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Protein Phosphorylation Orchestrates Acclimations of Arabidopsis Plants to Environmental pH

ABSTRACT: In lieu of strategies to escape unfavorable conditions, plants have evolved mechanisms to cope with and adapt to changes in their environment. Environment pH (pHe) is a key parameter that dictates a surfeit of conditions of critical importance for plant survival and fitness. Besides responses to altered availability of essential nutrients, changes in pHe recalibration of cytoplasmic and apoplastic pH despite constant fluctuations. To elucidate processes allowing such acclimations, we conducted a comprehensive proteomic/phosphoproteomic survey of plants subjected to transient exposure to acidic or alkaline pH, an approach that covered more than 60% of the protein-coding genes of the model plant Arabidopsis thaliana. Differentially (phospho)peptides displayed moderate concordance to previously published transcriptomic data sets, indicative of extensive post-transcriptomic regulation. This finding is corroborated by the discovery of xx so far undocumented, possibly pH-specific phospho-sites in response to alterations in pHe. Both low and high pHe altered proton-translocation across the plasma membrane. In pH 7.5 plants, transport but not assimilation of nitrogen was strongly induced, possibly to increase cytosolic H+ concentration. Mutants defective in the nitrate transporter nrt2.1 exhibited longer roots at low pH and increased density of root hairs at high pH, suggesting a moonlighting role of NRT2.1 in root growth. Only a few pH-responsive proteins were associated with the transport of iron and zinc, the availability of which is altered by pHe. Intricate phosphorylation of the ABC transporters PDR7 and PDR8 suggests pH-dependent phospho-switches in substrate specificity, tipping the balance between growth and defense. Unexpectedly, pHe provoked pronounced, changes in the leaf proteome, possibly communicated via an elaborate calcium-dependent signaling network. Moreover, we show that the TPLATE compounds EH1 and EH2 are differentially phosphorylated at multiple sites in response to pHe, indicating that the endocytic cargo protein trafficking is orchestrated by pHe.

INSTRUMENT(S): Orbitrap Fusion Lumos

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

TISSUE(S): Root, Shoot

SUBMITTER: Chuan-Chih Hsu

LAB HEAD: Wolfgang Schmidt

PROVIDER: PXD045226 | Pride | 2023-09-18

REPOSITORIES: Pride

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			Action	DRS
	A426_Root_total_Mascot.msf	Msf
	A426_Root_total_Mascot.msfView	Msf
	A426_Root_total_Mascot.pdResult	Other
	A426_Root_total_Mascot.pdResultView	Other
	A426_Root_total_TMT_Sequest.msf	Msf

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Project description:Azospirillum is a plant growth promoting rhizobacteria (PGPR) with ability to produce several phytohormones such as auxins, mainly indole-3-acetic acid (IAA). The positive interaction of Azospirillum with plants has been simplified and explained through the bacterial capacity to produce IAA. Typical changes on root architecture by promoting the number of lateral roots and hair formation, and reducing the primary root length were established in inoculated plants. These changes increase the root surface improving the water and nutrients acquisition, and thus the growth of the whole plant. The mechanisms by which Azospirillum induces such changes fails to be explained only by the bacterial capacity to produce IAA. In this work, we have evaluated the root architecture and gene expression changes occurred in Arabidopsis thaliana inoculated with A. brasilense Az39 and the IAA-deficient mutant (Az39 ipdC-), or treated with exogenous IAA solution to confirm both, the IAA-dependent and IAA-independent Azospirillum´s pathways to promote the root growth. Our results demonstrate the ability of Az39 to modify the primary root development through IAA biosynthesis, while other IAA-independent mechanisms were related to an increase in the lateral roots development and the root hairs number. Jasmonates, ethylene and salicylic acid were increased in the IAA-deficient bacterial treatments, as the ipdC mutant significantly up-regulated transcription of genes enriched of these phytohormones signaling after 7 days. Further, the physical presence of the inactive bacteria (Az39φ) seems to mediate the development of root hairs, a mechanism common to other non-PGPR as E. coli DH5α. Our results suggest that Az39 inoculation induces morphological changes in root architecture through both IAA-dependent and independent mechanism. The IAA biosynthesis by Az39 reduces the primary root length; while the cells contact with the roots increases the root hairs production. Both the synthesis of active IAA and the presence of metabolically active Az39 cells increase the growth and development of lateral roots.

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Protein Phosphorylation Orchestrates Acclimations of Arabidopsis Plants to Environmental pH

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