arabidopsis thaliana.
• Biochemistry. 2008; 47: 3026-32
• Display abstract

• Starch synthase III (SSIII), one of the SS isoforms involved in plant starch synthesis, has been reported to play a regulatory role in the synthesis of transient starch. SSIII from Arabidopsis thaliana contains 1025 amino acid residues and has an N-terminal transit peptide for chloroplast localization which is followed by three repeated starch-binding domains (SBDs; SSIII residues 22-591) and a C-terminal catalytic domain (residues 592-1025) similar to bacterial glycogen synthase. In this work, we constructed recombinant full-length and truncated isoforms of SSIII, lacking one, two, or three SBDs, and recombinant proteins, containing three, two, or one SBD, to investigate the role of these domains in enzyme activity. Results revealed that SSIII uses preferentially ADPGlc, although UDPGlc can also be used as a sugar donor substrate. When ADPGlc was used, the presence of the SBDs confers particular properties to each isoform, increasing the apparent affinity and the V max for the oligosaccharide acceptor substrate. However, no substantial changes in the kinetic parameters for glycogen were observed when UDPGlc was the donor substrate. Under glycogen saturating conditions, the presence of SBDs increases progressively the apparent affinity and V max for ADPGlc but not for UDPGlc. Adsorption assays showed that the N-terminal region of SSIII, containing three, two, or one SBD module have increased capacity to bind starch depending on the number of SBD modules, with the D23 protein (containing the second and third SBD module) being the one that makes the greatest contribution to binding. The results presented here suggest that the N-terminal SBDs have a regulatory role, showing a starch binding capacity and modulating the catalytic properties of SSIII.

• Pirrung MC et al.
• Ethylene receptor antagonists: strained alkenes are necessary but not sufficient.
• Chem Biol. 2008; 15: 313-21
• Display abstract

• Plants use ethylene as a hormone to control many physiological processes. Ethylene perception involves its binding to an unusual copper-containing, membrane-bound receptor. Inhibitors of ethylene action are valuable to study signaling and may have practical use in horticulture. Past investigation of alkene ligands for this receptor has identified strain as the key factor in antagonism of ethylene binding and action, consistent with known trends in metal-alkene complex stability. However, in this work, this principle could not be extended to other alkenes, prompting development of the proposal that a ring-opening reaction accounts for the unusual potency of cyclopropene ethylene antagonists. Another factor augmenting the affinity of alkenes for the copper binding site is pyramidalization, as in trans-cycloalkenes. The enantiomeric selectivity in the binding of one such alkene to the ethylene receptor demonstrates its protein-composed asymmetric environment.

• Symons GM, Smith JJ, Nomura T, Davies NW, Yokota T, Reid JB
• The hormonal regulation of de-etiolation.
• Planta. 2008; 227: 1115-25
• Display abstract

• De-etiolation involves a number of phenotypic changes as the plants shift from a dark-grown (etiolated) to a light-grown (de-etiolated) morphology. Whilst these light-induced, morphological changes are thought to be mediated by plant hormones, the precise mechanism/s are not yet fully understood. Here we provide further direct evidence that gibberellins (GAs) may play an important role in de-etiolation, because a similar light-induced reduction in bioactive GA levels was detected in barley (Hordeum vulgare L.), Arabidopsis (Arabidopsis thaliana L.), and pea (Pisum sativum L.). This is indicative of a highly conserved, negative-regulatory role for GAs in de-etiolation, in a range of taxonomically diverse species. In contrast, we found no direct evidence of a reduction in brassinosteroid (BR) levels during de-etiolation in any of these species.

• Aleman L et al.
• Functional analysis of cotton orthologs of GA signal transduction factors GID1 and SLR1.
• Plant Mol Biol. 2008; 68: 1-16
• Display abstract

• Gibberellic acid (GA) is both necessary and sufficient to promote fiber elongation in cultured fertilized ovules of the upland cotton variety Coker 312. This is likely due to the temporal and spatial regulation of GA biosynthesis, perception, and subsequent signal transduction that leads to alterations in gene expression and morphology. Our results indicate that the initiation of fiber elongation by the application of GA to cultured ovules corresponds with increased expression of genes that encode xyloglucan endotransglycosylase/hydrolase (XTH) and expansin (EXP) that are involved in promoting cell elongation. To gain a better understanding of the GA signaling components in cotton, that lead to such changes in gene expression, two GA receptor genes (GhGID1a and GhGID1b) and two DELLA protein genes (GhSLR1a and GhSLR1b) that are orthologous to the rice GA receptor (GID1) and the rice DELLA gene (SLR1), respectively, were characterized. Similar to the GA biosynthetic genes, expression of GhGID1a and GhGID1b is under the negative regulation by GA while GA positively regulates GhSLR1a. Recombinant GST-GhGID1s showed GA-binding activity in vitro that was augmented in the presence of GhSLR1a, GhSLR1b, or rice SLR1, indicating complex formation between the receptors and repressor proteins. This was further supported by the GA-dependent interaction of these proteins in yeast cells. Ectopic expression of the GhGID1a in the rice gid1-3 mutant plants rescued the GA-insensitive dwarf phenotype, which demonstrates that it is a functional GA receptor. Furthermore, ectopic expression of GhSLR1b in wild type Arabidopsis led to reduced growth and upregulated expression of DELLA-responsive genes.

