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Col-0_srk2de_abi1-1C_GCPs_Proteomics

ABSTRACT: Gas exchange between plants and the atmosphere is critical for regulating plant growth or stress responses. When plants are exposed to drought stress, abscisic acid (ABA)-activated SnRK2 protein kinases play a central role for stomatal closure to prevent excessive water loss. To identify SnRK2 substrates in guard cells, a cell-type-specific phosphoproteomic analysis was performed, and MAP4K1 and MAP4K2 were identified as putative signaling factors in guard cells. MAP4K1 and MAP4K2 function redundantly in guard cells, as a map4k1map4k2 mutant impaired ABA-induced stomatal closure. MAP4K1 Ser-479 is directly phosphorylated by SRK2E (OST1/SnRK2.6), and it physiologically interacts with SRK2E in planta, suggesting that MAP4K1 is in vivo substrates of SnRK2. Mutational analysis revealed that phosphorylation of MAP4K1 Ser-479 is required for ABA-induced stomatal closure. Furthermore, an electrophysiological assay suggested that MAP4K1/2 are involved in the ABA-responsive activation of Ca2+-permeable channels. Collectively, our results propose that ABA-activated SnRK2 and MAP4K1/2 constitute a signaling pathway that positively regulates ABA-induced stomatal closure via Ca2+ signaling.

ORGANISM(S): Arabidopsis Thaliana (mouse-ear Cress)

SUBMITTER: Kota Yamashita

PROVIDER: PXD049021 | JPOST Repository | Thu Jan 30 00:00:00 GMT 2025

REPOSITORIES: jPOST

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			Action	DRS
	test211204_Col-0_GCP_ABA00min_rep1_Proteome.raw	Raw
	test211204_Col-0_GCP_ABA02min_rep1_Proteome.raw	Raw
	test211204_Col-0_GCP_ABA10min_rep1_Proteome.raw	Raw
	test211204_Col-0_GCP_ABA30min_rep1_Proteome.raw	Raw
	test211204_abi1-1C_GCP_ABA00min_rep1_Proteome.raw	Raw

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Project description:Transgenic Arabidopsis plants with constitutively low inositol (1,4,5) triphosphate exhibit an increased tolerance to water stress by an ABA-independent pathway; The phosphoinositide pathway and inositol (1,4,5) trisphopsphate (InsP3) are implicated in plant responses to stress. In order to manipulate the pathway and determine the downstream consequences of altered InsP3-mediated signaling, we generated transgenic Arabidopsis plants expressing the mammalian type I inositol polyphosphate 5-phosphatase, an enzyme that specifically hydrolyzes the soluble inositol phosphates and terminates the signal. Transgenic plants have no morphological differences compared to wild type; however, rapid transient Ca2+ responses to a cold or salt stimulus are reduced by ~ 30%. To further understand the role of InsP3-mediated signaling in plant stress responses we focused on drought stress. Surprisingly, the InsP 5-ptase plants lose less water and exhibited an increased tolerance to drought. Stomatal bioassays showed that transgenic guard cells are less responsive to the inhibition of opening by ABA but show an increased sensitivity to ABA-induced closure. The onset of the drought stress is delayed in the transgenic plants and ABA levels did not increase as much as in the wild type. Transcript profiling has revealed that DREB2A and a subset of DREB2A regulated genes are basally up regulated in the InsP 5-ptase plants. These results indicate that the drought tolerance of the InsP 5-ptase plants is mediated in part via an ABA-independent pathway. The constitutive dampening of the InsP3 signal in this system has uncovered novel regulation and cross talk between signaling pathways. Experiment Overall Design: In order to compare global expression profiles between wild type and transgenic plants in response to water stress we harvested leaves for transcript profiling at day 0 (well watered) and at day 7 when the transgenic plants were still at an RWC of >85% and the wild type and vector control plants were at an RWC of ~ 50 -60% . Leaves were harvested and pooled from ~ 10 plants of each line/time point and three independent biological replicates were carried out with wild type (WT) and 2 independent transgenic lines (T6 and T8). The vector control line (C2) was used as an additional control.

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Col-0_srk2de_abi1-1C_GCPs_Proteomics

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