RESUMEN
SHORT-INTERNODES/STYLISH (SHI/STY)-family proteins redundantly regulate development of lateral organs in Arabidopsis thaliana. We have previously shown that STY1 interacts with the promoter of the auxin biosynthesis gene YUCCA (YUC)4 and activates transcription of the genes YUC4, YUC8 and OCTADECANOID-RESPONSIVE ARABIDOPSIS AP2/ERF (ORA)59 independently of protein translation. STY1 also affects auxin levels and auxin biosynthesis rates. Here we show that STY1 induces the transcription of 16 additional genes independently of protein translation. Several of these genes are tightly co-expressed with SHI/STY-family genes and/or down-regulated in SHI/STY-family multiple mutant lines, suggesting them to be regulated by SHI/STY proteins during plant development. The majority of the identified genes encode transcription factors or cell expansion-related enzymes and functional studies suggest their involvement in stamen and leaf development or flowering time regulation.
Asunto(s)
Proteínas de Arabidopsis/fisiología , Arabidopsis/genética , Arabidopsis/fisiología , Proteínas Portadoras/fisiología , Genes de Plantas , Arabidopsis/crecimiento & desarrollo , Proteínas de Arabidopsis/genética , Secuencia de Bases , Proteínas Portadoras/genética , Proliferación Celular , ADN de Plantas/genética , Flores/crecimiento & desarrollo , Regulación del Desarrollo de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Técnicas de Inactivación de Genes , Ácidos Indolacéticos/metabolismo , Mutación , Oxigenasas/genética , Oxigenasas/fisiología , Plantas Modificadas Genéticamente , Transactivadores/genética , Transactivadores/fisiología , Factores de Transcripción/genética , Factores de Transcripción/fisiologíaRESUMEN
Auxin/indole-3-acetic acid (IAA) biosynthesis in Arabidopsis (Arabidopsis thaliana) plays a major role in growth responses to developmental and genetic signals as well as to environmental stimuli. Knowledge of its regulation, however, remains rudimentary, and few proteins acting as transcriptional modulators of auxin biosynthesis have been identified. We have previously shown that alteration in the expression level of the SHORT INTERNODES/STYLISH (SHI/STY) family member STY1 affects IAA biosynthesis rates and IAA levels and that STY1 acts as a transcriptional activator of genes encoding auxin biosynthesis enzymes. Here, we have analyzed the upstream regulation of SHI/STY family members to gain further insight into transcriptional regulation of auxin biosynthesis. We attempted to modulate the normal expression pattern of STY1 by mutating a putative regulatory element, a GCC box, located in the proximal promoter region and conserved in most SHI/STY genes in Arabidopsis. Mutations in the GCC box abolish expression in aerial organs of the adult plant. We also show that induction of the transcriptional activator DORNRÖSCHEN-LIKE (DRNL) activates the transcription of STY1 and other SHI/STY family members and that this activation is dependent on a functional GCC box. Additionally, STY1 expression in the strong drnl-2 mutant or the drn drnl-1 puchi-1 triple mutant, carrying knockdown mutations in both DRNL and its close paralogue DRN as well as one of their closest homologs, PUCHI, was significantly reduced, suggesting that DRNL regulates STY1 during normal plant development and that several other genes might have redundant functions.
