DRB1, DRB2 and DRB4 Are Required for an Appropriate miRNA-Mediated Molecular Response to Osmotic Stress in Arabidopsis thaliana

Joseph L. Pegler; Jackson M. J. Oultram; Christopher P. L. Grof; Andrew L. Eamens

doi:10.3390/ijms252312562

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DRB1, DRB2 and DRB4 Are Required for an Appropriate miRNA-Mediated Molecular Response to Osmotic Stress in Arabidopsis thaliana

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DRB1, DRB2 and DRB4 Are Required for an Appropriate miRNA-Mediated Molecular Response to Osmotic Stress in Arabidopsis thaliana

Joseph L. Pegler, Jackson M. J. Oultram, Christopher P. L. Grof and Andrew L. Eamens

International Journal of Molecular Sciences, Vol.25(23), pp.1-25

2024

DOI: https://doi.org/10.3390/ijms252312562

PMCID: PMC11641234

PMID: 39684274

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Abstract

Arabidopsis thaliana (Arabidopsis)

osmotic stress

mannitol

microRNA (miRNA)

miRNA-directed gene expression regulation

double-stranded RNA binding (DRB) protein

DRB1

DRB2

DRB4

Arabidopsis thaliana (Arabidopsis) double-stranded RNA binding (DRB) proteins DRB1, DRB2 and DRB4 perform essential roles in microRNA (miRNA) production, with many of the produced miRNAs mediating aspects of the molecular response of Arabidopsis to abiotic stress. Exposure of the drb1, drb2 and drb4 mutants to mannitol stress showed drb2 to be the most sensitive to this form of osmotic stress. Profiling of the miRNA landscapes of mannitol-stressed drb1, drb2 and drb4 seedlings via small RNA sequencing, and comparison of these to the profile of mannitol-stressed wild-type Arabidopsis plants, revealed that the ability of the drb1 and drb2 mutants to mount an appropriate miRNA-mediated molecular response to mannitol stress was defective. RT-qPCR was next used to further characterize seven miRNA/target gene expression modules, with this analysis identifying DRB1 as the primary DRB protein required for miR160, miR164, miR167 and miR396 production. In addition, via its antagonism of DRB1 function, DRB2 was shown by RT-qPCR to play a secondary role in regulating the production of these four miRNAs. This analysis further showed that DRB1, DRB2 and DRB4 are all required to regulate the production of miR399 and miR408, and that DRB4 is the primary DRB protein required to produce the non-conserved miRNA, miR858. Finally, RT-qPCR was used to reveal that each of the seven characterized miRNA/target gene expression modules responded differently to mannitol-induced osmotic stress in each of the four assessed Arabidopsis lines. In summary, this research has identified mannitol-stress-responsive miRNA/target gene expression modules that can be molecularly manipulated in the future to generate novel Arabidopsis lines with increased tolerance to this form of osmotic stress.

Details

Title: DRB1, DRB2 and DRB4 Are Required for an Appropriate miRNA-Mediated Molecular Response to Osmotic Stress in Arabidopsis thaliana
Authors: Joseph L. Pegler - University of Newcastle Australia
Jackson M. J. Oultram - University of Newcastle Australia
Christopher P. L. Grof - University of Newcastle Australia
Andrew L. Eamens (Corresponding Author) - University of the Sunshine Coast, Queensland, School of Health - Biomedicine
Publication details: International Journal of Molecular Sciences, Vol.25(23), pp.1-25
Publisher: MDPI AG
Date published: 2024
DOI: 10.3390/ijms252312562
ISSN: 1422-0067
PMID: 39684274; PMC11641234
Copyright note: © 2024 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Data Availability: All data reported here are available from the authors upon request.
Organisation Unit: School of Health - Biomedicine
Language: English
Record Identifier: 991082887102621
Output Type: Journal article

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