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Microarray analysis of scap1 mutant and wild type in Arabidopsis thaliana guard cell protoplast

ABSTRACT: Stomata are highly specialized organs which consist of pairs of guard cells and regulate gas and water vapor exchange in plants. While early stages of guard cell differentiation have been described and were interpreted in analogy to processes of cell type differentiation in animals, the downstream development of functional stomatal guard cells remains poorly understood. We have isolated an Arabidopsis mutant, scap1 (stomatal carpenter 1), that develops irregularly shaped guard cells and lacks the ability to control stomatal aperture, including CO2-induced stomatal closing and light-induced stomatal opening. SCAP1 was identified as a plant-specific Dof-type transcription factor expressed in maturing guard cells but not in guard mother cells. SCAP1 regulates the expression of genes encoding key elements of stomatal functioning and morphogenesis, such as a K+ channel protein, MYB60 transcription factor, and pectin methyl esterase. Consequently, ion homeostasis was disturbed in scap1 guard cells, and esterification of extracellular pectins was impaired so that the cell walls lining the pores did not mature normally. We conclude that SCAP1 regulates essential processes of stomatal guard cell maturation and functions as a key transcription factor regulating the final stages of guard cell differentiation.

ORGANISM(S): Arabidopsis thaliana

PROVIDER: GSE43964 | GEO | 2013/02/28

SECONDARY ACCESSION(S): PRJNA188235

REPOSITORIES: GEO

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Microarray analysis of scap1 mutant and wild type in Arabidopsis thaliana guard cell protoplast

Project description:Stomata are highly specialized organs which consist of pairs of guard cells and regulate gas and water vapor exchange in plants. While early stages of guard cell differentiation have been described and were interpreted in analogy to processes of cell type differentiation in animals, the downstream development of functional stomatal guard cells remains poorly understood. We have isolated an Arabidopsis mutant, scap1 (stomatal carpenter 1), that develops irregularly shaped guard cells and lacks the ability to control stomatal aperture, including CO2-induced stomatal closing and light-induced stomatal opening. SCAP1 was identified as a plant-specific Dof-type transcription factor expressed in maturing guard cells but not in guard mother cells. SCAP1 regulates the expression of genes encoding key elements of stomatal functioning and morphogenesis, such as a K+ channel protein, MYB60 transcription factor, and pectin methyl esterase. Consequently, ion homeostasis was disturbed in scap1 guard cells, and esterification of extracellular pectins was impaired so that the cell walls lining the pores did not mature normally. We conclude that SCAP1 regulates essential processes of stomatal guard cell maturation and functions as a key transcription factor regulating the final stages of guard cell differentiation. We isolated guard cell protoplasts from 4-week-old WT(Col-0) and scap1 mutant plants and extracted RNA independently. Four biological replicates were performed for each experiment.

2013-02-28 | E-GEOD-43964 | biostudies-arrayexpress

Arabidopsis Me-0 vs tetraploid Col

Project description:To investigate gene expression differences related to the regulation of stomatal aperture between two Arabidopsis accessions, Me-0 and tetraploid Col, we performed microarray analysis on guard cell protoplasts from these two accessions. We identified 1,777 probes as up-regulated and 1,706 probes as down-regulated differentially expressed genes (DEGs) in Me-0 compared with tetraploid Col (adjusted P-value < 0.001 and log 2 fold change of greater than 1 or less than -1). Using gene set analysis to functionally characterize DEGs, there were eight significant gene sets (at the level of P-value < 0.01) among the up-regulated genes and seven significant gene set among the down-regulated genes. Genes with the lowest P-value among the up-regulated genes were related to âplant-type cell wall looseningâ, an essential step in guard cell swelling. Most of the down-regulated genes were related to the defense responses for immunity, hypoxia, and ozone, which also influence stomatal responses or gas exchange. We propose that these gene expression differences may cause the difference in stomatal aperture between Me-0 and tetraploid Col. Four samples from Me-0 and three samples from tetraploid Col are used for a microarray analysis.

2016-01-14 | E-GEOD-71914 | biostudies-arrayexpress

Arabidopsis Me-0 vs tetraploid Col

Project description:To investigate gene expression differences related to the regulation of stomatal aperture between two Arabidopsis accessions, Me-0 and tetraploid Col, we performed microarray analysis on guard cell protoplasts from these two accessions. We identified 1,777 probes as up-regulated and 1,706 probes as down-regulated differentially expressed genes (DEGs) in Me-0 compared with tetraploid Col (adjusted P-value < 0.001 and log 2 fold change of greater than 1 or less than -1). Using gene set analysis to functionally characterize DEGs, there were eight significant gene sets (at the level of P-value < 0.01) among the up-regulated genes and seven significant gene set among the down-regulated genes. Genes with the lowest P-value among the up-regulated genes were related to “plant-type cell wall loosening”, an essential step in guard cell swelling. Most of the down-regulated genes were related to the defense responses for immunity, hypoxia, and ozone, which also influence stomatal responses or gas exchange. We propose that these gene expression differences may cause the difference in stomatal aperture between Me-0 and tetraploid Col.

