Project description:Environmental stimuli, including elevated CO2, regulate stomatal development1-3 but the key mechanisms mediating the perception and relay of the CO2 signal to the stomatal development machinery remain elusive. To adapt CO2 intake to water loss, plants regulate the development of stomatal gas exchange pores in the aerial epidermis. Diverse plant species show a decrease in stomatal density in response to the continuing rise of atmospheric CO2 4. To date, one mutant, hic5, defective in cell wall wax biosynthesis, has been identified that exhibits a de-regulation of this CO2-controlled stomatal development response. Here we show that recently isolated Arabidopsis thaliana carbonic anhydrase double mutant plants6 exhibit an inversion in their response to elevated CO2, showing increased stomatal development at elevated CO2 levels. We have characterized the mechanisms mediating this response and demonstrate extracellular signaling in the regulation of CO2-controlled stomatal development by carbonic anhydrases. Transcriptomic RNA-Seq analyses show that the extracellular pro-peptide gene EPF2 7,8, but not EPF1 9, is induced at elevated CO2 in wild type, but not ca1ca4 mutant leaves. Moreover, EPF2 is essential for CO2 control of stomatal development. Using cell wall proteomic and CO2-dependent transcriptome analyses, we have identified a novel, CO2-induced extracellular protease, CRSP (CO2 Response Secreted Protease), as a mediator of CO2 controlled stomatal development. Our results identify mechanisms and genes that function in the repression of stomatal development in leaves during atmospheric CO2 elevation, including the CA1/CA4 carbonic anhydrases and the secreted protease CRSP that cleaves the pro-peptide EPF2, which in turn represses stomatal development.
Project description:Growth daylength, ambient CO2 level, and intracellular hydrogen peroxide (H2O2) availability all impact plant function by modulating signalling pathways, but interactions between them remain unclear. Using a whole-genome transcriptomics approach, we exploited the conditional photorespiratory nature of the catalase-deficient cat2 mutant to identify gene expression patterns responding to these three factors. Arabidopsis Col-0 and cat2 grown for 5 weeks in high CO2 in short days (SD) were transferred to air in SD or long days (LD), and microarray analysis was performed. Of more than 500 genes differentially expressed in Col-0 between high CO2 and transfer to air in SD, the response of about one-third was attenuated by transfer to air in LD. H2O2-responsive genes in cat2 were highly dependent on daylength. The majority of H2O2-induced genes were more strongly up-regulated after transfer to air in SD than to LD, while a smaller number showed an opposing pattern. Responses of other H2O2-dependent genes indicate redox-modulation of the daylength control of fundamental cell processes. The overall analysis provides evidence that (1) CO2 level modulates stress-associated gene expression; (2) both CO2 and H2O2 interact with daylength and photoreceptor signalling pathways; and (3) cellular signalling pathways may be primed to respond to increased H2O2 in a daylength-determined manner. Two genotypes x five conditions experiment, including Arabidopsis Col0WT and cat2-1 plants grown in soil for 5 weeks in a 8h light/16h dark (short day) regime at high CO2 concentration (3000 ppm CO2) and subsequently transferred to air (400 ppm CO2) in a short day or long day (16h light/8h dark) regime for 2 and 4 days. Three biological replicates were used that consist each of a pool of two leaves from different plants. Each sample was hybridized to one Genechip® Arabidopsis ATH1 Genome Array (Affymetrix).
Project description:Transcript profiling analysis of AtFBP7 mutant seedlings compared to wild type using Arabidopsis ATH1 GeneChip array. Keywords: 5 day old light grown seedlings, wild type and mutant
Project description:geLC-MS/MS analysis was performed to identify proteins that are present in apoplastic fluid isolated from rosette leaves of 8-week-old wild type and carbonic anhydrase (ca1ca4) mutant Arabidopsis. Plants were grown in either low (150 ppm) or high (500 ppm) CO2.
Project description:Transcript profiling analysis of vfb (Vier F-Box) mutant seedlings compared to wild type using Arabidopsis ATH1 GeneChip array. Keywords: 10 day old light grown seedlings, wild type and mutant
Project description:Transcript profiling analysis of csn4-1 light grown mutant seedlings compared to wild type using Arabidopsis ATH1 GeneChip array Keywords: 7 day old light grown seedlings, wild type and mutant
Project description:Growth daylength, ambient CO2 level, and intracellular hydrogen peroxide (H2O2) availability all impact plant function by modulating signalling pathways, but interactions between them remain unclear. Using a whole-genome transcriptomics approach, we exploited the conditional photorespiratory nature of the catalase-deficient cat2 mutant to identify gene expression patterns responding to these three factors. Arabidopsis Col-0 and cat2 grown for 5 weeks in high CO2 in short days (SD) were transferred to air in SD or long days (LD), and microarray analysis was performed. Of more than 500 genes differentially expressed in Col-0 between high CO2 and transfer to air in SD, the response of about one-third was attenuated by transfer to air in LD. H2O2-responsive genes in cat2 were highly dependent on daylength. The majority of H2O2-induced genes were more strongly up-regulated after transfer to air in SD than to LD, while a smaller number showed an opposing pattern. Responses of other H2O2-dependent genes indicate redox-modulation of the daylength control of fundamental cell processes. The overall analysis provides evidence that (1) CO2 level modulates stress-associated gene expression; (2) both CO2 and H2O2 interact with daylength and photoreceptor signalling pathways; and (3) cellular signalling pathways may be primed to respond to increased H2O2 in a daylength-determined manner.
Project description:Transcript profiling analysis of Hydraulic conductivity of Root 1 (HCR1) mutant compared to wild type (Col-0) using ARABIDOPSIS GENE1.1ST ARRAY STRIP (901793, Affymetrix, Santa Clara, USA).