Dataset Information

Two GCC boxes and AP2/ERF-domain transcription factor ORA59 in jasmonate/ethylene-mediated activation of the PDF1.2 promoter in Arabidopsis.

ABSTRACT: Plant defense against microbial pathogens depends on the action of several endogenously produced hormones, including jasmonic acid (JA) and ethylene (ET). In defense against necrotrophic pathogens, the JA and ET signaling pathways synergize to activate a specific set of defense genes including PLANT DEFENSIN1.2 (PDF1.2). The APETALA2/Ethylene Response Factor (AP2/ERF)-domain transcription factor ORA59 acts as the integrator of the JA and ET signaling pathways and is the key regulator of JA- and ET-responsive PDF1.2 expression. The present study was aimed at the identification of elements in the PDF1.2 promoter conferring the synergistic response to JA/ET and interacting with ORA59. We show that the PDF1.2 promoter was activated synergistically by JA and the ET-releasing agent ethephon due to the activity of two GCC boxes. ORA59 bound in vitro to these GCC boxes and trans-activated the PDF1.2 promoter in transient assays via these two boxes. Using the chromatin immunoprecipitation technique we were able to show that ORA59 bound the PDF1.2 promoter in vivo. Finally, we show that a tetramer of a single GCC box conferred JA/ethephon-responsive expression, demonstrating that the JA and ET signaling pathways converge to a single type of GCC box. Therefore ORA59 and two functionally equivalent GCC box binding sites form the module that enables the PDF1.2 gene to respond synergistically to simultaneous activation of the JA and ET signaling pathways.

SUBMITTER: Zarei A

PROVIDER: S-EPMC3044237 | biostudies-literature | 2011 Mar

REPOSITORIES: biostudies-literature

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Two GCC boxes and AP2/ERF-domain transcription factor ORA59 in jasmonate/ethylene-mediated activation of the PDF1.2 promoter in Arabidopsis.

Zarei Adel A Körbes Ana Paula AP Younessi Parisa P Montiel Gregory G Champion Antony A Memelink Johan J

Plant molecular biology 20110119 4-5

Plant defense against microbial pathogens depends on the action of several endogenously produced hormones, including jasmonic acid (JA) and ethylene (ET). In defense against necrotrophic pathogens, the JA and ET signaling pathways synergize to activate a specific set of defense genes including PLANT DEFENSIN1.2 (PDF1.2). The APETALA2/Ethylene Response Factor (AP2/ERF)-domain transcription factor ORA59 acts as the integrator of the JA and ET signaling pathways and is the key regulator of JA- and ...[more]

