Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Comparison of low water potential (drought)-regulated gene expression in wild type (Col-0) and the hai1-2 (At5g59220) mutant

ABSTRACT: The Clade A PP2C Highly ABA-Induced1 (HAI1, At5g59220) is strongly up-regulated by low water potential in an ABA-dependent manner. Using knockout mutants of hai1, we found that HAI1 functions as a negative regulator of low water potential-induced proline and osmoregulatory solute accumulation. We also found a relatively weak and limited interaction of HAI1 with the RCAR/PYL family of ABA receptors. This, plus its induced expression, suggest that HAI1 remains active during stress and attenuates specific aspects of drought response. Microarray experiments were used to identify genes differentially expressed in hai1-2 (SALK_108282) versus wild type under both unstress control conditions and after low water potential (i.e., drought, water deficit). A relatively long term (96 h) low water potential treatment was used as phenotypes of hai1-2 were most apparent after this longer term low water potential treatment. Seven-day-old seedlings of wild type (Col-0) or hai1-2 were transferred from agar plates of control media to low water potential stress (PEG-infused agar plates at -1.2 MPa) or to fresh control media (-0.25 Mpa). Samples (whole seedlings) were collected 96 h after transfer. Three independent experiments were conducted with one sample of each treatment (wild type stress and control, hai1-2 stress and control) collected from each experiment.

ORGANISM(S): Arabidopsis thaliana

SUBMITTER: Nguyen Thao

PROVIDER: E-GEOD-35258 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Unique drought resistance functions of the highly ABA-induced clade A protein phosphatase 2Cs.

Bhaskara Govinal Badiger GB Nguyen Thao Thi TT Verslues Paul E PE

Plant physiology 20120724 1

Six Arabidopsis (Arabidopsis thaliana) clade A protein phosphatase 2Cs (PP2Cs) have established abscisic acid (ABA) signaling roles; however, phenotypic roles of the remaining three "HAI" PP2Cs, Highly ABA-Induced1 (HAI1), AKT1-Interacting PP2C1/HAI2, and HAI3, have remained unclear. HAI PP2C mutants had enhanced proline and osmoregulatory solute accumulation at low water potential, while mutants of other clade A PP2Cs had no or lesser effect on these drought resistance traits. hai1-2 also had i ...[more]

PMID: 22829320

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Project description:Previous work showed that the maize primary root adapts to low water potential (-1.6 MPa) by maintaining longitudinal expansion in the apical 3 mm (region 1), whereas in the adjacent 4 mm (region 2) longitudinal expansion reaches a maximum in well-watered roots but is progressively inhibited at low water potential. To identify mechanisms that determine these responses to low water potential, transcript expression was profiled in these regions of water-stressed and well-watered roots. In addition, comparison between region 2 of water-stressed roots and the zone of growth deceleration in well-watered roots (region 3) distinguished stress-responsive genes in region 2 from those involved in cell maturation. Responses of gene expression to water stress in regions 1 and 2 were largely distinct. The largest functional categories of differentially expressed transcripts were reactive oxygen species and carbon metabolism in region 1, and membrane transport in region 2. Transcripts controlling sucrose hydrolysis distinguished well-watered and water-stressed states (invertase vs. sucrose synthase), and changes in expression of transcripts for starch synthesis indicated further alteration in carbon metabolism under water deficit. A role for inositols in the stress response was suggested, as was control of proline metabolism. Increased expression of transcripts for wall-loosening proteins in region 1, and for elements of ABA and ethylene signaling were also indicated in the response to water deficit. The analysis indicates that fundamentally different signaling and metabolic response mechanisms are involved in the response to water stress in different regions of the maize primary root elongation zone. Keywords: water stress response, root growth Three pair-wise comparisons were made of transcripts from water-stressed and well-watered tissues in the different root tip regions. In the first comparison (C1), transcripts from region 1 of water-stressed seedlings were compared with those from region 1 of well-watered seedlings. The second comparison (C2) was made between transcripts from region 2 of the two treatments. We expected a larger number of genes to be differentially expressed in region 2 because its elongation rate decreased greatly under water-stressed compared with well-watered conditions. To prioritize the differentially expressed genes revealed in this comparison, a distinction was made between those genes that are associated with growth inhibition in region 2 specifically as a response to water stress, and those genes that are involved in root cell maturation whether under stress or control conditions. A hypothetical example of the former might be genes involved in auxin response since water stress can increase maize root auxin content [8] and application of exogenous auxin can shorten the root growth zone [9]. An example of the latter might involve genes for secondary wall synthesis, e.g., [10]. To experimentally make this distinction a third pair-wise comparison (C2/3) was included to compare expression of genes between water-stressed region 2 and well-watered region 3 as these are both regions of cell maturation. Genes differentially expressed in both C2 and C2/3 are more likely to cause growth inhibition at low water potential and are not likely to be part of the maturation program itself, whereas genes differentially expressed only in C2 are more likely related to maturation. Four biological replicates were included each with dye-swap. The maize oligonucleotide array comes in two slides, A and B. Thus for each of the three comparisons made there were 16 slides (4 biological reps, 2 for dye-swap, 2 parts to an array).

Dataset Information

Comparison of low water potential (drought)-regulated gene expression in wild type (Col-0) and the hai1-2 (At5g59220) mutant

Publications

Unique drought resistance functions of the highly ABA-induced clade A protein phosphatase 2Cs.

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