• Achard P, Renou JP, Berthome R, Harberd NP, Genschik P
• Plant DELLAs restrain growth and promote survival of adversity by reducing the levels of reactive oxygen species.
• Curr Biol. 2008; 18: 656-60
• Display abstract

• Plant growth is adaptively modulated in response to environmental change. The phytohormone gibberellin (GA) promotes growth by stimulating destruction of the nuclear growth-repressing DELLA proteins [1-7], thus providing a mechanism for environmentally responsive growth regulation [8, 9]. Furthermore, DELLAs promote survival of adverse environments [8]. However, the relationship between these survival and growth-regulatory mechanisms was previously unknown. Here, we show that both mechanisms are dependent upon control of the accumulation of reactive oxygen species (ROS). ROS are small molecules generated during development and in response to stress that play diverse roles as eukaryotic intracellular second messengers [10]. We show that Arabidopsis DELLAs cause ROS levels to remain low after either biotic or abiotic stress, thus delaying cell death and promoting tolerance. In essence, stress-induced DELLA accumulation elevates the expression of genes encoding ROS-detoxification enzymes, thus reducing ROS levels. In accord with recent demonstrations that ROS control root cell expansion [11, 12], we also show that DELLAs regulate root-hair growth via a ROS-dependent mechanism. We therefore propose that environmental variability regulates DELLA activity [8] and that DELLAs in turn couple the downstream regulation of plant growth and stress tolerance through modulation of ROS levels.

• Yabe T et al.
• Structural analysis of Arabidopsis CnfU protein: an iron-sulfur cluster biosynthetic scaffold in chloroplasts.
• J Mol Biol. 2008; 381: 160-73
• Display abstract

• CnfU, a key iron-sulfur (Fe-S) cluster biosynthetic scaffold that is required for biogenesis of ferredoxin and photosystem I in chloroplasts, consists of two tandemly repeated domains in which only the N-terminal domain contains a conserved CXXC motif. We have determined the crystal structure of the metal-free dimer of AtCnfU-V from Arabidopsis thaliana at 1.35 A resolution. The N-terminal domains of the two monomers are linked together through two intermolecular disulfide bonds between the CXXC motifs. At the dimer interface, a total of four cysteine sulfur atoms provide a Fe-S cluster assembly site surrounded by uncharged but hydrophilic structurally mobile segments. The C-terminal domain of one monomer interacts with the N-terminal domain of the opposing monomer and thereby stabilizes dimer formation. Furthermore, Fe K-edge X-ray absorption spectroscopic analysis of the holo-CnfU dimer in solution suggests the presence of a typical [2Fe-2S]-type cluster coordinated by four thiolate ligands. Based on these data, a plausible model of the holo-AtCnfU-V dimer containing a surface-exposed [2Fe-2S] cluster assembled in the dimer interface was deduced. We propose that such a structural framework is important for CnfU to function as a Fe-S cluster biosynthetic scaffold.

• Ross JJ, Reid JB, Murfet IC, Weston DE
• The slender phenotype of pea is deficient in DELLA proteins.
• Plant Signal Behav. 2008; 3: 590-2
• Display abstract

• The recent cloning of the pea genes LA and CRY has historical implications, since the combined effect of null mutations in these genes is the elongated, gibberellin-insensitive "slender" phenotype, which gave rise to the theory that gibberellins (GAs) are inhibitors of inhibitors of growth. Interestingly, the duplication event that produced the second gene (LA or CRY) appears to have occurred more than 100 mya, and yet the two genes have retained essentially similar functions. They both encode DELLA proteins, which inhibit growth while at the same time promoting the synthesis of the growth-promoting hormone, gibberellin (GA). This duality of function is discussed in the context of recent suggestions that DELLAs integrate multiple hormone signals, rather than just the GA signal. We also present new data showing that LA and CRY play a major role in regulating fruit growth.