PMID: 21246258

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Project description:The experiment is aiming to analyze the effect of overexpression of the Arabidopsis At1g06160 AP2/ERF-domain transcription factor on gene expression and the involvement of the At1g06160-regulated genes in the jasmonic acid or jasmonic acid/ethylene signal transduction pathways. The experiment was performed with 12 hybridizations, using 24 samples of species [Arabidopsis thaliana], using 12 arrays of array design [Agilent Arabidopsis 3 Oligo Microarray [G4142A]], producing 12 raw data files and 0 transformed and/or normalized data files. In the first two hybridizations (#1 and #2), two independent transgenic Arabidopsis lines containing a construct carrying the Arabidopsis gene At1g06160 fused to an inducible promoter were used. This promoter is activated after the addition of the chemical estradiol to the plant medium. Addition of estradiol results in overexpression of At1g06160. For each independent line, total RNA from estradiol-treated and -untreated plants were collected and labeled with Cy3 and Cy5, respectively, and hybridization was performed for each At1g06160-inducible line. Two other hybridizations (#3 and #4) were performed using two independent transgenic Arabidopsis control lines containing a construct carrying the GUS gene fused to an inducible promoter. For each independent line, total RNA from estradiol-treated and -untreated plants were collected and labeled with Cy3 and Cy5, respectively and hybridization was performed for each GUS-inducible line. The last 8 hybridizations were performed using wild-type Arabidopsis plants. The aim was analyze the effect of the jasmonic acid hormone or the effect of a combination of the jasmonic acid and ethylene hormones on gene expression in wild-type. Hybridizations #5 and #6: wild-type Arabidopsis seedlings were treated with jasmonic acid (JA) or with the control DMSO and total RNA was collected after 8 hours. Total RNA from JA-treated and DMSO-treated plants were collected and labeled with Cy3 and Cy5, respectively, and hybridization was performed. A second hybridization was performed using independent biological samples that were treated and labeled similarly. Hybridizations #7 and #8: wild-type Arabidopsis seedlings were treated with jasmonic acid (JA) or with the control DMSO and total RNA was collected after 24 hours. Total RNA from JA-treated and DMSO-treated plants were collected and labeled with Cy3 and Cy5, respectively, and hybridization was performed. A second hybridization was performed using independent biological samples that were treated and labeled similarly. Hybridizations #9 and #10: wild-type Arabidopsis seedlings were treated with jasmonic acid (JA) and ethylene or with the control DMSO/Na-phosphate and total RNA was collected after 8 hours. Total RNA from JA/ethylene-treated and DMSO/Na-phosphate-treated plants were collected and labeled with Cy3 and Cy5, respectively, and hybridization was performed. A second hybridization was performed using independent biological samples that were treated and labeled similarly. Hybridizations #11 and #12: wild-type Arabidopsis seedlings were treated with jasmonic acid (JA) and ethylene or with the control DMSO/Na-phosphate and total RNA was collected after 24 hours. Total RNA from JA/ethylene-treated and DMSO/Na-phosphate-treated plants were collected and labeled with Cy3 and Cy5, respectively, and hybridization was performed. A second hybridization was performed using independent biological samples that were treated and labeled similarly.

Project description:BackgroundRose (Rosa chinensis) is a traditional famous flower with valuable ornamental characteristics. However, drought stress restricts its growth and development, leading to an abnormal phenotype. One of the main transcription factor (TF) protein groups in the plant kingdom are the APETALA2/ethylene-responsive factor (AP2/ERF) proteins and are potentially involved in the growth and stress responses of various plants.ResultsOur investigation mainly focused on exploring the genome of rose and thereby we discovered 135 apparent AP2/ERF TFs. Phylogenic analyses revealed that RcAP2/ERF genes are categorized into DREB, Soloist, AP2, and ERF subfamilies, and are further classified these into 17 groups, with the same as Malus domestica and Arabidopsis thaliana. The analysis of the gene structure revealed that the introns ranged from 0 to 9 in number. Pattern examination demonstrated that the RcAP2/ERF predominantly consists of typical AP2 domains, of which the 2nd motif is the most ubiquitous. Distributions of cis-acting elements indicated that members of the AP2/ERF family are frequently involved in growth and development, phytohormone and stress response in rose species. Also, the distribution mapping of the rose chromosomes indicated that AP2/ERF class genes are dispersed among all seven chromosomes. Additionally, we isolated a novel DREB A2 subgroup gene and named it RcDREB2B. Subsequently, the RcDREB2B transcript accumulation was repressed under the mild and severe drought stress in the root samples of rose. RcDREB2B was targeted to the nucleus and exhibited transactivation in yeast cells. The overexpression of RcDREB2B was found to promote sensitivity to a higher salt concentration, ABA, and PEG at the germination and post-germination stages. Twelve putative osmotic and ABA-related genes were impaired in RcDREB2B-overexpressing plants.ConclusionsThe results provide comprehensive information regarding the gene structure, phylogenic, and distribution of the rose AP2/ERF family and bring insight into the complex transcriptional gene regulation of RcAP2/ERF. Findings in this study would also contribute to further understanding of the RcDREB2B gene in rose.

Dataset Information

Two GCC boxes and AP2/ERF-domain transcription factor ORA59 in jasmonate/ethylene-mediated activation of the PDF1.2 promoter in Arabidopsis.

Publications

Two GCC boxes and AP2/ERF-domain transcription factor ORA59 in jasmonate/ethylene-mediated activation of the PDF1.2 promoter in Arabidopsis.

Similar Datasets

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