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The rice pseudo phosphotransfer proteins are negative regulators of endogenous cytokinin signaling and affect multiple aspects of growth and development.

ABSTRACT: The phytohormone cytokinin plays a significant role in nearly all aspects of plant growth and development. Cytokinin signaling has primarily been studied in the dicot model Arabidopsis, with relatively little work done in monocots, which include rice (Oryza sativa) and other cereals of agronomic importance. The cytokinin signaling pathway is a phosphorelay comprised of the histidine kinase receptors (HKs), the authentic histidine phosphotransfer proteins (AHPs), and the type-B Response Regulators (ARRs). Two negative regulators of cytokinin signaling have been identified: the type-A ARRs, which are cytokinin primary response genes, and the pseudo histidine phosphotransfer proteins (PHPs), which lack the His residue required for phosphorelay. Here, we describe the role of the PHP genes from rice. Phylogenic analysis indicates that the PHPs are generally first found in the genomes of gymnosperms and that they arose independently in monocots and dicots. Consistent with this, the three rice PHPs fail to complement an Arabidopsis php mutant (ahp6). Disruption of the three PHPs results in a molecular phenotype consistent with these elements acting as negative regulators of basal cytokinin signaling, including the constitutive up-regulation of a number of type-A RR and cytokinin oxidase genes. The triple php mutant affects multiple aspects of rice growth and development, including shoot morphology, panicle architecture and seed fill. However, in contrast to Arabidopsis, disruption of the rice PHPs does not affect root vascular patterning, suggesting that while many aspects of key signaling networks are conserved between monocots and dicots, the molecular components regulating some processes are distinct.

ORGANISM(S): Oryza sativa Japonica Group

PROVIDER: GSE141408 | GEO | 2022/12/31

REPOSITORIES: GEO

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Project description:In Arabidopsis thaliana, the immediate early response of plants to cytokinin is formulated as the multistep AHK-AHP-ARR phosphorelay signaling circuitry, which is initiated by the cytokinin-receptor histidine protein kinases. In the hope of finding components (or genes) that function downstream of the cytokinin-mediated His-Asp phosphorelay signaling circuitry, we carried out genome-wide microarray analyses. To this end, we focused on a pair of highly homologous ARR10 and ARR12 genes by constructing an arr10 arr12 double null mutant. The mutant alleles used in this study were arr10-5 and arr12-1. arr10-5 is the SALK_098604 T-DNA insertion line, whose mutation was determined to be located in the fifth exon of the ARR10 coding sequence. Arr12-1 is the SALK_054752 T-DNA insertion line, whose mutation was determined to be located in the third exon of the ARR12 coding sequence. The resulting mutant exhibits a characteristic phenotype with regard to the cytokinin-mediated His-Asp phosphorelay. Here we, therefore, compared response to cytokinin in wild type with that in arr10 arr12 double mutant. In this study, wild type and the arr10 arr12 double mutant grown vertically on MS agar plates for 2 weeks were treated with 20uM t-zeatin or 0.02% DMSO (solvent for t-zetion solution) for 1h. These treated plant samples were divided into three portions, from which RNA samples were prepared separately from aerial parts of seedlings with use of RNeasy Plant Mini Kit (Qiagen, Valencia, CA, U.S.A.). The Quality of RNAs prepared was analyzed by Bioanalyzer 2100 (Agilent Technologies). These RNA samples were processed as recommended by the Affymetrix instruction (Affymetrix GeneChip Expression Analysis Technical Manual, Affymetrix). These dataset will provide us with bases for understanding the early response to cytokinin on aerial parts of seedlings in Arabidopsis thaliana. Experimenter name: Hitoshi SAKAKIBARA; Experimenter phone: 81455039576; Experimenter fax: 81455039609; Experimenter address: Biodynamics Research Team; Experimenter address: RIKEN Plant Science Center; Experimenter address: 1-7-22 Suehiro, Tsurumi; Experimenter address: Yokohama; Experimenter zip/postal_code: 230-0045; Experimenter country: JAPAN Experiment Overall Design: 12 samples were used in this experiment

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The rice pseudo phosphotransfer proteins are negative regulators of endogenous cytokinin signaling and affect multiple aspects of growth and development.

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