RESUMO
The formation of leaves and other lateral organs in plants depends on the proper specification of adaxial-abaxial (upper-lower) polarity. KANADI1 (KAN1), a member of the GARP family of transcription factors, is a key regulator of abaxial identity, leaf growth, and meristem formation in Arabidopsis thaliana. Here, we demonstrate that the Myb-like domain in KAN1 binds the 6-bp motif GNATA(A/T) and that this motif alone is sufficient to squelch transcription of a linked reporter in vivo. In addition, we report that KAN1 acts as a transcriptional repressor. Among its targets are genes involved in auxin biosynthesis, auxin transport, and auxin response. Furthermore, we find that the adaxializing HD-ZIPIII transcription factor REVOLUTA has opposing effects on multiple components of the auxin pathway. We hypothesize that HD-ZIPIII and KANADI transcription factors pattern auxin accumulation and responsiveness in the embryo. Specifically, we propose the opposing actions of KANADI and HD-ZIPIII factors on cotyledon formation (KANADI represses and HD-ZIPIII promotes cotyledon formation) occur through their opposing actions on genes acting at multiple steps in the auxin pathway.
Assuntos
Proteínas de Arabidopsis/fisiologia , Arabidopsis/metabolismo , Proteínas de Homeodomínio/fisiologia , Ácidos Indolacéticos/metabolismo , Proteínas Repressoras/fisiologia , Proteínas de Arabidopsis/metabolismo , Transporte Biológico , Regulação da Expressão Gênica de Plantas , Redes Reguladoras de Genes , Proteínas de Homeodomínio/metabolismo , Proteínas Repressoras/genética , Proteínas Repressoras/metabolismo , Transdução de Sinais/genética , Fatores de TranscriçãoRESUMO
The broadly conserved Class III homeodomain leucine zipper (HD-ZIPIII) and KANADI transcription factors have opposing and transformational effects on polarity and growth in all tissues and stages of the plant's life. To obtain a comprehensive understanding of how these factors work, we have identified transcripts that change in response to induced HD-ZIPIII or KANADI function. Additional criteria used to identify high-confidence targets among this set were presence of an adjacent HD-ZIPIII binding site, expression enriched within a subdomain of the shoot apical meristem, mutant phenotype showing defect in polar leaf and/or meristem development, physical interaction between target gene product and HD-ZIPIII protein, opposite regulation by HD-ZIPIII and KANADI, and evolutionary conservation of the regulator-target relationship. We find that HD-ZIPIII and KANADI regulate tissue-specific transcription factors involved in subsidiary developmental decisions, nearly all major hormone pathways, and new actors (such as indeterminate domain4) in the ad/abaxial regulatory network. Multiple feedback loops regulating HD-ZIPIII and KANADI are identified, as are mechanisms through which HD-ZIPIII and KANADI oppose each other. This work lays the foundation needed to understand the components, structure, and workings of the ad/abaxial regulatory network directing basic plant growth and development.
Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/genética , Regulação da Expressão Gênica de Plantas , Redes Reguladoras de Genes/genética , Proteínas de Homeodomínio/genética , Fatores de Transcrição/genética , Arabidopsis/anatomia & histologia , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Sítios de Ligação , Padronização Corporal , Regulação para Baixo , Expressão Gênica , Perfilação da Expressão Gênica , Proteínas de Homeodomínio/metabolismo , Meristema/anatomia & histologia , Meristema/genética , Meristema/crescimento & desenvolvimento , Meristema/metabolismo , Modelos Biológicos , Mutação , Análise de Sequência com Séries de Oligonucleotídeos , Fenótipo , Filogenia , Folhas de Planta/anatomia & histologia , Folhas de Planta/genética , Folhas de Planta/crescimento & desenvolvimento , Folhas de Planta/metabolismo , Brotos de Planta/anatomia & histologia , Brotos de Planta/genética , Brotos de Planta/crescimento & desenvolvimento , Brotos de Planta/metabolismo , Plantas Geneticamente Modificadas , Fatores de Transcrição/metabolismo , Regulação para CimaRESUMO
Spatiotemporal control of the formation of organ primordia and organ boundaries from the stem cell niche in the shoot apical meristem (SAM) determines the patterning and architecture of plants, but the underlying signaling mechanisms remain poorly understood. Here we show that brassinosteroids (BRs) play a key role in organ boundary formation by repressing organ boundary identity genes. BR-hypersensitive mutants display organ-fusion phenotypes, whereas BR-insensitive mutants show enhanced organ boundaries. The BR-activated transcription factor BZR1 directly represses the cup-shaped cotyledon (CUC) family of organ boundary identity genes. In WT plants, BZR1 accumulates at high levels in the nuclei of central meristem and organ primordia but at a low level in organ boundary cells to allow CUC gene expression. Activation of BR signaling represses CUC gene expression and causes organ fusion phenotypes. This study uncovers a role for BR in the spatiotemporal control of organ boundary formation and morphogenesis in the SAM.
