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PMID: 19889877 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The nuclear interactor PYL8/RCAR3 of Fagus sylvatica FsPP2C1 is a positive regulator of abscisic acid signaling in seeds and stress.

Plant physiology ·Vol. 152 ·No. 1 ·2010-01-00 ·Pages 133-50

Saavedra X, Modrego A, Rodríguez D, González-García MP, Sanz L, Nicolás G, Lorenzo O

Abstract

The functional protein phosphatase type 2C from beechnut (Fagus sylvatica; FsPP2C1) was a negative regulator of abscisic acid (ABA) signaling in seeds. In this report, to get deeper insight on FsPP2C1 function, we aim to identify PP2C-interacting partners. Two closely related members (PYL8/RCAR3 and PYL7/RCAR2) of the Arabidopsis (Arabidopsis thaliana) BetV I family were shown to bind FsPP2C1 in a yeast two-hybrid screening and in an ABA-independent manner. By transient expression of FsPP2C1 and PYL8/RCAR3 in epidermal onion (Allium cepa) cells and agroinfiltration in tobacco (Nicotiana benthamiana) as green fluorescent protein fusion proteins, we obtained evidence supporting the subcellular localization of both proteins mainly in the nucleus and in both the cytosol and the nucleus, respectively. The in planta interaction of both proteins in tobacco cells by bimolecular fluorescence complementation assays resulted in a specific nuclear colocalization of this interaction. Constitutive overexpression of PYL8/RCAR3 confers ABA hypersensitivity in Arabidopsis seeds and, consequently, an enhanced degree of seed dormancy. Additionally, transgenic 35S:PYL8/RCAR3 plants are unable to germinate under low concentrations of mannitol, NaCl, or paclobutrazol, which are not inhibiting conditions to the wild type. In vegetative tissues, Arabidopsis PYL8/RCAR3 transgenic plants show ABA-resistant drought response and a strong inhibition of early root growth. These phenotypes are strengthened at the molecular level with the enhanced induction of several ABA response genes. Both seed and vegetative phenotypes of Arabidopsis 35S:PYL8/RCAR3 plants are opposite those of 35S:FsPP2C1 plants. Finally, double transgenic plants confirm the role of PYL8/RCAR3 by antagonizing FsPP2C1 function and demonstrating that PYL8/RCAR3 positively regulates ABA signaling during germination and abiotic stress responses.

MeSH Terms
Abscisic Acid/metabolism Amino Acid Sequence Fagus/genetics,metabolism Gene Expression Regulation, Plant Intracellular Signaling Peptides and Proteins/genetics,metabolism Nuclear Proteins/genetics,metabolism Phylogeny Plant Leaves/cytology,metabolism Plant Proteins/genetics,metabolism Plants, Genetically Modified Seeds/metabolism Signal Transduction/physiology Stress, Physiological Tobacco Two-Hybrid System Techniques
Chemicals
Intracellular Signaling Peptides and Proteins Nuclear Proteins Plant Proteins Abscisic Acid
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Saavedra Xandra
Departamento de Fisiología Vegetal, Centro Hispano-Luso de Investigaciones Agrarias, Facultad de Biología, Universidad de Salamanca, 37185 Salamanca, Spain.
Modrego Abelardo
Rodríguez Dolores
González-García Mary Paz
Sanz Luis
Nicolás Gregorio
Lorenzo Oscar
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Article Info
Journal
Plant physiology
Abbr.
Plant Physiol
ISSN
1532-2548
DOI
10.1104/pp.109.146381
Published
2010-01-00
Epub
2009-00-04
Pages
133-50
Language
English
Region
United States
NLM ID
0401224
PMCID
PMC2799352
Subset
IM
External Links
PubMed source DOI: 10.1104/pp.109.146381 PMC Full Text View journal details
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