• Navarro L et al.
• DELLAs control plant immune responses by modulating the balance of jasmonic acid and salicylic acid signaling.
• Curr Biol. 2008; 18: 650-5
• Display abstract

• In Arabidopsis, the flagellin-derived peptide flg22 elevates antibacterial resistance [1] and inhibits growth [2] upon perception via the leucine-rich repeat receptor-like kinase Flagellin-Sensitive 2 (FLS2) [3]. DELLA proteins are plant growth repressors whose degradation is promoted by the phytohormone gibberellin [4]. Here, we show that DELLA stabilization contributes to flg22-induced growth inhibition. In addition, we show that DELLAs promote susceptibility to virulent biotrophs and resistance to necrotrophs, partly by altering the relative strength of salicylic acid and jasmonic acid (JA) signaling. A quadruple-DELLA mutant (which lacks four out of the five Arabidopsis DELLA proteins [5]) was partially insensitive to gene induction by Methyl-Jasmonate (MeJA), whereas the constitutively active dominant DELLA mutant gai[6] was sensitized for JA-responsive gene induction, implicating DELLAs in JA-signaling and/or perception. Accordingly, the elevated resistance of gai to the necrotrophic fungus Alternaria brassicicola and susceptibility to the hemibiotroph Pseudomonas syringae pv. tomato strain DC3000 (Pto DC3000) was attenuated in the JA-insensitive coi1-16 mutant [7]. These findings suggest an explanation for why the necrotrophic fungus Gibberella fujikuroi, causal agent of the foolish-seedling disease of rice, makes gibberellin.

• Kepinski S
• The anatomy of auxin perception.
• Bioessays. 2007; 29: 953-6
• Display abstract

• Auxin is a simple molecule but one with a complex and crucial influence on plant development. Accumulation and response to this important plant hormone underlies events as diverse as embryo patterning and growth responses to light and gravity. As such, research on auxin can be traced back to Darwin and has flourished into an immense body of work that has often had implications beyond plant biology. The latest instalment of the auxin story is no different:((1)) the solution of the crystal structure of the auxin receptor TIR1 illustrates, in spectacular detail, precisely how auxin is perceived and provides an insight into the working of a new class of receptor, which seems likely to be the first example of a new paradigm in eukaryotic signal transduction.

• Djakovic-Petrovic T, de Wit M, Voesenek LA, Pierik R
• DELLA protein function in growth responses to canopy signals.
• Plant J. 2007; 51: 117-26
• Display abstract

• Plants can sense neighbour competitors through light-quality signals and respond with shade-avoidance responses. These include increased shoot elongation, which enhances light capture and thus competitive power. Such plant-plant interactions therefore profoundly affect plant development in crowded populations. Shade-avoidance responses are tightly coordinated by interactions between light signals and hormones, with essential roles for the phytochrome B photoreceptor [sensing the red:far red (R:FR) ratio] and the hormone gibberellin (GA). The family of growth-suppressing DELLA proteins are targets for GA signalling and are proposed to integrate signals from other hormones. However, the importance of these regulators has not been studied in the ecologically relevant, complex realm of plant canopies. Here we show that DELLA abundance is regulated during growth responses to neighbours in dense Arabidopsis stands. This occurs in a R:FR-dependent manner in petioles, depends on GA, and matches the induction kinetics of petiole elongation. Similar interactions were observed in the growth response of seedling hypocotyls and are general for a second canopy signal, reduced blue light. Enhanced DELLA stability in the gai mutant inhibits shade-avoidance responses, indicating that DELLA proteins constrain shade-avoidance. However, using multiple DELLA knockout mutants, we show that the observed DELLA breakdown is not sufficient to induce shade-avoidance in petioles, but plays a more central role in hypocotyls. These data provide novel information on the regulation of shade-avoidance under ecologically important conditions, defining the importance of DELLA proteins and GA and unravelling the existence of GA- and DELLA-independent mechanisms.

• Vandenbussche F, Fierro AC, Wiedemann G, Reski R, Van Der Straeten D
• Evolutionary conservation of plant gibberellin signalling pathway components.
• BMC Plant Biol. 2007; 7: 65-65
• Display abstract

• BACKGROUND: Gibberellins (GA) are plant hormones that can regulate germination, elongation growth, and sex determination. They ubiquitously occur in seed plants. The discovery of gibberellin receptors, together with advances in understanding the function of key components of GA signalling in Arabidopsis and rice, reveal a fairly short GA signal transduction route. The pathway essentially consists of GID1 gibberellin receptors that interact with F-box proteins, which in turn regulate degradation of downstream DELLA proteins, suppressors of GA-controlled responses. RESULTS: Arabidopsis sequences of the gibberellin signalling compounds were used to screen databases from a variety of plants, including protists, for homologues, providing indications for the degree of conservation of the pathway. The pathway as such appears completely absent in protists, the moss Physcomitrella patens shares only a limited homology with the Arabidopsis proteins, thus lacking essential characteristics of the classical GA signalling pathway, while the lycophyte Selaginella moellendorffii contains a possible ortholog for each component. The occurrence of classical GA responses can as yet not be linked with the presence of homologues of the signalling pathway. Alignments and display in neighbour joining trees of the GA signalling components confirm the close relationship of gymnosperms, monocotyledonous and dicotyledonous plants, as suggested from previous studies. CONCLUSION: Homologues of the GA-signalling pathway were mainly found in vascular plants. The GA signalling system may have its evolutionary molecular onset in Physcomitrella patens, where GAs at higher concentrations affect gravitropism and elongation growth.

