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Direct target gene analyses for TCP20 using a glucocorticoid inducible TCP20 (TCP20-GR)

ABSTRACT: TCP (TEOSINTE BRANCHED1/CYCLOIDEA/PCF1) transcription factors control developmental processes in plants. We identified direct target genes of the Arabidopsis class I TCP20 protein in leaf development based on a glucocorticoid receptor induction assay and genome-wide expression studies. For this, we tagged TCP20 with a glucucorticoid receptor (GR) domain and transformed the resulting TCP20-GR construct into tcp20 knockout plants. Induction of these and wild type controls was done with Dexamethasone and Cycloheximide. Plants were harvested 8 h after induction, uninduced plants were taken as control.

ORGANISM(S): Arabidopsis thaliana

PROVIDER: GSE29012 | GEO | 2011/08/09

SECONDARY ACCESSION(S): PRJNA140465

REPOSITORIES: GEO

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Direct target gene analyses for TCP20 using a glucocorticoid inducible TCP20 (TCP20-GR)

Project description:TCP (TEOSINTE BRANCHED1/CYCLOIDEA/PCF1) transcription factors control developmental processes in plants. We identified direct target genes of the Arabidopsis class I TCP20 protein in leaf development based on a glucocorticoid receptor induction assay and genome-wide expression studies. For this, we tagged TCP20 with a glucucorticoid receptor (GR) domain and transformed the resulting TCP20-GR construct into tcp20 knockout plants. Induction of these and wild type controls was done with Dexamethasone and Cycloheximide. Plants were harvested 8 h after induction, uninduced plants were taken as control. In sum, 8 samples, each pools of 30 seedlings. Wild type and TCP20-GR plants at induction times t=0 and t=8, in two biological replicates.

2011-08-08 | E-GEOD-29012 | biostudies-arrayexpress

The MPK8-TCP14 pathway promotes seed germination in Arabidopsis

Project description:Accurate control of dormancy release and germination is critical for successful plantlet establishment. In this study, we characterized mutants of Arabidopsis MAP kinase 8 (MPK8) presenting altered dormancy. We identified Teosinte Branched1/Cycloidea/Proliferating cell factor 14 (TCP14), a key transcription factor for germination, as a partner of MPK8 and investigated the processes controlled by MPK8-TCP14 pathway. Kinase assay was performed using immunoprecipitated MPK8-HA protein and recombinant GST-TCP14 protein as target to find TCP14 MPK8-specific phosphorylation sites.

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BRANCHED1-like gene CsTCP18a contributes to shoot branching and leaf development in cucumber

Project description:Previous study we have reported the cucumber TEOSINTE BRANCHED1/CYCLOIDEA/PCF (TCP) family gene BRANCHED1 (CsBRC1) as a main transcription factor functions to regulate shoot branching. Here, we found CsBRC1 (CsTCP18b in this study) had a paralogous gene CsTCP18a. To investigate whether the function of CsTCP18a was same as CsTCP18b, we carried out biochemical experiments and genetic transformation. The Real-Time PCR and in situ hybridization showed that CsTCP18a displayed different expression patterns in cucumber compared with CsTCP18b. Ectopic expression of CsTCP18a in Arabidopsis tcp18 (brc1) mutants resulted in a decreased number of rosette branches and rosette leaves, whereas silencing CsTCP18a in cucumber only led to a deformed true leaf of seedling without influencing the shoot branching. RNA-seq analysis of wild-type plants and CsTCP18a-RNAi lines implicated that CsTCP18a regulated early leaf development of cucumber through affecting the transcripts of auxin and cytokinin related genes. Further studies indicated that CsTCP18a could directly interact with CsTCP10 and CsTCP18b in vitro and in vivo. Therefore, our data suggested that CsTCP18a had functional redundancy with CsTCP18b in inhibiting axillary buds outgrowth, and it could also regulate leaf development during cucumber seedling.

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Rice transcriptional repressor OsTIE1 controls anther dehiscence and male sterility by regulating JA biosynthesis

Project description:The coordination of pollen viability, stamen filament elongation and anther dehiscence is essential for the successful pollination and reproduction in angiosperms. The hormone jasmonic acid (JA) are crucial for these processes. However, the tight regulation of JA biosynthesis during stamen development is still largely unknown. Here, we demonstrate that the rice (Oryza sativa) ERF-associated amphiphilic repression (EAR) motif-containing protein TCP INTERACTOR CONTAINING EAR MOTIF PROTEIN1 (OsTIE1) tightly regulates JA biosynthesis by repressing TEOSINTE BRANCHED1/CYCLOIDEA/PROLIFERATINGCELLFACTORS (TCP) transcription factor OsTCP1/PCF5 during stamen development. The loss of OsTIE1 function in ostie1 mutants causes male sterility. The ostie1 mutants display inviable pollen, the early stamen filament elongation and the precocious anther dehiscence. JA biosynthesis is activated earlier and JA level is precociously increased in anthers of ostie1. The expression of OsTIE1 is developmentally regulated during stamen development. OsTIE1 is localized in nuclei and has transcriptional repression activity. We further show that OsTIE1 directly interacts with OsTCP1, and overexpression of OsTCP1 caused early anther dehiscence resembling that of ostie1. The expression of genes encoding key enzymes of JA biosynthesis including Oryza sativa lipoxygenase (OsLOX) genes is regulated by the OsTIE1-OsTCP1 complex. Our findings discover that the OsTIE1-OsTCP1 module plays critical roles in stamen development by finely tuning JA biosynthesis, and provides a new foundation to create male sterile plants for hybrid seed production.