Assuntos
Arabidopsis/genética , Brassinosteroides/metabolismo , Regulação da Expressão Gênica de Plantas , Meristema/fisiologia , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Núcleo Celular/metabolismo , Imunoprecipitação da Cromatina , Genes de Plantas , Microscopia Confocal/métodos , Microscopia Eletrônica de Varredura/métodos , Mutação , Fenótipo , Brotos de Planta/metabolismo , Transdução de SinaisRESUMO
Class III homeodomain leucine zipper (HD-ZIP III) transcription factors regulate critical developmental programs in plants; these include leaf polarity, polarity along the shoot-root axis, and stem cell specification and proliferation. One of the defining features of HD-ZIP III proteins is the presence of a Per-ARNT-Sim-like (PAS-like) MEKHLA domain at the C terminus. PAS-like domains are known to respond to a variety of chemical and physical stimuli. Here, we provide evidence that the MEKHLA domain acts as a negative regulator of Arabidopsis thaliana HD-ZIP III REVOLUTA activity. Based on experiments in yeast and plants, we propose a model in which the MEKHLA domain inhibits dimerization through a sequence-independent steric masking mechanism. This inhibition is relieved in response to a cellular signal that requires the C terminus of the MEKHLA domain for its perception. Overexpression experiments suggest that this signal is unequally distributed and/or sensed in the plant. Our data show that the function of the REVOLUTA MEKHLA domain differs among other HD-ZIP III family members; this difference may explain the genetic differences that have been observed among family members. This finding, combined with our phylogenetic analysis, suggests that REVOLUTA is the latest type of HD-ZIP III protein to have evolved in land plants.
Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/genética , Proteínas de Homeodomínio/metabolismo , Domínios e Motivos de Interação entre Proteínas , Sequência de Aminoácidos , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Clonagem Molecular , Biologia Computacional , Proteínas de Homeodomínio/genética , Zíper de Leucina , Dados de Sequência Molecular , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Multimerização Proteica , Alinhamento de SequênciaRESUMO
Unlike the situation in animals, the final morphology of the plant body is highly modulated by the environment. During Arabidopsis development, intrinsic factors provide the framework for basic patterning processes. CLASS III HOMEODOMAIN LEUCINE ZIPPER (HD-ZIPIII) transcription factors are involved in embryo, shoot and root patterning. During vegetative growth HD-ZIPIII proteins control several polarity set-up processes such as in leaves and the vascular system. We have identified several direct target genes of the HD-ZIPIII transcription factor REVOLUTA (REV) using a chromatin immunoprecipitation/DNA sequencing (ChIP-Seq) approach. This analysis revealed that REV acts upstream of auxin biosynthesis and affects directly the expression of several class II HD-ZIP transcription factors that have been shown to act in the shade-avoidance response pathway. We show that, as well as involvement in basic patterning, HD-ZIPIII transcription factors have a critical role in the control of the elongation growth that is induced when plants experience shade. Leaf polarity is established by the opposed actions of HD-ZIPIII and KANADI transcription factors. Finally, our study reveals that the module that consists of HD-ZIPIII/KANADI transcription factors controls shade growth antagonistically and that this antagonism is manifested in the opposed regulation of shared target genes.
Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/genética , Padronização Corporal , Proteínas de Homeodomínio/genética , Fatores de Transcrição/genética , Adaptação Fisiológica , Arabidopsis/citologia , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/efeitos da radiação , Proteínas de Arabidopsis/metabolismo , Sítios de Ligação , Imunoprecipitação da Cromatina , Regulação da Expressão Gênica no Desenvolvimento , Regulação da Expressão Gênica de Plantas , Genoma de Planta/genética , Proteínas de Homeodomínio/metabolismo , Hipocótilo/citologia , Hipocótilo/genética , Hipocótilo/crescimento & desenvolvimento , Hipocótilo/efeitos da radiação , Hibridização In Situ , Ácidos Indolacéticos/análise , Ácidos Indolacéticos/metabolismo , Luz , Mutação , Filogenia , Folhas de Planta/citologia , Folhas de Planta/genética , Folhas de Planta/crescimento & desenvolvimento , Folhas de Planta/efeitos da radiação , Análise de Sequência de DNA , Transdução de Sinais , Fatores de Transcrição/metabolismoRESUMO
In this issue of Developmental Cell, Nodine and colleagues show that two related leucine-rich repeat receptor kinases, RECEPTOR-LIKE PROTEIN KINASE1 and TOADSTOOL2, are critical in establishing radial pattern in the Arabidopsis embryo (Nodine et al., 2007). Embryos lacking these kinases show replacement of outer cell fates with inner cell fates.
Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/embriologia , Desenvolvimento Embrionário , Regulação da Expressão Gênica de Plantas , Proteínas Quinases/metabolismo , Proteínas de Arabidopsis/genética , Proteínas Quinases/genéticaAssuntos
Arabidopsis , Etilenos/metabolismo , Células-Tronco , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Divisão Celular , Proteínas Quinases/genética , Proteínas Quinases/metabolismo , Células-Tronco/citologia , Células-Tronco/metabolismoRESUMO
Dominant mutations in the Arabidopsis PHABULOSA (PHB) and PHAVOLUTA (PHV) transcription factor genes cause transformation of abaxial to adaxial leaf fates by altering a microRNA complementary site present in processed PHB and PHV mRNAs but not in the corresponding genomic DNA. phb-1d mutants accumulate excess PHB transcript throughout the leaf primordium, indicating defective regulation of PHB transcript synthesis and/or stability. We show that PHB and PHV coding sequences are heavily methylated downstream of the microRNA complementary site in most wild-type plant cells and that methylation is reduced in phb-1d and phv-1d mutants. Decreased methylation is limited to the chromosome bearing the dominant mutant allele in phb-1d heterozygotes. Low levels of methylation are detected in wt PHB DNA isolated from undifferentiated tissues. These results suggest a model in which the microRNA interacts with nascent, newly processed PHB mRNA to alter chromatin of the corresponding PHB template DNA predominantly in differentiated cells.
Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Cromossomos de Plantas , Metilação de DNA , MicroRNAs/metabolismo , RNA Mensageiro/metabolismo , Alelos , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Proteínas de Arabidopsis/química , Proteínas de Arabidopsis/genética , Sequência de Bases , Sítios de Ligação , Mapeamento Cromossômico , Ilhas de CpG , DNA de Plantas/efeitos dos fármacos , Éxons , Genes Dominantes , Genes de Plantas , Heterozigoto , MicroRNAs/química , Modelos Biológicos , Dados de Sequência Molecular , Mutação , RNA Mensageiro/classificação , RNA de Plantas/química , RNA de Plantas/isolamento & purificação , RNA de Plantas/metabolismo , Homologia de Sequência do Ácido Nucleico , Sulfitos/toxicidade , Moldes Genéticos , Fatores de Transcrição/genéticaRESUMO
Drought inhibits plant growth and can also induce premature senescence. Here we identify a transcription factor, ABA INSENSITIVE GROWTH 1 (ABIG1) required for abscisic acid (ABA) mediated growth inhibition, but not for stomatal closure. ABIG1 mRNA levels are increased both in response to drought and in response to ABA treatment. When treated with ABA, abig1 mutants remain greener and produce more leaves than comparable wild-type plants. When challenged with drought, abig1 mutants have fewer yellow, senesced leaves than wild-type. Induction of ABIG1 transcription mimics ABA treatment and regulates a set of genes implicated in stress responses. We propose a model in which drought acts through ABA to increase ABIG1 transcription which in turn restricts new shoot growth and promotes leaf senescence. The results have implications for plant breeding: the existence of a mutant that is both ABA resistant and drought resistant points to new strategies for isolating drought resistant genetic varieties.
Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/fisiologia , Secas , Regulação da Expressão Gênica de Plantas , Desenvolvimento Vegetal , Estresse Fisiológico , Fatores de Transcrição/metabolismo , Transcrição Gênica , Ácido Abscísico/metabolismo , Reguladores de Crescimento de Plantas/metabolismoRESUMO
The apical domain of the embryo is partitioned into distinct regions that will give rise to the cotyledons and the shoot apical meristem. In this article, we describe a novel screen to identify Arabidopsis thaliana embryo arrest mutants that are defective in this partitioning, and we describe the phenotype of one such mutant, bobber1. bobber1 mutants arrest at the globular stage of development, they express the meristem-specific SHOOTMERISTEMLESS gene throughout the top half of the embryo, and they fail to express the AINTEGUMENTA transcript normally found in cotyledons. Thus, BOBBER1 is required to limit the extent of the meristem domain and/or to promote the development of the cotyledon domains. Based on expression of early markers for apical development, bobber1 mutants differentiate protodermis and undergo normal early apical development. Consistent with a role for auxin in cotyledon development, BOBBER1 mutants fail to express localized maxima of the DR5:green fluorescent protein reporter. BOBBER1 encodes a protein with homology to the Aspergillus nidulans protein NUDC that has similarity to protein chaperones, indicating a possible role for BOBBER1 in synthesis or transport of proteins involved in patterning the Arabidopsis embryo.
Assuntos
Proteínas de Arabidopsis/fisiologia , Arabidopsis/embriologia , Sementes/embriologia , Sequência de Aminoácidos , Arabidopsis/genética , Proteínas de Arabidopsis/química , Proteínas de Arabidopsis/genética , Proteínas Fúngicas/química , Proteínas Fúngicas/genética , Regulação da Expressão Gênica de Plantas/genética , Regulação da Expressão Gênica de Plantas/fisiologia , Hibridização In Situ , Microscopia Confocal , Dados de Sequência Molecular , Plantas Geneticamente Modificadas/embriologia , Plantas Geneticamente Modificadas/genética , Estrutura Terciária de Proteína/genética , Estrutura Terciária de Proteína/fisiologia , Sementes/genética , Análise de Sequência de DNA , Homologia de Sequência de AminoácidosRESUMO
For nearly a century, the plant hormone auxin has been recognized for its effects on post-embryonic plant growth. Now recent insights into the molecular mechanism of auxin transport and signaling are uncovering fundamental roles for auxin in the earliest stages of plant development, such as in the development of the apical-basal (shoot-root) axis in the embryo, as well as in the formation of the root and shoot apical meristems and the cotyledons. Localized surges in auxin within the embryo occur through a sophisticated transcellular transport pathway causing the proteolytic destruction of key transcriptional repressors. As we discuss here, the resulting downstream gene activation, together with other, less well-understood regulatory pathways, establish much of the basic body plan of the angiosperm embryo.
Assuntos
Arabidopsis/embriologia , Desenvolvimento Embrionário/fisiologia , Ácidos Indolacéticos/metabolismo , Arabidopsis/anatomia & histologia , Arabidopsis/fisiologia , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/metabolismo , Ativação TranscricionalRESUMO
Pattern formation and morphogenesis require coordination of cell division rates and orientations with developmental signals that specify cell fate. A viable mutation in the TILTED1 locus, which encodes the catalytic subunit of DNA polymerase epsilon of Arabidopsis thaliana, causes a lengthening of the cell cycle by approximately 35% throughout embryo development and alters cell type patterning of the hypophyseal lineage in the root, leading to a displacement of the root pole from its normal position on top of the suspensor. Treatment of preglobular and early globular stages, but not later stage, embryos with the DNA polymerase inhibitor aphidicolin leads to a similar phenotype. The results uncover an interaction between the cell cycle and the processes that determine cell fate during plant embryogenesis.
Assuntos
Arabidopsis/embriologia , Padronização Corporal , Ciclo Celular , DNA Polimerase II/genética , Mutação , Sementes/crescimento & desenvolvimento , Sequência de Aminoácidos , Arabidopsis/citologia , Arabidopsis/enzimologia , Sequência de Bases , Catálise , Clonagem Molecular , DNA Polimerase II/química , Primers do DNA , DNA de Plantas/metabolismo , Dados de Sequência Molecular , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Sementes/metabolismo , Homologia de Sequência de AminoácidosRESUMO
Recent progress has been made in the genetic dissection of angiosperm shoot apical meristem (SAM) structure and function. Genes required for proper SAM development have been identified in a variety of species through the isolation of mutants. In addition, genes with expression patterns indicating they play a role in SAM function have been identified molecularly. The processes of SAM formation, self-renewal, and pattern formation within the SAM are examined with an emphasis on the contributions of recent classical and molecular genetic experiments to our understanding of this basic problem in plant developmental biology.
