Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Gene expression profiling in senescence leaves from senescence-inducible LEC2 in Arabidopsis

ABSTRACT: The profiling was conducted with the 385K Microarray manufactured at NimbleGen, Inc (http://www.nimblegen.com/) designed from 30,386 genes of Arabidopsis (TAIR v.6). RNA samples of leaves of S1 (100% green leaf from 40 days plant) and S3 (more than 50% yellow leaf) senescence stages from senescence-inducible LEC2 transgenic plant, OIL21 were subjected for analysis of Arabidopsis 385K transcriptome. Expression of 3,025 genes were significantly changed with log2 ratios greater than 1.0 or less than -1.0 in S1 or S3 stage (p value <0.05), among which 1491 were up-regulated and 1534 were down regulated. Gene annotation revealed that 7.4% genes, 223 out of 3,025 changed by LEC2 were encoded transcription factors (TF). 106 TFs were up-regulated and 115 TFs were down-regulated. A total of 9 chips were used for microarray. Total RNAs were extracted from senescence leaves at S1 and S3 stages of wild-type and senescence-inducible LEC2 transgenic Arabidopsis. Experiments were triplicated.

ORGANISM(S): Arabidopsis thaliana

SUBMITTER: Yeon-Ki Kim

PROVIDER: E-GEOD-38489 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:The aim of this experiment was to decipher a transcriptional regulatory code of Arabidopsis Auxin Response Factor 2 (ARF2) and understand the molecular basis of ARF2’s role as a promoter of dark-induced senescence. A combination of computational tools and yeast one-hybrid experiments identified potential direct transcriptional regulators of ARF2 belonging to several TF families including AP2/ERFs and AREB/ABF/ABI5s. Using ARF2 promoter mutants in transcriptional activation and Y1H experiments, the binding sites of these TFs in the ARF2 promoter were identified. AP2/ERFs including ERF1, ERF15 and ORA59 transactivate the ARF2 promoter in Arabidopsis mesophyll protoplasts and this transactivation is positively regulated by ABA. Crucially, the positive effect of ABA on ERF1 transactivation of ARF2 relies on the functional ABRE motif in the ARF2 promoter and the presence of AREB/ABF/ABI5s (ABF3 and ABF4) suggesting the existence of combinatorial regulation of ARF2 by these direct regulators. ABF3 and ABF4 were unable to activate the ARF2 promoter on their own indicating the necessity of ERF1 as a coupling TF for AREB/ABF/ABI5-driven regulation of ARF2. We also show that MED25 (a subunit of the Mediator complex) is required for proper transcriptional regulation of ARF2 during senescence. med25 plants show decreased ARF2 mRNA levels during natural senescence and, similarly to arf2-6 mutants, exhibit a delayed dark-induced senescence phenotype. We show that MED25 is important for ERF1-, ERF15- and ORA59-dependent transactivation of ARF2 in Arabidopsis protoplasts and finally idenitfy genes differentially expressed in arf2-6 mutants compared to wildtype during dark-induced senescence. Hence, we elucidate a regulatory network around ARF2 identifying combinatorial transcriptional regulation of this gene during senescence as well as likely downstream targets. The combinatorial regulation, comprised of AP2/ERFs and AREB/ABF/ABI5s TFs, enables integration of at least two key hormone signaling pathways to drive expression of a key senescence regulator.

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Gene expression profiling in senescence leaves from senescence-inducible LEC2 in Arabidopsis

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