Asunto(s)
Proteínas de Arabidopsis/metabolismo , Arabidopsis/genética , Regulación de la Expresión Génica de las Plantas/genética , Ácidos Indolacéticos/metabolismo , Motivos de Nucleótidos/genética , Reguladores del Crecimiento de las Plantas/metabolismo , Aminoácidos Cíclicos/farmacología , Arabidopsis/crecimiento & desarrollo , Arabidopsis/fisiología , Proteínas de Arabidopsis/genética , Secuencia de Bases , Proteínas Portadoras/genética , Proteínas Portadoras/metabolismo , Cotiledón/genética , Cotiledón/crecimiento & desarrollo , Cotiledón/fisiología , Mutación , Componentes Aéreos de las Plantas/genética , Componentes Aéreos de las Plantas/crecimiento & desarrollo , Componentes Aéreos de las Plantas/fisiología , Raíces de Plantas/genética , Raíces de Plantas/crecimiento & desarrollo , Raíces de Plantas/fisiología , Plantas Modificadas Genéticamente , Regiones Promotoras Genéticas/genética , Plantones/genética , Plantones/crecimiento & desarrollo , Plantones/fisiología , Transducción de Señal , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Transcripción GenéticaRESUMEN
The plant hormone auxin plays fundamental roles in vascular plants. Although exogenous auxin also stimulates developmental transitions and growth in non-vascular plants, the effects of manipulating endogenous auxin levels have thus far not been reported. Here, we have altered the levels and sites of auxin production and accumulation in the moss Physcomitrella patens by changing the expression level of homologues of the Arabidopsis SHI/STY family proteins, which are positive regulators of auxin biosynthesis genes. Constitutive expression of PpSHI1 resulted in elevated auxin levels, increased and ectopic expression of the auxin response reporter GmGH3pro:GUS, and in an increased caulonema/chloronema ratio, an effect also induced by exogenous auxin application. In addition, we observed premature ageing and necrosis in cells ectopically expressing PpSHI1. Knockout of either of the two PpSHI genes resulted in reduced auxin levels and auxin biosynthesis rates in leafy shoots, reduced internode elongation, delayed ageing, a decreased caulonema/chloronema ratio and an increased number of axillary hairs, which constitute potential auxin biosynthesis sites. Some of the identified auxin functions appear to be analogous in vascular and non-vascular plants. Furthermore, the spatiotemporal expression of the PpSHI genes and GmGH3pro:GUS strongly overlap, suggesting that local auxin biosynthesis is important for the regulation of auxin peak formation in non-vascular plants.
Asunto(s)
Arabidopsis/genética , Briófitas/genética , Arabidopsis/anatomía & histología , Arabidopsis/crecimiento & desarrollo , Southern Blotting , Briófitas/enzimología , Briófitas/crecimiento & desarrollo , Sistema Enzimático del Citocromo P-450/genética , ADN de Plantas/genética , ADN de Plantas/aislamiento & purificación , Amplificación de Genes , Técnicas de Inactivación de Genes , Genoma de Planta , Ácidos Indolacéticos/metabolismo , Fenotipo , Reguladores del Crecimiento de las Plantas/genética , Reguladores del Crecimiento de las Plantas/metabolismo , Plantas Modificadas Genéticamente/genética , Reacción en Cadena de la Polimerasa/métodosRESUMEN
The establishment and maintenance of auxin maxima in vascular plants is regulated by auxin biosynthesis and polar intercellular auxin flow. The disruption of normal auxin biosynthesis in mouse-ear cress (Arabidopsis thaliana) leads to severe abnormalities, suggesting that spatiotemporal regulation of auxin biosynthesis is fundamental for normal growth and development. We have shown previously that the induction of the SHORT-INTERNODES/STYLISH (SHI/STY) family member STY1 results in increased transcript levels of the YUCCA (YUC) family member YUC4 and also higher auxin levels and auxin biosynthesis rates in Arabidopsis seedlings. We have also shown previously that SHI/STY family members redundantly affect development of flowers and leaves. Here, we further examine the function of STY1 by analyzing its DNA and protein binding properties. Our results suggest that STY1, and most likely other SHI/STY members, are DNA binding transcriptional activators that target genes encoding proteins mediating auxin biosynthesis. This suggests that the SHI/STY family members are essential regulators of auxin-mediated leaf and flower development. Furthermore, the lack of a shoot apical meristem in seedlings carrying a fusion construct between STY1 and a repressor domain, SRDX, suggests that STY1, and other SHI/STY members, has a role in the formation and/or maintenance of the shoot apical meristem, possibly by regulating auxin levels in the embryo.
