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Bulk RNA-sequencing of hypocotyl explants and hypocotyl explant-derived calli of Arabidopsis

ABSTRACT: Plants are aerobic organisms that rely on molecular oxygen for respiratory energy production. Hypoxic conditions, with oxygen levels ranging between 1% and 5%, usually limit aerobic respiration and affect plant growth and development. Here, we demonstrate that hypoxic microenvironment induced by active cell proliferation during the two-step plant regeneration process intrinsically represses the regeneration competence of callus in Arabidopsis thaliana. Hypoxia-repressed plant regeneration was mediated by the RELATED TO APETALA 2.12 (RAP2.12) protein, a member of the Ethylene Response Factor VII (ERF-VII) family. The hypoxia-activated RAP2.12 protein promoted salicylic acid (SA) biosynthesis and defense responses, inhibiting pluripotency acquisition and de novo shoot regeneration in calli. RAP2.12 could bind directly to the SALICYLIC ACID INDUCTION DEFICIENT 2 (SID2) gene promoter and activate SA biosynthesis, repressing plant regeneration via a PLETHORA (PLT)-dependent pathway. The rap2.12 mutant calli exhibited enhanced shoot regeneration, which was impaired by SA treatment. Taken together, our findings demonstrate that cell proliferation-dependent hypoxic microenvironment reduces cellular pluripotency and plant regeneration through the RAP2.12–SID2 module.

ORGANISM(S): Arabidopsis thaliana

PROVIDER: GSE249446 | GEO | 2024/02/16

REPOSITORIES: GEO

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Bulk RNA-sequencing of hypocotyl explant-derived calli of Arabidopsis WT and rap2.12-2

Project description:Plants are aerobic organisms that rely on molecular oxygen for respiratory energy production. Hypoxic conditions, with oxygen levels ranging between 1% and 5%, usually limit aerobic respiration and affect plant growth and development. Here, we demonstrate that hypoxic microenvironment induced by active cell proliferation during the two-step plant regeneration process intrinsically represses the regeneration competence of callus in Arabidopsis thaliana. Hypoxia-repressed plant regeneration was mediated by the RELATED TO APETALA 2.12 (RAP2.12) protein, a member of the Ethylene Response Factor VII (ERF-VII) family. The hypoxia-activated RAP2.12 protein promoted salicylic acid (SA) biosynthesis and defense responses, inhibiting pluripotency acquisition and de novo shoot regeneration in calli. RAP2.12 could bind directly to the SALICYLIC ACID INDUCTION DEFICIENT 2 (SID2) gene promoter and activate SA biosynthesis, repressing plant regeneration via a PLETHORA (PLT)-dependent pathway. The rap2.12 mutant calli exhibited enhanced shoot regeneration, which was impaired by SA treatment. Taken together, our findings demonstrate that cell proliferation-dependent hypoxic microenvironment reduces cellular pluripotency and plant regeneration through the RAP2.12–SID2 module.

2024-02-16 | GSE249447 | GEO

Expression data from WT, sid2-1

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Regulation of the microsomal proteome by salicylic acid and deficiency of phosphatidylinositol‐4‐kinases β1 and β2 in Arabidopsis thaliana

Project description:Phosphatidylinositol-4-kinases β1 and β2 (PI4Kβ1/PI4Kβ2), which are responsible for phosphorylation of phosphoinositol to phosphatidylinositol-4-phosphate, have an important role in vesicular trafficking in plants. Moreover, PI4Kβ1/PI4β2 negatively regulate biosynthesis of phytohormone salicylic acid (SA). SA plays crucial role in plant immune responses and its endogenous concentration strongly affects plant transcriptome. In this study we were focused on the effect of the PI4Kβ1/PI4Kβ2 and SA on the abundance of membrane proteins. For this purpose, we performed proteomic analysis on isolated microsomal fractions from leaves of four Arabidopsis thaliana genotypes: wild type ecotype Columbia-0; double mutant impaired in function of PI4Kβ1/PI4Kβ2 (pi4kβ1/pi4kβ2) exhibiting high SA level; sid2 mutant with impaired SA biosynthesis depending on the isochorismate synthase 1 and triple mutant sid2/pi4kβ1/pi4kβ2. In total, we identified 4534 proteins from which 1696 proteins differed in abundance between the mutants and WT. We showed that SA has a big impact on membrane proteome, because among hundreds of the identified affected proteins typical proteins associated with SA triggered pathway occurred. Our data thus point out new connections between SA pathway and clathrin independent endocytosis (flotillins) and exocytosis/protein secretion (syntaxins, tertraspanin) to be investigated in future. In contrast to SA, presence/absence of PI4Kβ1/PI4Kβ2 itself affected only 27 proteins. Nevertheless, among them we identified CERK1, plasma membrane receptor for chitin. Although PI4Kβ1/PI4Kβ2 itself did not have strong impact on A. thaliana microsomal proteome, our data clearly shows that PI4Kβ1/PI4Kβ2 enhance the proteome changes when SA pathway is modulated in parallel

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Expression data from WT, sid2-1

2013-06-30 | E-GEOD-43819 | biostudies-arrayexpress

Hypoxic gene expression in med25-1

Project description:Plants as obligate aerobic organisms are severely affected by flooding-induced oxygen restriction. As a consequence, they suffer from energy deficits which are counterbalanced by activation of anaerobic metabolism genes through subgroup VII ETHYLENE-RESPONSE FACTOR (ERFVII) transcription factors. Despite the crucial role of ERFVIIs in regulating transcript responses to hypoxia in multiple plant species, it is not understood how these factors mechanistically initiate hypoxic gene expression. Here, we show that the Arabidopsis ERFVIIs RELATED TO APETALA 2.12 (RAP2.12) and RAP2.2 recruit the Mediator complex, present in all higher eukaryotes, in order to convey stress signals to RNA polymerase II. This is achieved by specific interaction with Mediator subunit 25 (AtMED25). Both RAP2.12 and RAP2.2 require AtMED25 for full activity and subsequent induction of target hypoxia core genes. Our findings support an interplay of multiple coactivators under hypoxia and underline the flexible nature of the Mediator complex composition, which both help to meet specific cellular demands under (hypoxic) stress conditions.

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A Tripartite Amplification Loop Involving WRKY75, Salicylic Acid and ROS Accelerates Leaf Senescence in Arabidopsis

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RNAseq samples of CNQX treated Arabidopsis roots regenerating at different time points.

Project description:Wounding is a trigger for both regeneration and defense in plants, but it is not clear if the two responses are linked by common activation or represent potential tradeoffs. While plant glutamate-like receptors (GLRs) are known to mediate defense responses, we implicate GLRs in regeneration through dynamic changes in chromatin and transcription in reprogramming cells near wound sites. Here, we show that genetic mutation and pharmacological inhibition of GLRs increases regeneration efficiency in multiple organ repair systems and plant species. Perturbation of GLR-mediated function, possibly by affecting Calcium fluxes, speeds cell division and re-specification of the stem cell niche while dampening a subset of defense responses. We show that the GLRs work through salicylic acid (SA) signaling in their effects on regeneration, with mutants in the SA receptor NPR1 partially resistant to GLR perturbation and hyper regenerative. These findings reveal a conserved mechanism that regulates a tradeoff between defense and regeneration and also offer new strategies to improve regeneration in agricultural and conservation.

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