• Wojtas M et al.
• Cloning and characterization of Rab Escort Protein (REP) from Arabidopsis thaliana.
• Cell Biol Int. 2007; 31: 246-51
• Display abstract

• Intracellular vesicular trafficking is regulated by Rab proteins, small GTPases that require posttranslational geranylgeranylation for biological activity. This covalent modification is catalyzed by Rab geranylgeranyl transferase (RabGGTase) and proceeds only in the presence of accessory Rab Escort Protein (REP). In this communication, we report the cloning and characterization of REP gene of Arabidopsis thaliana. Highest expression of REP mRNA was detected in leaves and flowers in contrast to stems and roots. AtREP is recognized by anti-rat REP1 serum. Interaction of AtREP with the protein substrate is presented, as well as a structural model obtained through homology modeling, based on the known structure of rat REP1.

• Zentella R et al.
• Global analysis of della direct targets in early gibberellin signaling in Arabidopsis.
• Plant Cell. 2007; 19: 3037-57
• Display abstract

• Bioactive gibberellins (GAs) are phytohormones that regulate growth and development throughout the life cycle of plants. DELLA proteins are conserved growth repressors that modulate all aspects of GA responses. These GA-signaling repressors are nuclear localized and likely function as transcriptional regulators. Recent studies demonstrated that GA, upon binding to its receptor, derepresses its signaling pathway by binding directly to DELLA proteins and targeting them for rapid degradation via the ubiquitin-proteasome pathway. Therefore, elucidating the signaling events immediately downstream of DELLA is key to our understanding of how GA controls plant development. Two sets of microarray studies followed by quantitative RT-PCR analysis allowed us to identify 14 early GA-responsive genes that are also early DELLA-responsive in Arabidopsis thaliana seedlings. Chromatin immunoprecipitation provided evidence for in vivo association of DELLA with promoters of eight of these putative DELLA target genes. Expression of all 14 genes was downregulated by GA and upregulated by DELLA. Our study reveals that DELLA proteins play two important roles in GA signaling: (1) they help establish GA homeostasis by direct feedback regulation on the expression of GA biosynthetic and GA receptor genes, and (2) they promote the expression of downstream negative components that are putative transcription factors/regulators or ubiquitin E2/E3 enzymes. In addition, one of the putative DELLA targets, XERICO, promotes accumulation of abscisic acid (ABA) that antagonizes GA effects. Therefore, DELLA may restrict GA-promoted processes by modulating both GA and ABA pathways.

• Willige BC et al.
• The DELLA domain of GA INSENSITIVE mediates the interaction with the GA INSENSITIVE DWARF1A gibberellin receptor of Arabidopsis.
• Plant Cell. 2007; 19: 1209-20
• Display abstract

• Gibberellic acid (GA) promotes seed germination, elongation growth, and flowering time in plants. GA responses are repressed by DELLA proteins, which contain an N-terminal DELLA domain essential for GA-dependent proteasomal degradation of DELLA repressors. Mutations of or within the DELLA domain of DELLA repressors have been described for species including Arabidopsis thaliana, wheat (Triticum aestivum), maize (Zea mays), and barley (Hordeum vulgare), and we show that these mutations confer GA insensitivity when introduced into the Arabidopsis GA INSENSITIVE (GAI) DELLA repressor. We also demonstrate that Arabidopsis mutants lacking the three GA INSENSITIVE DWARF1 (GID1) GA receptor genes are GA insensitive with respect to GA-promoted growth responses, GA-promoted DELLA repressor degradation, and GA-regulated gene expression. Our genetic interaction studies indicate that GAI and its close homolog REPRESSOR OF ga1-3 are the major growth repressors in a GA receptor mutant background. We further demonstrate that the GA insensitivity of the GAI DELLA domain mutants is explained in all cases by the inability of the mutant proteins to interact with the GID1A GA receptor. Since we found that the GAI DELLA domain alone can mediate GA-dependent GID1A interactions, we propose that the DELLA domain functions as a receiver domain for activated GA receptors.

• Achard P et al.
• Integration of plant responses to environmentally activated phytohormonal signals.
• Science. 2006; 311: 91-4
• Display abstract

• Plants live in fixed locations and survive adversity by integrating growth responses to diverse environmental signals. Here, we show that the nuclear-localized growth-repressing DELLA proteins of Arabidopsis integrate responses to independent hormonal and environmental signals of adverse conditions. The growth restraint conferred by DELLA proteins is beneficial and promotes survival. We propose that DELLAs permit flexible and appropriate modulation of plant growth in response to changes in natural environments.