Asunto(s)
Proteínas de Arabidopsis/fisiología , Proteínas Portadoras/fisiología , Ácidos Indolacéticos/metabolismo , Transactivadores/metabolismo , Secuencia de Aminoácidos , Arabidopsis/genética , Arabidopsis/fisiología , Proteínas de Arabidopsis/química , Proteínas Portadoras/química , Datos de Secuencia Molecular , Regiones Promotoras Genéticas , Homología de Secuencia de AminoácidoRESUMEN
In angiosperms, the gynoecium constitutes the female reproductive organ that after fertilization develops into a fruit and in Arabidopsis thaliana the gynoecium is formed by the congenital fusion of two carpels. In the last few years many genes involved in female organ development have been identified and there have been several reports on the involvement of the plant hormone auxin in gynoecium patterning. An auxin gradient has been suggested to establish the apical-basal patterning of the gynoecium and recently it has been shown that elevated apical auxin levels can compensate for the loss of several style-promoting factors but that auxin is dependent on their action in apical-basal patterning. Here we discuss the role of auxin and different upstream, downstream or parallel factors in the apical-basal patterning of the gynoecium. We focus specifically on the development of style and stigma and discuss the most recent findings.
Asunto(s)
Arabidopsis/crecimiento & desarrollo , Flores/crecimiento & desarrollo , Ácidos Indolacéticos/metabolismo , Arabidopsis/genética , Regulación de la Expresión Génica de las Plantas , Genes de Plantas , Factores de Transcripción/genéticaRESUMEN
Patterning of the Arabidopsis thaliana gynoecium is dependent on the localization and concentration of the plant hormone auxin and it has been previously reported that STYLISH1 (STY1) activates transcription of the auxin biosynthesis gene YUCCA4 (YUC4) and affects gynoecium development. Here, the relationship between auxin, STY1 and other regulators of gynoecium development was examined. Exogenous auxin in droplets of lanolin paste were applied to young gynoecia; auxin biosynthesis rate was measured and STY1 overexpression or chemically mediated polar auxin transport (PAT) inhibition were induced in various mutants. The style phenotype of sty1-1sty2-1 mutants was restored by exogenous application of auxin, and STY1 over-activation resulted in an elevated auxin biosynthesis rate. Both over-activation of STY1 and inhibition of PAT restored the stylar defects of several unrelated mutants, but with regard to gynoecium apical-basal patterning the mutants responded differently to inhibition of PAT. These results suggest that reduced auxin concentrations cause the sty1-1 sty2-1 phenotype, that STY1 induces auxin biosynthesis, that elevated apical auxin concentrations can compensate for the loss of several style-promoting factors, and that auxin may act downstream of, or in parallel with these during style development but is dependent on their action in apical-basal patterning.
Asunto(s)
Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/fisiología , Arabidopsis/crecimiento & desarrollo , Proteínas Portadoras/genética , Proteínas Portadoras/fisiología , Flores/crecimiento & desarrollo , Ácidos Indolacéticos/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Flores/genética , Regulación de la Expresión Génica de las Plantas , Morfogénesis/genética , Morfogénesis/fisiología , Mutación , Fenotipo , Factores de Transcripción , Transcripción GenéticaRESUMEN
Compared to wood, cell suspension cultures provide convenient model systems to study many different cellular processes in plants. Here we have established cell suspension cultures of Populus tremula L. x P. tremuloides Michx. and characterized them by determining the enzymatic activities and/or mRNA expression levels of selected cell wall-specific proteins at the different stages of growth. While enzymes and proteins typically associated with primary cell wall synthesis and expansion were detected in the exponential growth phase of the cultures, the late stationary phase showed high expression of the secondary-cell-wall-associated cellulose synthase genes. Interestingly, detergent extracts of membranes from aging cell suspension cultures exhibited high levels of in vitro cellulose synthesis. The estimated ratio of cellulose to callose was as high as 50 : 50, as opposed to the ratio of 30 : 70 so far achieved with membrane preparations extracted from other systems. The increased cellulose synthase activity was also evidenced by higher levels of Calcofluor white binding in the cell material from the stationary-phase cultures. The ease of handling cell suspension cultures and the improved capacity for in vitro cellulose synthesis suggest that these cultures offer a new basis for studying the mechanism of cellulose biosynthesis.