Project description:Arabidopsis overexpressing ERF96 are more resistant against necrotrophic pathogen such as Botrytis cinerea. ERF96 is a member of the AP2/ERF superfamily of transcription factor. We used Microarray to determine which gene expressions are up-regulated due to an accumulation of ERF96 and to identify the putative direct target of this transcription factor.
Project description:Arabidopsis overexpressing ERF96 are more resistant against necrotrophic pathogen such as Botrytis cinerea. ERF96 is a member of the AP2/ERF superfamily of transcription factor. We used Microarray to determine which gene expressions are up-regulated due to an accumulation of ERF96 and to identify the putative direct target of this transcription factor. Microarray were performed from the RNA of a four-week old Arabidopsis overexpressing ERF96 and WT(col-0). RNA was extracted during three independant experiments
Project description:Primary cell wall is an essential cell structure for plant playing major roles in plant growth, differentiation, and stress responses. Here we demonstrate that a group of AP2-ERF transcription factor regulates primary cell wall formation and can induce massive accumulation of it in empty fiber cell of the nst1-1 nst3-1 mutant lacking secondary cell wall in Arabidopsis. The transgenic plants expressing one of the AP2-ERF transcription factors fused with VP16 transcriptional activation domain under the control of NST3 promoter in the nst1-1 nst3-1 mutant showed similar level of cell wall contents with wild type by the massive accumulation of cell wall which lacks lignin and xylan. The transgenic plants showed 70% higher saccharification efficiency than wild type. Gene expression analysis using microarray revealed that genes related to primary cell wall were highly upregulated in the transgenic plant. Moreover, chimeric-activator of the AP2-ERF transcription factor accelerated cell wall regeneration of mesophyll protoplast of Arabidopsis while the chimeric-repressor retarded it. These data suggest that the group of AP2-ERF transcription factor is key regulator of the primary cell wall formation in plant and could be employed to produce massive cell wall with readily extractable feature.
Project description:Comparison of rosette leaves of two different RAP2.2 overexpressing lines with wild type leaves. The AP2/EREBP transcription factor RAP2.2 was shown to bind to a cis-acting motif within the phytoene synthase promoter from Arabidopsis. To investigate effects of increased RAP2.2 levels, two RAP2.2 overexpressing Arabidopsis thaliana (ecotype Wassilewskija) lines were generated: one line, nosr2, carried the nos promoter and showed a two-fold increase in RAP2.2 transcript level, the second line, cmr-5, carried four copies of the CaMV-35S enhancer and showed a 12-fold increase. However, neither weak nor strong increase in RAP2.2 transcript amounts had any effect on RAP2.2 protein levels as shown by Western blot analysis. The strong robustness of RAP2.2 protein levels towards transcriptional changes can be explained by specific protein degradation which includes SINAT2, an E3 ubiquitin ligase which was isolated using a two-hybrid approach. Accordingly, global gene expression analysis using both RAP2.2 overexpressing lines showed only minor transcriptional changes which are probably due to minor growth variation than to mechanisms involved in the down-regulation of RAP2.2 protein amounts.
Project description:The AP2/ERF domain transcription factor ABSCISIC ACID INSENSITIVE4 (ABI4) modulates diverse developmental and physiological processes by integrating multiple environmental factors and phytohormone signals in plants. To further investigate the molecular mechanism of ABI4 in plant development, we examined the global gene expression in the WT and abi4-1 plants using RNA sequencing.
Project description:The genome of Arabidopsis contains 122 ERF transcription factor genes, some of which play diverse roles in the development and stress responses. A previous transcriptome analysis of Arabidopsis revealed that gene expression under combinatory stress conditions is not predictable from the combination of individual stresses (GSE46760). Searching for commonly regulated genes verified that only 11 genes were regulated in all stress combinations . Among these 11 genes, Rap2.9 (AT4G06746) was down-regulated under all stress conditions. Transgenic Arabidopsis thaliana lines overexpressing Rap2.9 under CaMV 35S promotor were generated, and the microarray analysis revealed that overexpression of Rap2.9 in A. thaliana plants causes massive reprogramming of transcription resulting in the alteration of the hormonal homeostasis and imbalances in signalling cascades related to stress response. Further analysis also revealed that Rap2.9 played an crucial role in plant development and stress tolerance in Arabidopsis plants.