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tt1-gr-TT1 gene network

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2019-12-18 | GSE134014 | GEO

Transcription profiling of Arabidopsis plants inducibly expressing LEC2:GR or ttg:GR to identify primary and secondary targets of the LEC2 transcription factor

Project description:Comparison of various inductibles constructions of arabidopsis treated by differents solutions. The aim of this project is to determine the primary and secondary targets of the LEC2 transcrition factor. The induction by the dexamethasone allows the expression of constructions LEC2:GR or ttg:GR introduced into transforming plants.The dexamethasone induces the expression of the contruct which carries the receptor to the glucocorticoïde (GR.). The cycloheximide is a inhibitor of the translation and allows, coupled with the dexamethasome, the identification of the primary targets only. The LEC2_GR25 and LEC2_GR31 constructs are two independents transforming plants of the cDNA-LEC2_GR construct. Both were used in order to have an idea of the variability of the data transcriptome obtained. The ttg_GR construct is used here as control ; ttg is another transcription factor, thus if induction with the dexamethasone works well, the targets induced in the two mutants will be different.

2007-01-01 | E-MEXP-807 | biostudies-arrayexpress

CsBRC1 inhibits axillary bud outgrowth by repressing auxin efflux carriers CsPIN1b and CsPIN3 and in response to shade in cucumber

Project description:Shoot branching is an important agronomic trait that directly determines plant architecture and affects crop productivity. To promote crop yield and quality, axillary branches need to be manually removed during cucumber production for fresh market and thus are undesirable. Auxin is well known as the primary signal imposing for apical dominance and acts as a repressor for branching outside of the lateral buds. The TEOSINTE BRANCHED1/CYCLOIDEA /PCF (TCP) family gene BRANCHED1 (BRC1) has been shown to be the central integrator for multiple environmental and developmental factors that functions locally to inhibit shoot branching. However, no direct link has been reported between auxin and BRC1. Here, we find that cucumber BRANCHED1 (CsBRC1) is expressed in the axillary buds and displays higher expression level in cultivated cucumber than its wild ancestor. Knockdown of CsBRC1 by RNAi leads to increased bud outgrowth and reduced auxin accumulation in buds. We further show that CsBRC1 directly binds to two auxin efflux carriers PIN-FORMED (CsPIN1b and CsPIN3) and negatively regulates their expression. Ectopic expression of CsPIN1b or CsPIN3 driven by CsBRC1 promoter results in increased shoot branching. Moreover, shade induces the expression of CsBRC1 in cultivated cucumber, but not in wild ancestor, which may be partly due to the addition of two light response elements in the CsBRC1 promoter of cultivated cucumber. Therefore, our data suggest the formation of a regulatory pathway of shade-CsBRC1-auxin transport in suppressing lateral bud outgrowth during cucumber domestication.

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Transcription profiling of TCP10m-GR plants

Project description:Transgenic plants expressing a microRNA insensitive version of TCP10 (TCP10m) fused to the Glucocortocoid receptor (GR) in the jaw-D background and jaw-D mutants were grown for 21 days on MS agar. Nuclear import of TCP10m-GR was induced by transfer of plantlets to MS medium containing 10M-5M Dexamethasone and 10 M-5M Cycloheximide. The above ground parts of whole plantlets were harvested after 0, 2 and 4 hours.

2013-10-25 | E-TABM-1191 | biostudies-arrayexpress

LsAP2 regulates leaf morphology by inhibiting CIN-like TCP transcription factors and repressing LsKAN2 in lettuce

Project description:Leaf size and flatness directly affect photosynthesis and are closely related to agricultural yield. The final leaf size and shape are coordinately determined by cell proliferation, differentiation, and expansion during leaf development. Lettuce (Lactuca sativa L.) is one of the most important leafy vegetables worldwide, and lettuce leaves vary in shape and size. However, the molecular mechanisms of leaf development in lettuce are largely unknown. In this study, we showed that the lettuce APETALA2 (LsAP2) gene regulates leaf morphology. LsAP2 encodes a transcriptional repressor that contains the conserved EAR motif, which mediates interactions with the TOPLESS/TOPLESS-RELATED (TPL/TPR) corepressors. Overexpression of LsAP2 led to small and crinkly leaves, and many bulges were seen on the surface of the leaf blade. LsAP2 physically interacted with the CINCINNATA (CIN)-like TEOSINTE BRANCHED1/CYCLOIDEA/PROLIFERATING CELL FACTOR (TCP) transcription factors and inhibited their transcriptional activation activity. RNA sequencing analysis showed that LsAP2 affected the expression of auxin- and polarity-related genes. In addition, LsAP2 directly repressed the abaxial identity gene KANADI2 (LsKAN2). Together, these results indicate that LsAP2 regulates leaf morphology by inhibiting CIN-like TCP transcription factors and repressing LsKAN2, and our work provides insights into the regulatory mechanisms of leaf development in lettuce.

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Ath_Agilent_v2_Riechmann array expression profile AP1-GR ap1cal inflorescences - chx experiment

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