• Nakajima M et al.
• Identification and characterization of Arabidopsis gibberellin receptors.
• Plant J. 2006; 46: 880-9
• Display abstract

• Three gibberellin (GA) receptor genes (AtGID1a, AtGID1b and AtGID1c), each an ortholog of the rice GA receptor gene (OsGID1), were cloned from Arabidopsis, and the characteristics of their recombinant proteins were examined. The GA-binding activities of the three recombinant proteins were confirmed by an in vitro assay. Biochemical analyses revealed similar ligand selectivity among the recombinants, and all recombinants showed higher affinity to GA(4) than to other GAs. AtGID1b was unique in its binding affinity to GA(4) and in its pH dependence when compared with the other two, by only showing binding in a narrow pH range (pH 6.4-7.5) with 10-fold higher affinity (apparent K(d) for GA(4) = 3 x 10(-8) m) than AtGID1a and AtGID1c. A two-hybrid yeast system only showed in vivo interaction in the presence of GA(4) between each AtGID1 and the Arabidopsis DELLA proteins (AtDELLAs), negative regulators of GA signaling. For this interaction with AtDELLAs, AtGID1b required only one-tenth of the amount of GA(4) that was necessary for interaction between the other AtGID1s and AtDELLAs, reflecting its lower K(d) value. AtDELLA boosted the GA-binding activity of AtGID1 in vitro, which suggests the formation of a complex between AtDELLA and AtGID1-GA that binds AtGID1 to GA more tightly. The expression of each AtGID1 clone in the rice gid1-1 mutant rescued the GA-insensitive dwarf phenotype. These results demonstrate that all three AtGID1s functioned as GA receptors in Arabidopsis.

• Matsubayashi Y, Shinohara H, Ogawa M
• Identification and functional characterization of phytosulfokine receptor using a ligand-based approach.
• Chem Rec. 2006; 6: 356-64
• Display abstract

• Almost all plant cells, even when fully differentiated, can dedifferentiate and proliferate in vitro to form a callus, in which they can then differentiate to form various organs. These sequential processes can be promoted by exposing the cells to a conditioned medium in which either the same or other cells have previously been grown, indicating the involvement of cell-to-cell communication mediated by a chemical factor. This factor was purified from the conditioned medium and identified as a 5-amino-acid sulfated peptide. The addition of this peptide, named phytosulfokine (PSK), to the culture medium, even at nanomolar concentrations, significantly promotes cellular proliferation and/or cellular differentiation. We purified a membrane receptor for PSK (PSKR1) by ligand-based affinity chromatography and identified it as a member of leucine-rich repeat receptor kinases. The PSK-binding domain of PSKR1 was further identified by ligand photoaffinity labeling using a novel "on-column photoaffinity labeling" methodology that allows repeated incorporation of the photoaffinity label. Analysis of loss-of-function and gain-of-function mutants of the Arabidopsis PSKR1 revealed that PSK signaling affects cellular longevity and potential for growth without interfering with basic plant morphogenesis. These results suggest that PSK represents a new class of hormones that affect the potential for cellular growth and longevity of individual cells via binding to PSKR1, thereby exerting a pleiotropic effect on individual cells in response to environmental conditions.

• Itoh H et al.
• Dissection of the phosphorylation of rice DELLA protein, SLENDER RICE1.
• Plant Cell Physiol. 2005; 46: 1392-9
• Display abstract

• DELLA proteins are repressors of gibberellin signaling in plants. Our previous studies have indicated that gibberellin signaling is derepressed by SCF(GID2)-mediated proteolysis of the DELLA protein, SLENDER RICE1 (SLR1), in rice. In addition, the gibberellin-dependent increase of phosphorylated SLR1 in the loss-of-function gid2 mutant suggests that the SCF(GID2)-mediated degradation of SLR1 might be initiated by gibberellin-dependent phosphorylation. To confirm the role of phosphorylation of SLR1 in its gibberellin-dependent degradation, we revealed that SLR1 is phosphorylated on an N-terminal serine residue(s) within the DELLA/TVHYNP and polyS/T/V domain. However, gibberellin-induced phosphorylation in these regions was not observed in the gid2 mutant following the constitutive expression of SLR1 under the control of the rice actin1 promoter. Treatment with gibberellin induced both the phosphorylated and non-phosphorylated forms of SLR1 with similar induction kinetics in gid2 mutant cells. Both the phosphorylated and non-phosphorylated SLR1 proteins were degraded by gibberellin treatment with a similar half-life in the rice callus cells, and both proteins interacted with recombinant glutathione S-transferase (GST)-GID2. These results demonstrate that the phosphorylation of SLR1 is independent of its degradation and is dispensable for the interaction of SLR1 with the GID2/F-box protein.

