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Plasma membrane proteomics in the maize primary root elongation zone under water stress conditions

ABSTRACT: Maintenance of root growth is critical to plant adaptation to drought conditions. Previous work on the maize (Zea mays L.) primary root under water stress showed that cell elongation is maintained in the apical region of the growth zone but progressively inhibited further from the apex. These responses involve spatially differential and coordinated regulation of cellular growth processes, including modifications of both cell production rate and cell wall extensibility. As the interface between the cytoplasm and the apoplast (including the cell wall), the plasma membrane is likely to play major functions in the coordination of cell production and expansion. In addition, plasma membrane proteins may be involved in solute uptake for osmotic adjustment, pH regulation, ion homeostasis and other critical processes in roots growing under water-stressed conditions. Due to technical limitations, however, plasma membrane proteomic studies have not been reported for water-stressed tissues. Using a simplified method for enrichment of plasma membrane proteins, we compared the developmental distribution of plasma membrane proteins that are differentially regulated in the growth zone of well-watered and water-stressed roots.

INSTRUMENT(S): LTQ Orbitrap

ORGANISM(S): Zea Mays (maize)

TISSUE(S): Plant Cell, Root

SUBMITTER: Jeff Anderson

LAB HEAD: Scott Peck

PROVIDER: PXD002027 | Pride | 2020-11-11

REPOSITORIES: Pride

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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).

			Action	DRS
	2_D1_1.RAW	Raw
	2_D1_10.RAW	Raw
	2_D1_11.RAW	Raw
	2_D1_12.RAW	Raw
	2_D1_13.RAW	Raw

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Plasma membrane proteomics in the maize primary root elongation zone under water stress conditions

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