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Gibberellin (GA) Treatment of Arabidopsis Root Tips

ABSTRACT: The plant hormone gibberellin (GA) represents an important regulator of growth and development. Early transcriptional events controlled by GA are not well characterised. Previous microarray studies have identified genes responsive to GA treatment in the whole seedling. The whole seedling represents many tissues where subtle effects of GA treatment in specific tissues may be masked. When treated with GA, an effect on the growth rate of roots was observed. More specifically, the shorter root of a GA-deficient plant can be rescued to wild-type length by the application of GA. This experiment was designed to identify GA-regulated genes in the root tips of Arabidopsis. The use of a GA-deficient mutant provides a greater potential to identify genes responding to GA treatment. Root tips are ideally suited for the quick uptake of the hormone treatment. There will be two biological replicates which will each consist of a control treatment at 0 minutes and 2 hours, as well as the experimental GA-treated 2 hour time point. This system provides an opportunity to compare gene expression between treated and non-treated root tips and allow the identification of early GA-responsive genes.

ORGANISM(S): Arabidopsis thaliana

PROVIDER: GSE20223 | GEO | 2010/04/20

SECONDARY ACCESSION(S): PRJNA124153

REPOSITORIES: GEO

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Project description:Two winter wheat cultivars possessing a high tolerance to Al (Triticum aestivum L. cv. Atlas66 and the near isogenic line OK91G106, named Century-T in this work [59]) and two winter wheat cultivars with low tolerance to Al (T. aestivum L. cv. Bounty and the near isogenic line OK91G108, named Century-S in this work [59]) were grown as previously described, and treated under conditions where Al remains mostly under the Al3+ form [10]. To reduce pH variations and ensure that Al speciation was stable throughout the experiment, at least 100 ml of solution was used for each plant. The root growth inhibition (RGI) is expressed as 100 × [1- (root growth of Al-treated seedling divided by the root growth of control seedlings)]. Root tips (5-10 mm) were isolated after 24 hours of exposure to Al and frozen on dry ice. Total RNA was isolated from the root tips (5 mm long) of 50 plants collected from the various genotypes using the RNeasy Plant Mini Kit (Qiagen). The RNA quality was assessed on agarose gels and with the Bioanalyzer 2100 (Agilent). Microarray profiling was performed according to Affymetrix protocols at the Functional Genomics Platform of the McGill University and Génome Québec Innovation Centre using the Affymetrix GeneChip Wheat Genome Array. Plants of the different lines were treated with Al concentrations resulting in 50% RGI: 50 ?M for the tolerant cultivars Atlas66 and Century-T and 5 ?M for the sensitive cultivars Bounty and Century-S. Three biological replicates were used and the results were analyzed using the Robust Multiarray Analysis (RMA version 0.2) freeware. Atlas66 and Bounty plants not exposed to Al were used as control for all the other conditions. Genes showing a differential expression greater than two-fold (RMA differential expression of log2 ? 1) between the Atlas66 and Bounty controls were excluded from the reference set as these may represent cultivar dependent differences in basal expression levels that could bias the analysis. Overall, this excluded 1.26% of the genes on the microarray (773 of 55,052) demonstrating that most genes are expressed at a similar level between the two cultivars. The microarray headings are the following : For each array headings: The first term represents the cultivar: A = Atlas66; B = Bounty, 106 = Century-T, 108 = Century-S. The second twerm represents the Al concentration: 0 = no aluminum 5 = 5 ?M aluminum and 50 = 50 ?M aluminum. The last term represents the sample replicate number S1, S2 or S3.

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Gibberellin (GA) Treatment of Arabidopsis Root Tips

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