RESUMO
Plants produce proximal-distal growth axes with two types of growth potential: they can be indeterminate, in which case growth continues indefinitely, or they can be determinate, in which case growth is limited to the production of a single organ or a discrete set of organs. The indeterminate shoot axes of Arabidopsis pinhead/zwille mutants frequently are transformed to a determinate state. PINHEAD (PNH) is expressed in the central domain of the developing plant: the provascular tissue, the shoot apical meristem, and the adaxial (upper) sides of lateral organ primordia. Here, we show that ectopic expression of PNH on the abaxial (lower) sides of lateral organs results in upward curling of leaf blades. This phenotype correlates with a loss of cell number coordination between the two surfaces of the blade, indicating that ectopic PNH can cause changes in cell division rates. More strikingly, moving PNH expression from the central to the peripheral domain of the embryo causes transformation of the determinate cotyledon axis to an indeterminate state. We propose that growth axes are specified as determinate versus indeterminate in a PNH-mediated step. Our results add to a growing body of evidence that radial positional information is important in meristem formation. These results also indicate that genes regulating cell division and axis determinacy are likely to be among PNH targets.
Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/ultraestrutura , Proteínas de Arabidopsis/metabolismo , Cotilédone/genética , Cotilédone/crescimento & desenvolvimento , Cotilédone/ultraestrutura , Regulação da Expressão Gênica no Desenvolvimento , Regulação da Expressão Gênica de Plantas , Meristema/genética , Meristema/crescimento & desenvolvimento , Microscopia Eletrônica de Varredura , Mutação , Fenótipo , Plantas Geneticamente Modificadas , Sementes/genética , Sementes/crescimento & desenvolvimentoRESUMO
We describe a novel phenotype in Arabidopsis embryos homozygous for the temperature-sensitive topless-1 mutation. This mutation causes the transformation of the shoot pole into a root. Developing topless embryos fail to express markers for the shoot apical meristem (SHOOT MERISTEMLESS and UNUSUAL FLORAL ORGANS) and the hypocotyl (KNAT1). By contrast, the pattern of expression of root markers is either duplicated (LENNY, J1092) or expanded (SCARECROW). Shifts of developing topless embryos between permissive and restrictive temperatures show that apical fates (cotyledons plus shoot apical meristem) can be transformed to basal fates (root) as late as transition stage. As the apical pole of transition stage embryos shows both morphological and molecular characteristics of shoot development, this demonstrates that the topless 1 mutation is capable of causing structures specified as shoot to be respecified as root. Finally, our experiments fail to show a clear link between auxin signal transduction and topless-1 mutant activity: the development of the apical root in topless mutant individuals is not dependent on the activity of the predicted auxin response factor MONOPTEROS nor is the expression of DR5, a proposed 'auxin maximum reporter', expanded in the apical domain of topless embryos.
Assuntos
Proteínas de Arabidopsis , Arabidopsis/genética , Mutação , Raízes de Plantas/fisiologia , Brotos de Planta/fisiologia , Arabidopsis/fisiologia , Regulação da Expressão Gênica de Plantas , Genes Reporter , Proteínas de Homeodomínio/genética , Homozigoto , Ácidos Indolacéticos/genética , Proteínas de Plantas/genética , Sementes/genética , Temperatura , Fatores de Transcrição/genéticaRESUMO
MicroRNAs (miRNAs) are approximately 22-nucleotide noncoding RNAs that can regulate gene expression by directing mRNA degradation or inhibiting productive translation. Dominant mutations in PHABULOSA (PHB) and PHAVOLUTA (PHV) map to a miR165/166 complementary site and impair miRNA-guided cleavage of these mRNAs in vitro. Here, we confirm that disrupted miRNA pairing, not changes in PHB protein sequence, causes the developmental defects in phb-d mutants. In planta, disrupting miRNA pairing near the center of the miRNA complementary site had far milder developmental consequences than more distal mismatches. These differences correlated with differences in miRNA-directed cleavage efficiency in vitro, where mismatch scanning revealed more tolerance for mismatches at the center and 3' end of the miRNA compared to mismatches to the miRNA 5' region. In this respect, miR165/166 resembles animal miRNAs in its pairing requirements. Pairing to the 5' portion of the small silencing RNA appears crucial regardless of the mode of post-transcriptional repression or whether it occurs in plants or animals, supporting a model in which this region of the silencing RNA nucleates pairing to its target.