Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Microarray analysis of humic acid extract effects on Triticum aestivum growth

ABSTRACT: Humic substances are principal components of soil organic matter. They have ecological importance as they intervene in regulating a large number of chemical and biological processes that occur in natural ecosystems. Their ability to improve plant growth has been well established in diverse plant species and growth conditions, although the mechanism responsible for this biological action is poorly understood. Microarray analysis might give us more information about up or down regulation of different biological processes. Wheat plants have been grown hydroponically and treated with Humic acid. Seeds were germinated in obscurity during 10 days, and grown in nutrient solution during 10 days. Harvests were conducted 24 hours, 72 hours and 30 days after treatment application, in order to study early response or a more sustained effect during time.

ORGANISM(S): Triticum aestivum

SUBMITTER: Maria Garnica

PROVIDER: E-MTAB-11280 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Plant cells contain different O-acetylserine(thiol)lyase (OASTL) enzymes involved in Cys biosynthesis and located in different subcellular compartments. These enzymes are made up of a complex variety of isoforms resulting in different subcellular Cys pools. To unravel the contribution of cytosolic Cys to plant metabolism, we characterized the knockout oas-a1.1 and osa-a1.2 mutants, deficient in the most abundant cytosolic OASTL isoform in Arabidposis thaliana. Total intracellular Cys and glutathione concentrations were reduced, and the glutathione redox state was shifted in favour of its oxidized form. Interestingly, the capability of the mutants to chelate heavy metals did not differ from that of the wild type, but the mutants have an enhanced sensitivity to Cd. With the aim of establishing the metabolic network most influenced by the cytosolic Cys pool, we used the ATH1 GeneChip for evaluation of differentially expressed genes in the oas-a1.1 mutant grown under non-stress conditions. The transcriptomic footprints of mutant plants had predicted functions associated with various physiological responses that are dependent on reactive oxygen species and suggested that the mutant was oxidatively stressed. To further elucidate the specific function(s) of the OAS-A1 isoform in the adaptation response to cadmium we extended the trasncriptome experiment to the wild type and oas-a1.1 mutant plants exposed to Cd. The comparison of transcriptomic profiles showed a higher proportion of genes with altered expression in the mutant than in the wild type, highlighting up-regulated genes identified as of the general oxidative stress response rather than metal-responsive genes. Wild type and oas-a1.1 mutant plants were grown hydroponically and, after a two-week acclimation period, the roots and shoots were harvested separately. Total RNA was then prepared and analyzed using the Affymetrix-Arabidopsis ATH1GeneChip array. Three biological replicates were performed for each sample. We made two different comparisons to classify the differently expressed genes in the mutant plant: oas-a1.1 roots versus wild-type roots and oas-a1.1 shoots versus wild-type shoots. Hydroponically-grown wild type and oas-a1.1 mutant plants were further treated with 50µM CdCl2 and 18h-treated-roots and 24h-treated-shoots were harvested. Total RNA was then prepared and analyzed using the Affymetrix-Arabidopsis ATH1GeneChip array. Three biological replicates were performed for each sample. Different comparisons were performed as follows: 18h Cd-treated wild type roots versus untreated wild type roots; 24h Cd-treated wild type shoots versus untreated wild type shoots; 18h Cd-treated oas-a1.1 roots versus untreated oas-a1.1 roots; 24h Cd-treated oas-a1.1 shoots versus untreated oas-a1.1 shoots; 18h Cd-treated oas-a1.1 roots versus 18h Cd-treated wild type roots; 24h Cd-treated oas-a1.1 shoots versus 24h Cd-treated wild type shoots

Project description:Background: Solubilized aluminum (Al) is rhizotoxic and can be present in acid soils at levels that inhibit root growth. Acid soils are found throughout the world and often support forests. However, unlike in herbaceous plants, little is known about the mechanisms by which forest trees respond to and tolerate Al. To begin to elucidate these mechanisms, we characterized transcriptomic changes in response to Al in roots of aspen (Populus tremula L.). Results: Aspen roots were treated with Al in solution culture for 6 h, 54 h, and 246 h. Transcriptomic changes were assessed by the Affymetrix GeneChip Poplar Genome Array. The analysis revealed 176 induced and 66 suppressed genes. The majority of these genes were regulated at 6 h, presumably reflecting root growth, which was strongly inhibited at 6 h and partially was recovered at 2 d and 10 d. Enrichment analysis identified sets of functionally related genes whose members were statistically over-represented compared to the genes on the microarray. These sets included genes related to cell wall modification, oxidative stress, cell death, and transport processes. Two of the genes involved in transport were related to the Arabidopsis Al tolerance genes AtALS3 (aluminum sensitive 3), possibly mediating redistribution of Al, and AtMATE (multi-drug and toxin extrusion), facilitating exudation of citrate. Expression patterns in response to Al in different plant tissues indicated that the two aspen genes are homologs of AtALS3 and AtMATE, suggesting that aspen and Arabidopsis share common mechanisms to cope with Al. Conclusion: This is the first survey of transcriptomic changes in response to Al in a forest tree. The results of our study provide a valuable set of data, which will help to further our understanding of the mechanisms and their regulation that enable aspen to grow in environments with toxic levels of Al. Aspen roots were treated with Al in solution culture for 6 h, 54 h, 246 h to cover a broad range of exposure times. Transcriptomic changes were assessed by the Affymetrix GeneChip Poplar Genome Array. For each time point, root tips of three independant plants were analyzed.

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Microarray analysis of humic acid extract effects on Triticum aestivum growth

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