2016-01-14 | GSE71914 | GEO

Jasmonate-mediated stomatal closure under elevated CO2 revealed by time-resolved metabolomics

Project description:Foliar stomatal movements are critical for regulating plant water loss and gas exchange. Elevated carbon dioxide (CO2) levels are known to induce stomatal closure. However, the current knowledge on CO2 signal transduction in stomatal guard cells is limited. Here we report metabolomic responses of Brassica napus guard cells to elevated CO2 using three hyphenated metabolomics platforms: gas chromatography-mass spectrometry (MS); liquid chromatography (LC)-multiple reaction monitoring-MS; and ultra-high-performance LC-quadrupole time-of-flight-MS. A total of 358 metabolites from guard cells were quantified in a time-course response to elevated CO2 level. Most metabolites increased under elevated CO2, showing the most significant differences at 10 min. In addition, reactive oxygen species production increased and stomatal aperture decreased with time. Major alterations in flavonoid, organic acid, sugar, fatty acid, phenylpropanoid and amino acid metabolic pathways indicated changes in both primary and specialized metabolic pathways in guard cells. Most interestingly, the jasmonic acid (JA) biosynthesis pathway was significantly altered in the course of elevated CO2 treatment. Together with results obtained from JA biosynthesis and signaling mutants as well as CO2 signaling mutants, we discovered that CO2-induced stomatal closure is mediated by JA signaling.

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Project description:Environmental stimuli-triggered stomatal movement is a key physiological process that regulates CO2 uptake and water loss in plants. Stomata are defined by pairs of guard cells that perceive and transduce external signals, leading to cellular volume changes and consequent stomatal aperture change. Within the visible light spectrum, red light induces stomatal opening in intact leaves. However, there has been debate regarding the extent to which red-light-induced stomatal opening arises from direct guard cell sensing of red light versus indirect responses as a result of red light influences on mesophyll photosynthesis. Here we identify conditions that result in red-light-stimulated stomatal opening in isolated epidermal peels and enlargement of protoplasts, firmly establishing a direct guard cell response to red light. We then employ metabolomics workflows utilizing gas chromatography mass spectrometry and liquid chromatography mass spectrometry for metabolite profiling and identification of Arabidopsis guard cell metabolic signatures in response to red light in the absence of the mesophyll. We quantified 223 metabolites in Arabidopsis guard cells, with 104 found to be red light responsive. These red-light-modulated metabolites participate in the tricarboxylic acid cycle, carbon balance, phytohormone biosynthesis and redox homeostasis. We next analyzed selected Arabidopsis mutants, and discovered that stomatal opening response to red light is correlated with a decrease in guard cell abscisic acid content and an increase in jasmonic acid content. The red-light-modulated guard cell metabolome reported here provides fundamental information concerning autonomous red light signaling pathways in guard cells.

2022-01-21 | MTBLS1326 | MetaboLights

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Differentiation of Arabidopsis guard cells: analysis of the networks incorporating the basic helix-loop helix transcription factor, FAMA

Project description:In plants, formation of functional stomatal guard cells is a highly regulated event so that this cell differentiation model constitutes an interesting genetic system to explore cell differentiation in response to cell cycle controllers or cell fate determinants such as transcription factors. The latest acting bHLH in the stomatal lineage circuit is FAMA that appears to be absolutely required to promote stomatal development by controlling the GMC to GC transition. In order for FAMA to trigger guard cell development, it probably initiates a gene activation cascade leading to cell differentiation and cessation of cell division. To identify and characterize FAMA inducible genes, transcriptome analysis using the Affymetrix ATH1 micro-array chip was carried out on inducible FAMA gain of function plants at 4hrs and 48 hrs post-induction.

2010-05-12 | GSE21786 | GEO

Chimeric repressors of GLKs confer tolerance to air pollutant by closing stomata in Arabidopsis

Project description:Regulation of stomatal movement is one of the effective strategies for developing resistant crops to air pollutant because stomata allows absorption of various air pollutants, such as ozone and sulfur dioxide. Transcription factor (TF) is a fascinating target of genetic manipulation for this end because TF regulates many genes simultaneously and methods of genetic manipulation are universally established. Here, we have screened transgenic Arabidopsis lines expressing chimeric repressor of TFs in high-concentration of ozone and found that the chimeric repressors of GOLDEN-LIKE1 (GLK1) and GLK2 (GLK1-SRDX and GLK2-SRDX) conferred strong tolerance to ozone. These 35S:GLK1/2-SRDX plants also showed sulfur dioxide tolerance. Their leaves showed lower rate of transpiration than wild type and a remarkable closed-stomata phenotype. The expression of the genes encoding K+ and water channels, including KAT1 and AKT1, which are involved in stomatal opening, was downregulated in 35S:GLK1-SRDX plants. Consistently, expression of GLK1-SRDX driven by the GC1 promoter, which has an activity only in guard cell, also induced closed-stomata and an ozone tolerant phenotype. On the contrary, 35S:GLK1/2 plants showed hypersensitivity to ozone and an opened-stomata phenotype. These data suggested that GLK1 and GLK2 have an ability to induce transcriptional change in guard cell and regulate stomatal movement. Our findings provide an effective tool to confer resistance to air pollutant by regulating stomatal aperture and improve crop productivity in future.

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AtMYB60

Project description:Stomatal guard-cells modulate gas exchange between the plant and the atmosphere. Regulation of transcription is emerging as an important mechanism in controlling guard cells activity. The Arabidopsis transcription factor AtMYB60, is specifically expressed in guard cells and controls stomatal movements. Opening of stomatal pores is constitutively reduced in the atmyb60-1 null allele, and water loss during drought is diminished in the mutant compared to wild type. To address the effect of the AtMYB60 disruption on global gene expression, total mRNAs, derived from atmyb60-1 and WT rosette leaves, grown in standard conditions, were hybridized to a cDNA microarray.

2005-01-10 | E-MEXP-225 | biostudies-arrayexpress

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