Transcription factor activity in cross-regulation between seedling hormone responses
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ABSTRACT: Cross-regulation between hormone signalling pathways is indispensable for plant growth and development. However, the molecular mechanisms by which multiple hormones interact and co-ordinate activity are poorly understood. Here, we generated a cross-regulation network explaining how hormone signals are integrated from multiple pathways in etiolated Arabidopsis (Arabidopsis thaliana) seedlings. To do so we comprehensively characterized transcription factor activity during plant hormone responses and reconstructed dynamic transcriptional regulatory models for six hormones; abscisic acid, brassinosteroid, ethylene, jasmonic acid, salicylic acid and strigolactone/karrikin. These models incorporated target data for hundreds of transcription factors coupled with thousands of protein-protein interactions. Each hormone recruited different combinations of transcription factors, a subset of which were shared between hormones. Hub target genes existed within hormone transcriptional networks, exhibiting transcription factor activity themselves. A group of MITOGEN-ACTIVATED PROTEIN KINASES were also identified as potential key points of cross-regulation between multiple hormones. Loss of functional MPK6 disrupted the global proteome and phosphoproteome, and transcriptome in the response to hormones. Finally, we determined that all hormones drive substantial alternative splicing that has distinct effects on the transcriptome compared with differential gene expression, acting in early hormone responses. These results provide a comprehensive understanding of the common features of plant transcriptional regulatory pathways and how cross-regulation between hormones acts upon gene expression.
Project description:Plant hormones involved in environmental stresses, namely abscisic acid (ABA), salicylic acid (SA), and jasmonic acid (JA), have been shown to interact with each other in a complex manner. To address the network of the hormone interactions, we have investigated the changes in expression under multiple hormone treatments, ABA+SA and ABA+JA. We chose cultured cells to remove the difference in the response to hormones among developmental cells or tissues. The cells were treated for 3hr and 24hr to see the rapid or transient response and steady-state response. The obtained data indicate that ABA and SA affect antagonistically, but these hormones affected many genes collaboratively. Indeed, according to the microarray data, there are many genes that responded only to ABA+SA. In addition, the ABA+SA responsive genes also responded to ABA+JA. These data suggest that hormone crosstalk is more complicated than expected and that more systematic analysis is required to untangle the hormone crosstalk network. To investigate the hormonal interactions, Arabidopsis T87 cultured cells were exposed to ABA, SA, or JA alone, or two hormones simultaneously, ABA+SA or ABA+JA, for 3hr and 24 hr. Comparing the data among those treatments, the relationships among these hormones were deduced.
Project description:Plant hormones involved in environmental stresses, namely abscisic acid (ABA), salicylic acid (SA), and jasmonic acid (JA), have been shown to interact with each other in a complex manner. To address the network of the hormone interactions, we have investigated the changes in expression under multiple hormone treatments, ABA+SA and ABA+JA. We chose cultured cells to remove the difference in the response to hormones among developmental cells or tissues. The cells were treated for 3hr and 24hr to see the rapid or transient response and steady-state response. The obtained data indicate that ABA and SA affect antagonistically, but these hormones affected many genes collaboratively. Indeed, according to the microarray data, there are many genes that responded only to ABA+SA. In addition, the ABA+SA responsive genes also responded to ABA+JA. These data suggest that hormone crosstalk is more complicated than expected and that more systematic analysis is required to untangle the hormone crosstalk network.
Project description:Plants are targets of volatile organic compounds (VOCs) released as a part of plant-plant communication, within-plant self-signaling and plant-microbe interactions. Therefore, understanding VOC perception and downstream signaling is vital for unraveling the mechanisms behind information exchange in plants, which remain largely unexplored. Using the hormone-like function of volatile terpenoids in reproductive organ development as a system with a visual marker for communication, we demonstrated that among the four petunia karrikin-insensitive receptors, PhKAI2ia stereo-specifically perceives the (-)-germacrene D signal, triggering a karrikin-like signaling cascade. This study provides new insights into plant olfaction, uncovers the role(s) of the unique intermediate clade of KAI2 receptors, illuminates the involvement of KAI2ia-dependent signaling pathway in volatile communication and gives insight into the long-standing question about the nature of potential endogenous karrikin-like ligand(s).
Project description:A time course gene expression profiling of rice treated with various plant hormones (abscisic acid, gibberelin, auxin, brassinosteroid, cytokinin and jasmonic acid) was performed to obtain an overall signature of the rice transcriptome in response to each phytohormone. The transcriptome of rice seedlings treated with various plant hormones such as abscisic acid (ABA), gibberellic acid 3 (GA3), indole-3-acetic acid (IAA), brassinolide (BL), trans-zeatin (tZ), and jasmonic acid (JA) was characterized by microarray analysis. Seven-day old seedlings were transferred in culture solutions with specific hormones and in culture solution without hormone to serve as control (mock treatment). Root samples were collected and analyzed in three replicates at pretreatment (control) and after 15 min, 30 min, 1 h, 3 h and 6 h incubation in culture solutions with each hormone. Shoot samples were collected and analyzed in two replicates at pretreatment (control) and after 1 h, 3 h, 6 h and 12 h incubation culture solutions with each hormone. A total of 93 microarray data for root and 50 microarray data for shoot were obtained.