• Fox BG et al.
• X-ray structure of Arabidopsis At1g77680, 12-oxophytodienoate reductase isoform 1.
• Proteins. 2005; 61: 206-8

• Rogers SW, Youn B, Rogers JC, Kang C
• Purification, crystallization and preliminary crystallographic studies of the ligand-binding domain of a plant vacuolar sorting receptor.
• Acta Crystallogr D Biol Crystallogr. 2004; 60: 2028-30
• Display abstract

• Vacuolar sorting receptor (VSR) proteins bind soluble protein ligands in a sequence-specific manner and target them to the lytic vacuole in plant cells. A VSR from Arabidopsis thaliana, AtBP80b, has been successfully purified after heterologous expression in Drosophila S2 cells. The AtBP80b protein (560 amino acids) was crystallized by the hanging-drop method with a PEG 400-based precipitant. Preliminary X-ray diffraction studies of an AtBP80b crystal showed that it belongs to the cubic space group P2(1)3 (or P4(2)32) and has unit-cell parameters a = b = c = 145.9 A. Crystals of the VSR diffract beyond 2.5 A resolution.

• Fu X, Richards DE, Fleck B, Xie D, Burton N, Harberd NP
• The Arabidopsis mutant sleepy1gar2-1 protein promotes plant growth by increasing the affinity of the SCFSLY1 E3 ubiquitin ligase for DELLA protein substrates.
• Plant Cell. 2004; 16: 1406-18
• Display abstract

• DELLA proteins restrain the cell proliferation and enlargement that characterizes the growth of plant organs. Gibberellin stimulates growth via 26S proteasome-dependent destruction of DELLAs, thus relieving DELLA-mediated growth restraint. Here, we show that the Arabidopsis thaliana sleepy1gar2-1 (sly1gar2-1) mutant allele encodes a mutant subunit (sly1gar2-1) of an SCF(SLY1) E3 ubiquitin ligase complex. SLY1 (the wild-type form) and sly1gar2-1 both confer substrate specificity on this complex via specific binding to the DELLA proteins. However, sly1gar2-1 interacts more strongly with the DELLA target than does SLY1. In addition, the strength of the SCFSLY1-DELLA interaction is increased by target phosphorylation. Growth-promoting DELLA destruction is dependent on SLY1 availability, on the strength of the interaction between SLY1 and the DELLA target, and on promotion of the SCFSLY1-DELLA interaction by DELLA phosphorylation.

• Spichal L et al.
• Two cytokinin receptors of Arabidopsis thaliana, CRE1/AHK4 and AHK3, differ in their ligand specificity in a bacterial assay.
• Plant Cell Physiol. 2004; 45: 1299-305
• Display abstract

• Strains of Escherichia coli that express two different cytokinin receptors of Arabidopsis thaliana, CRE1/AHK4 and AHK3, were used to study the relative sensitivity of these receptors to various cytokinins. Both receptors were most sensitive to the bases of the isoprenoid-type cytokinins trans-zeatin and isopentenyladenine but differed significantly in the recognition of other cytokinin compounds. In particular, CRE1/AHK4 recognized at 1 microm concentration only trans-zeatin while AHK3 recognized cis-zeatin and dihydrozeatin as well, although with a lower sensitivity. Similarly, CRE1/AHK4 was not activated by cytokinin ribosides and ribotides, but AHK3 was. Comparisons using the ARR5::GUS fusion gene as a cytokinin reporter in Arabidopsis showed similar relative degrees of responses in planta, except that cytokinins with aromatic side chains showed much higher activities than in the bacterial assay. These results indicate that the diverse cytokinin compounds might have specific functions in the numerous cytokinin-regulated processes, which may depend in turn on different receptors and their associated signalling pathways. The importance of precise control of local concentrations of defined cytokinin metabolites to regulate the respective downstream event is corroborated.

• Sugano M, Kaneyasu S, Nakamura T
• Gravitational and hormonal control in secondary xylem formation of Japanese flowering cherry.
• Biol Sci Space. 2003; 17: 245-6
• Display abstract

• It has been reported that Japanese flowering cherry promotes the formation of tension wood in the upper side of the inclined stem, which induces the negative gravitropism. In the other hand, the plant under simulated micro-gravity conditions reduces the width of the secondary xylem and increases the density of the vessels, which means decreased the density of the fiber cells. The plant under simulated micro-gravity conditions showed that the inhibition of xylem development and the decreased supporting mechanism in the stem. In this study we examined that the effects of auxin and gibberellin in the plant under simulated micro-gravity conditions and inclined stimulus. The gibberellin treatment promotes the secondary xylem development under simulated microgravity conditions and inclined stimulus. The upper side of the inclined stem contents much higher levels of IAA and gibberellin A1(GA1) than the lower side of that.

