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      Meta‐analysis and meta‐regression of transcriptomic responses to water stress in Arabidopsis

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          Summary

          The large amounts of transcriptome data available for Arabidopsis thaliana make a compelling case for the need to generalize results across studies and extract the most robust and meaningful information possible from them. The results of various studies seeking to identify water stress‐responsive genes only partially overlap. The aim of this work was to combine transcriptomic studies in a systematic way that identifies commonalities in response, taking into account variation among studies due to batch effects as well as sampling variation, while also identifying the effect of study‐specific variables, such as the method of applying water stress, and the part of the plant the mRNA was extracted from. We used meta‐analysis, the quantitative synthesis of independent research results, to summarize expression responses to water stress across studies, and meta‐regression to model the contribution of covariates that may affect gene expression. We found that some genes with small but consistent differential responses become evident only when results are synthesized across experiments, and are missed in individual studies. We also identified genes with expression responses that are attributable to use of different plant parts and alternative methods for inducing water stress. Our results indicate that meta‐analysis and meta‐regression provide a powerful approach for identifying a robust gene set that is less sensitive to idiosyncratic results and for quantifying study characteristics that result in contrasting gene expression responses across studies. Combining meta‐analysis with individual analyses may contribute to a richer understanding of the biology of water stress responses, and may prove valuable in other gene expression studies.

          Significance Statement

          Transcriptome studies are sometimes difficult to compare due to different experimental conditions. Here we show that that meta‐analysis and meta‐regression are useful for identifying robust gene sets and can quantify study characteristics that result in contrasting gene expression responses across studies. Combining meta‐analysis with individual analyses thus contribute to a richer understanding of the biology of water stress responses and may prove valuable in other gene expression studies.

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          Most cited references30

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          Monitoring the expression profiles of 7000 Arabidopsis genes under drought, cold and high-salinity stresses using a full-length cDNA microarray.

          Full-length cDNAs are essential for functional analysis of plant genes in the post-sequencing era of the Arabidopsis genome. Recently, cDNA microarray analysis has been developed for quantitative analysis of global and simultaneous analysis of expression profiles. We have prepared a full-length cDNA microarray containing approximately 7000 independent, full-length cDNA groups to analyse the expression profiles of genes under drought, cold (low temperature) and high-salinity stress conditions over time. The transcripts of 53, 277 and 194 genes increased after cold, drought and high-salinity treatments, respectively, more than fivefold compared with the control genes. We also identified many highly drought-, cold- or high-salinity- stress-inducible genes. However, we observed strong relationships in the expression of these stress-responsive genes based on Venn diagram analysis, and found 22 stress-inducible genes that responded to all three stresses. Several gene groups showing different expression profiles were identified by analysis of their expression patterns during stress-responsive gene induction. The cold-inducible genes were classified into at least two gene groups from their expression profiles. DREB1A was included in a group whose expression peaked at 2 h after cold treatment. Among the drought, cold or high-salinity stress-inducible genes identified, we found 40 transcription factor genes (corresponding to approximately 11% of all stress-inducible genes identified), suggesting that various transcriptional regulatory mechanisms function in the drought, cold or high-salinity stress signal transduction pathways.
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            ABA-mediated transcriptional regulation in response to osmotic stress in plants.

            The plant hormone abscisic acid (ABA) plays a pivotal role in a variety of developmental processes and adaptive stress responses to environmental stimuli in plants. Cellular dehydration during the seed maturation and vegetative growth stages induces an increase in endogenous ABA levels, which control many dehydration-responsive genes. In Arabidopsis plants, ABA regulates nearly 10% of the protein-coding genes, a much higher percentage than other plant hormones. Expression of the genes is mainly regulated by two different families of bZIP transcription factors (TFs), ABI5 in the seeds and AREB/ABFs in the vegetative stage, in an ABA-responsive-element (ABRE) dependent manner. The SnRK2-AREB/ABF pathway governs the majority of ABA-mediated ABRE-dependent gene expression in response to osmotic stress during the vegetative stage. In addition to osmotic stress, the circadian clock and light conditions also appear to participate in the regulation of ABA-mediated gene expression, likely conferring versatile tolerance and repressing growth under stress conditions. Moreover, various other TFs belonging to several classes, including AP2/ERF, MYB, NAC, and HD-ZF, have been reported to engage in ABA-mediated gene expression. This review mainly focuses on the transcriptional regulation of ABA-mediated gene expression in response to osmotic stress during the vegetative growth stage in Arabidopsis.
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              RankProd: a bioconductor package for detecting differentially expressed genes in meta-analysis.

              While meta-analysis provides a powerful tool for analyzing microarray experiments by combining data from multiple studies, it presents unique computational challenges. The Bioconductor package RankProd provides a new and intuitive tool for this purpose in detecting differentially expressed genes under two experimental conditions. The package modifies and extends the rank product method proposed by Breitling et al., [(2004) FEBS Lett., 573, 83-92] to integrate multiple microarray studies from different laboratories and/or platforms. It offers several advantages over t-test based methods and accepts pre-processed expression datasets produced from a wide variety of platforms. The significance of the detection is assessed by a non-parametric permutation test, and the associated P-value and false discovery rate (FDR) are included in the output alongside the genes that are detected by user-defined criteria. A visualization plot is provided to view actual expression levels for each gene with estimated significance measurements. RankProd is available at Bioconductor http://www.bioconductor.org. A web-based interface will soon be available at http://cactus.salk.edu/RankProd
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                Author and article information

                Contributors
                Joshua.Rest@stonybrook.edu
                Jessica.Gurevitch@stonybrook.edu
                Journal
                Plant J
                Plant J
                10.1111/(ISSN)1365-313X
                TPJ
                The Plant Journal
                John Wiley and Sons Inc. (Hoboken )
                0960-7412
                1365-313X
                February 2016
                12 February 2016
                : 85
                : 4 ( doiID: 10.1111/tpj.2016.85.issue-4 )
                : 548-560
                Affiliations
                [ 1 ] Department of Ecology and Evolution, 650 Life SciencesStony Brook University Stony Brook NY 11794–5245USA
                [ 2 ] Center for Genomics and Systems BiologyNew York University 12 Waverly Place New York NY 10003USA
                Author notes
                Article
                TPJ13124
                10.1111/tpj.13124
                4815425
                26756945
                b25eb872-54b6-4ed0-9527-271609b1913f
                © 2016 The Authors. The Plant Journal published by Society for Experimental Biology and John Wiley & Sons Ltd.

                This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

                History
                : 01 December 2014
                : 04 January 2016
                : 08 January 2016
                Page count
                Figures: 4, Tables: 2, Pages: 13, Words: 10809
                Funding
                Funded by: US National Science Foundation Plant Genome Research
                Award ID: IOS‐1126971
                Funded by: US National Science Foundation Advances in Bioinformatics
                Award ID: NSF 1262402
                Funded by: Stony Brook University
                Funded by: US National Institutes of Health/National Institute of General Medical Sciences
                Award ID: R01 GM108904‐02
                Categories
                Technical Advance
                Technical Advance
                Custom metadata
                2.0
                tpj13124
                February 2016
                Converter:WILEY_ML3GV2_TO_NLMPMC version:4.9.8 mode:remove_FC converted:01.12.2016

                Plant science & Botany
                arabidopsis thaliana,drought response,water stress,meta‐analysis,research synthesis,gene expression microarray,transcriptomics

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