Project description:Phytohormones are involved in diverse aspects of plant life including the regulation of plant growth, development and reproduction, as well as governing stress and defence responses. We have generated a comprehensive transcriptional reference map of the early potato responses to exogenous application of the defence hormones abscisic acid, brassinosteroid, ethylene, salicylic acid and jasmonic acid. Amongst microarray probes representing the 39000 predicted genes, a total of 3175 and 2873 probes were significantly differentially expressed at 1 h and 6 h after hormone treatment, respectively. Marker genes identified for the early hormone responses in potato include: a homeodomain 20 transcription factor (DMG400000248) for abscisic acid; a SAUR gene (DMG400016561) induced in Epibrassinolide treated plants at 6 hours; an osmotin gene (DMG400003057) specifically enhanced by the ethylene precursor aminocyclopropanecarboxylic acid; a gene weakly similar to AtWRKY40 (DMG402007388) that was induced by salicylic acid and; a jasmonate ZIM-domain protein 1 (DMG400002930) which was specifically activated by methyl jasmonate. An online database has been set up to query the expression patterns of potato genes represented on the microarray that can also incorporate future microarray or RNAseq-based expression studies.
Project description:Ethylene is a gaseous plant hormone that regulates plant growth and development. Broad reprogramming of gene expression is required for ethylene responses. The primary ethylene transcription factor (TF) ETHYLENE INSENSITIVE3 (EIN3) drives expression of secondary TFs including the ETHYLENE RESPONSE DNA-BINDING FACTORS (EDFs), but the role of the EDFs within the ethylene genome regulatory network is not understood. Here, we describe an investigation into the function of the EDFs in ethylene signalling and hormonal cross-regulation. We determined the target genes and binding dynamics of EDFs 1, 2, 3 during an ethylene response and the effects of edf1234 quadruple mutation on gene expression. The EDFs and EIN3 shared a large proportion of their target genes but had different functions. The EDFs were associated with repression of target genes, but this was superseded by activation when EIN3 bound the same genes. Genes important in other hormone signalling pathways, in particular abscisic acid (ABA), were targets of the EDFs. This demonstrates how ethylene engages hormonal cross-regulation to repress genes in competing signalling pathways and prioritize itself.
Project description:Up to now, the mechanism of the effect of topping on tobacco hormone regulation is not clear, and most studies on plant hormone signal transduction pathways rely on gene or transcriptional pathways. In this study, the regulatory mechanism of hormones in roots and leaves of topped and untopped tobacco was studied at the protein level.
Project description:The hormone jasmonic acid (JA) controls a plethora of crucially important processes in plants through a signaling pathway orchestrated by the transcription factor MYC2 and its closest relatives. Understanding the systems-level actions of transcription factors provides insight into how the genome is reprogrammed in response to environmental stimuli. However, deeper biological insight can be obtained if transcription factor activity is set in the broader regulatory context of the cell and the downstream organismal phenotypes the transcription factors control. Here, we have investigated the MYC2-governed genome regulatory network that controls JA responses in Arabidopsis thaliana etiolated seedlings. We have generated an integrated framework of the response to JA that spans from the activity of master and secondary-regulatory transcription factors, through gene expression outputs and alternative splicing to protein abundance changes, protein phosphorylation and chromatin remodeling. We have integrated time series transcriptome analysis with (phospho)proteomic data using gene regulatory network models. These enable us to predict previously unknown points of crosstalk from JA to other signaling pathways and to identify new components of the JA regulatory mechanism, which we validated through targeted mutant studies. The result is a comprehensive understanding of how a plant hormone remodels cellular function and plant behavior, the general principles of which provide a framework for analysis of cross-regulation between other hormone and stress signaling pathways.
Project description:Salicylic acid (SA) and ethylene (ET) are two important plant hormones that regulate numerous plant growth, development, and stress response processes. Previous studies have suggested functional interplay of SA and ET in defense response, but precisely how these two hormones co-regulate plant growth and development processes remains unclear. The present findings reveal an antagonism between SA and ET in apical hook formation, a process that ensures successful soil emergence of dicotyledonous etiolated seedlings. Exogenous SA inhibited the ET-induced expression of HOOKLESS1 (HLS1) in a manner dependent on ETHYLENE INSENSITIVE3 (EIN3)/EIN3-LIKE1 (EIL1), the core transcription factors in the ET signaling pathway. We found that SA-activated NONEXPRESSER OF PR GENES1 (NPR1) physically interacted with EIN3 and interfered with the direct binding of EIN3 to target gene promoters, including the HLS1 promoter. Transcriptomic analysis further revealed that NPR1 and EIN3/EIL1 coordinately regulated subsets of genes that mediate plant growth and stress responses, suggesting that the interaction between NPR1 and EIN3/EIL1 might be an important mechanism for integrating the SA and ET signaling pathways in multiple physiological processes. Taken together, our findings shed light on the molecular mechanism underlying SA regulation of apical hook formation as well as the antagonism between SA and ET in early seedling establishment and possibly other physiological processes.