• Hirsch J, DeBeer George S, Solomon EI, Hedman B, Hodgson KO, Burstyn JN
• Raman and extended X-ray absorption fine structure characterization of a sulfur-ligated Cu(I) ethylene complex: modeling the proposed ethylene binding site of Arabidopsis thaliana ETR1.
• Inorg Chem. 2001; 40: 2439-41

• Fago G, Filippi A, Giardini A, Lagana A, Paladini A, Speranza M
• Chiral Recognition of O-Phosphoserine by Mass Spectrometry This work was supported by the Ministero della Universita e della Ricerca Scientifica e Tecnologica (MURST) and the Consiglio Nazionale delle Ricerche (CNR). The authors express their gratitude to F. Angelelli for technical assistance.
• Angew Chem Int Ed Engl. 2001; 40: 4051-4054

• Richards DE, King KE, Ait-Ali T, Harberd NP
• HOW GIBBERELLIN REGULATES PLANT GROWTH AND DEVELOPMENT: A Molecular Genetic Analysis of Gibberellin Signaling.
• Annu Rev Plant Physiol Plant Mol Biol. 2001; 52: 67-88
• Display abstract

• Gibberellins are hormones that control growth and a wide variety of other plant developmental processes. In recent years, significant progress has been made on the biochemistry of gibberellin biosynthesis and on the mechanisms by which gibberellin levels are regulated in plants. There have also been major advances in the understanding of gibberellin signaling, with several key genes being cloned. This review discusses our current understanding of gibberellin signaling, as seen from the perspective of molecular genetic analysis, and relates these observations to previous biochemical studies. In particular, we highlight an important conclusion of recent years: that GAI/RGA and orthologs play major roles in gibberellin signaling in diverse plant species, and that gibberellin probably stimulates growth by derepression of GAI/RGA.

• Hedden P
• Gibberellin Metabolism and Its Regulation.
• J Plant Growth Regul. 2001; 20: 317-318

• Ogas J
• Gibberellins
• Curr Biol. 2000; 10: 48-48

• Nakajima M et al.
• Partial purification and characterization of a gibberellin-binding protein from seedlings of Azukia angularis.
• Biochem Biophys Res Commun. 1997; 241: 782-6
• Display abstract

• Little is known about mechanisms for the perception of gibberellin (GA), a phytohormone, by plants. A GA-binding protein (GBP) with receptor-like characteristics was partially purified from azuki bean (Azukia angularis) seedlings. Gibberellins that caused much elongation of azuki epicotyl sections showed strong binding affinity for the GBP, and their biosynthetic precursors and inactive metabolites had weak affinity for it. The apparent Kd value for GA4 was calculated to be 7 x 10(-10) M from Scatchard plots. The molecular weight of the GBP in the native state was estimated to be 25,000 by high-performance liquid chromatography with a gel-permeation column. The GBP may be one of the receptor candidates for GAs.

• Swain SM, Olszewski NE
• Genetic Analysis of Gibberellin Signal Transduction.
• Plant Physiol. 1996; 112: 11-17

• Allan AC, Trewavas AJ
• Abscisic Acid and Gibberellin Perception: Inside or Out?
• Plant Physiol. 1994; 104: 1107-1108

• Jokhadze DI
• Gibberellin-binding proteins and hormonal regulation of transcription in cell nuclei and chloroplasts of higher plants.
• Subcell Biochem. 1991; 17: 327-40

• Atzorn R, Weiler EW
• The immunoassay of gibberellins : I. radioimmunoassays for the gibberellins A1, A 3, A 4, A 7, A 9 and A 20.
• Planta. 1983; 159: 1-6
• Display abstract

• Immunological assays for the detection of picogram amounts of gibberellins A1, A3, A4, A7, A9 and A20 are described. Antisera of high affinity (Ka over 10(10) l mol(-1)) were raised in rabbits by immunization against carboxyl-coupled gibberellin-bovine-serum-albumin conjugates and used in combination with tritiated gibberellins or gibberellin derivatives of high specific radioactivity (over 10(14) Bq mol(-1)). The assays allow gibberellin quantitation in as little as 1 mg of vegetative plant tissue.

