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Global gene expression analysis of Arabidopsis Col-0 under normal and low oxygen conditions

ABSTRACT: Global gene expression was compared between root RNA samples from three-week-old Arabidopsis Col-0 plants subjected to 0.1% oxygen (balance nitrogen) or ambient atmospheric conditions.

ORGANISM(S): Arabidopsis thaliana

PROVIDER: GSE21504 | GEO | 2010/04/27

SECONDARY ACCESSION(S): PRJNA126005

REPOSITORIES: GEO

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Project description:Atmospheric CO2 concentrations can determine the number of stomata that form on plant leaves (Woodward & Kelly 1995 New Phyt 131: 311-327). The majority of species exhibit reduced stomatal densities at elevated CO2. However, not all plant species react in the same way to elevated CO2 levels and there is a spectrum of effects: Some species increase stomatal densities, some decrease stomatal densities, and some are unaffected. In addition to which, other environmental factors influence the number of stomata that a plant form. Light intensity has also been shown to affect stomatal numbers in various Arabidopsis ecotypes (Schluter et al. 2003 J Exp Bot 54 (383): 867-874; Lake et al. 2002 J Exp Bot 53 (367): 183-193), by increasing stomatal numbers with increasing light levels. There are many changes in gene expression under elevated CO2 conditions, so pinpointing specific genes involved in the stomatal response to CO2 is difficult. In addition, if there is crosstalk between the various signalling pathways affecting ultimate stomatal numbers this complicates further the task of finding genes specifically involved the stomatal response to CO2. Therefore we propose to look at the interaction of two known influences on stomatal numbers, light and CO2, on one specific ecotype, Col-0. We aim to test the hypothesis that light signals interact the CO2 signals that affect stomatal development. Arabidopsis thaliana Columbia-0 ecotype has previously been shown to decrease stomatal numbers in response to a doubling of ambient CO2 concentrations. Col-0 has also been shown to increase stomatal numbers in response to high light intensities. Therefore we propose to grow A. thaliana Col-0 at three light intensities (50 mmol m-2 s-1, 150 mmol m-2 s-1 and 250 mmol m-2 s-1), in both ambient and elevated (double ambient) atmospheric CO2 concentrations. By looking in more detail at how gene expression differs between plants grown at ambient and elevated CO2 at the same light intensities, and also how gene expression differs between plants grown at the same CO2 concentration but different light intensities, we aim to identify those genes involved in the stomatal developmental response to CO2 and whether genes involved in the light response can also be isolated.

Project description:Global gene expression was compared between Arabidopsis lines with altered expression of ANAC102 (over-expressed and knocked-out) and wild-type. ANAC102 is a putative NAC domain transcription factor. Gene expression was compared between an ANAC102 over-expressing line and parental ecotype C24 under ambient atmosphere to determine which genes ANAC102 is capable of regulating. Gene expression was also compared between three week old plants of an ANAC102 knock-out line and parental ecotype Col-0 under 0.1% Oxygen and ambient atmosphere conditions to determine which genes may require ANAC102 for appropriate expression under these conditions. Gene expression was also compared between imbibed seeds of an ANAC102 knock-out line and parental ecotype Col-0 following a 0.1% Oxygen treatment. Experiment Overall Design: Three related experiments were performed. In the first two, three week old Arabidopsis seedlings (4-6 leaves) were used. In the first experiment one line of ANAC102 over-expressing plants were compared to ecotype C24 (the parental line used in the creation of the ANAC102 over-expressing line). Two biological replicates were used for each line and for each biological replicate, five Arabidopsis seedlings grown on the same petri dish were bulked. RNA was extracted from whole plants. In the second experiment, one line of plants bearing a T-DNA insertion in the second exon of the ANAC102 gene was compared to ecotype Col-0 (the parental line used for the T-DNA mutagenesis) both without treatment and following treatment with 0.1% Oxygen for four hours. Two biological replicates were used for each line and for each biological replicate, five Arabidopsis seedlings grown on the same petri dish were bulked. RNA was extracted from root tissue only. In the third experiment imbibed seeds of one Arabiopsis line bearing a T-DNA insertion in the second exon of the ANAC102 gene was compared to seeds of ecotype Col-0 (the parental line used for the T-DNA mutagenesis) following treatment with 0.1% Oxygen for six days. Three biological replicates were used for each line and for each biological replicate, ~5mg (pre-imbibition) of seed were bulked. RNA was extracted from whole seed.

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Global gene expression analysis of Arabidopsis Col-0 under normal and low oxygen conditions

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