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A Local Regulatory Network Around Three NAC Transcription Factors in Stress Responses and Senescence in Arabidopsis leaves (expression between lines at timepoints).

ABSTRACT: A network model has been generated that describes the immediate gene expression cascade surrounding three similar but distinct NAC transcription factors that have roles to play in leaf senescence and in many stress responses in Arabidopsis. ANAC019, ANAC055 and ANAC072 belong to the same clade of NAC domain genes and have overlapping expression patterns. A combination of promoter DNA/protein interactions identified using yeast 1 hybrid analysis and modeling using gene expression time course data has been used to predict the regulatory network upstream of these genes. Similarities and divergence in regulation during a variety of stress responses are predicted by different combinations of upstream transcription factors binding and also by the modeling. Mutant analysis with potential upstream genes was used to test and confirm some of the predicted interactions.. Gene expression measurements in mutants of ANAC019 and ANAC055 at different times during leaf senescence have shown a distinctly different role for each of these genes.

ORGANISM(S): Arabidopsis thaliana

PROVIDER: GSE45183 | GEO | 2013/04/23

SECONDARY ACCESSION(S): PRJNA193218

REPOSITORIES: GEO

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Project description:NACs (NAM/ATAF/CUC) proteins are plant-specific transcription factors with key roles in plant development and stress responses. Here, Vitis vinifera NAC61 was selected for its functional characterization as one of the key candidates of the grape post-harvest withering process. VviNAC61 is particularly expressed at veraison and post-veraison berry stages and also shows a high expression in lignified tissues such as roots, tendrils, seeds and rachis. In leaves, VviNAC61 achieves its highest expression at senescence stage. In line with this observation, a NAC family multispecies phylogenetic tree revealed that VviNAC61 clusters with VviNAC33 and AtANAC046, both involved in regulating senescence processes. VviNAC61 was transiently overexpressed in N. benthamiana leaves, showing wilting/water-soaking symptoms correlated to low stomatal conductance, high leaf temperature, increased loss of the membrane integrity and accumulation of hydrogen peroxide and water. Transcriptomic analysis of cultivar ‘Thompson Seedless’ leaves transiently overexpressing VviNAC61 and the search of TF-bound genes by DAP-seq was allowed us to narrow down a list of VviNAC61-regulated genes. The ‘secondary metabolism’, ‘transcription factor regulation’ and ‘oxidative stress’ functional categories were found to be the most enriched among the candidate target genes. More in detail, several genes encoding for the phenylpropanoid pathway-related enzymes, several MYBs and NAC genes (including VviNAC61), were bound in their promoter region by this TF and were among the upregulated genes. Direct regulation of the stilbene synthase regulator VviMYB14, the abiotic stress-related gene DHN1b, the Botrytis cinerea susceptibility gene VviWRKY52 and the VviNAC61 self-regulation were validated through dual luciferase assays. Lastly, the upstream regulation mechanism of VviNAC61 was further investigated demonstrating that VviNAC61 is controlled by and interacts with VviNAC60, a proposed master regulator of grapevine organ maturation.

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A Local Regulatory Network Around Three NAC Transcription Factors in Stress Responses and Senescence in Arabidopsis leaves (expression between lines at timepoints).

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