• Sciuto S, Chillemi R, Piattelli M, Puglisi G
• Gibberellin- and cytokinin-like activities in marine algae from central Mediterranean.
• Boll Soc Ital Biol Sper. 1981; 57: 1590-5
• Display abstract

• The distribution of gibberellin- and cytokinin-like activities in twenty-one species of mediterranean algae belonging to Phaeophyta and Rhodophyta has been investigated. Fractions containing each of these activities have been obtained by partitioning aqueous-methanolic extracts of algae against ethyl acetate and butanol successively at different pH values. The stimulation or inhibition of amaranthin synthesis in Amaranthus sp. seedlings have been used a biological tests to assay the cytokinin- or gibberellin-like activities respectively. One or both these two hormonal activities have been detected in all species examined. While cytokinin-like activity has been found with the same frequency in the two phyla, the gibberellin-like activity was less frequent in Phaeophyta.

• Coombe BG
• GA32: A Polar Gibberellin with High Biological Potency.
• Science. 1971; 172: 856-7
• Display abstract

• A new gibberellin, GA(32), purified from Prunus seed, has three interesting properties: (i) it is the most polar of the 34 gibberellins presently known, (ii) it is the most potent of all gibberellins tested in the barley endosperm bioassay, and (iii) it is very active in promoting the growth of unpollinated apricot ovaries with a normal ratio of length to diameter.

• Moblidowska I
• Effects of some growth regulatiors on frost damage.
• Cryobiology. 1968; 5: 175-87

• Varga M, Bito M
• On the mechanism of gibberellin--auxin interaction. I. Effect of gibberellin on the quantity of free IAA and IAA conjugates in bean hypocotyl tissues.
• Acta Biol Acad Sci Hung. 1968; 19: 445-53

• HANSON JR, MULHOLLAND TP
• GIBBERELLIC ACID. XXIX. SOME TRANSFORMATION OF GIBBERELLIN A9.
• J Chem Soc. 1965; 33: 3550-5

• PODOJIL M, RICICOVA A
• INFLUENCE OF THE SOYA MEAL FRACTIONS ON GIBBERELLIC ACID AND GIBBERELLIN A PRODUCTION IN SUBMERSE CULTIVATION OF GIBBERELLA FUJIKUROI.
• Folia Microbiol (Praha). 1965; 10: 55-9

• Voeller BR
• Gibberellins: Their Effect on Antheridium Formation in Fern Gametophytes.
• Science. 1964; 143: 373-5
• Display abstract

• Gametophytes of the fern Anemia phyllitidis respond to seven different gibberellins (A(1), A(3); A(4), A(5), A(7), A(8), and A(9)) by forming antheridia. Gibberellin A(7) is active at concentrations as low as 5 X 10(-10) g/ ml. Species from two of the three other genera in the family Schizaeaceae respond similarly to gibberellin A(3). In contrast, nearly 40 species from 7 other families of ferns do not develop antheridia when supplied with gibberellin.

• MOWAT JA
• GIBBERELLIN-LIKE SUBSTANCES IN ALGAE.
• Nature. 1963; 200: 453-5

• BAUMGARTNER WE, LAZER L, DALZIEL A
• Application of tritium in the determination of gibberellins.
• Adv Tracer Methodol. 1963; 1: 257-62

• KATO J, PURVES WK, PHINNEY BO
• Gibberellin-like substances in plants.
• Nature. 1962; 196: 687-8

• HABER AH, CARRIER WL, ENOCHS NJ
• Gibberellin action on growth of seedings in the absence of photosynthesis.
• Nature. 1961; 190: 1034-5

• STRAUS J, EPP RR
• Response of Cupressus funebris tissue cultures to gibberellins.
• Science. 1960; 131: 1806-7
• Display abstract

• Gibberellins are a specific growth requirement of tissue cultures derived from the staminate cones of Cupressus funebris. The tissue originally required coconut milk for growth. However, a mixture of acid-hydrolyzed casein hydrolyzate and gibberellins replaces the coconut milk entirely and permits even better growth. It is tentatively suggested that coconut milk contains gibberellins or related substances and that these substances are involved in the nitrogen metabolism of the tissue.

• Wittwer SH, Tolbert NE
• 2-Chloroethyl Trimethylammonium Chloride and Related Compounds as Plant Growth Substances. V. Growth, Flowering, and Fruiting Responses as Related to Those Induced by Auxin and Gibberellin.
• Plant Physiol. 1960; 35: 871-7

• DANCER J
• Synergistic effect of zinc and gibberellin.
• Nature. 1959; 183: 901-2

• MARGARA J
• [Comparison of the action of gibberellic acid on the genus Beta].
• C R Hebd Seances Acad Sci. 1959; 249: 751-3

• BENVENUTO R
• [La circolazione extracorporea; tecnica della perfusione].
• Minerva Med. 1958; 49: 4111-4

• Sachs RM, Lang A
• Effect of Gibberellin on Cell Division in Hyoscyamus.
• Science. 1957; 125: 1144-5

• SILVESTRONI E, BIANCO I
• Ricerche sulla frequenza della microcitemia in Sicilia.
• Policlinico Prat. 1949; 56: 602-4

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