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Expression data from stratified anac089-2 mutant and wild-type seeds, ABA-treated.

ABSTRACT: Through a genetic screening in 3μM (+)-S-ABA, we have isolated the anac089-2 mutant, which was obtained from fast neutron-mutagenized Col-0 M2 seeds. This mutant showed ABA-insensitive phenotypes in the transition from dormancy to germination. anac089-2 mutant line was subjected to mapping based on Col/Ler polymorphisms. This mutation was independently mapped to the top arm of Chromosome 5. The anac089-2 mutation is located in a 25 kb region between CAPS markers MWD9/EcoRV and MWD9-25.3/AccI. This 25 kb region contains 8 genes, and sequencing of these genes revealed that anac089-2 mutant contained a mutation in the At5g22290. This gene encodes ANAC089, a nuclear localized NAC transcription factor, with the deduced protein of 340 amino acids. Southern blot analysis suggests that anac089-2 mutation is a ~700 bp deletion that includes the C-terminal region. ANAC089 is involved in an ABA signaling pathway during seed germination and stress responses (Osuna et al., 2011).

ORGANISM(S): Arabidopsis thaliana

PROVIDER: GSE25159 | GEO | 2019/12/31

REPOSITORIES: GEO

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Project description:Abscisic acid (ABA) is a key hormone for plant growth, development and stress adaptation. Perception of ABA through four types of receptors has been reported. We show here that impairment of ABA perception through the PYR/PYL/RCAR branch reduces vegetative growth and seed production, and leads to a severely open stomata and dramatic ABA insensitive phenotype, even though other branches for ABA perception remain functional. Arabidopsis sextuple mutant impaired in 6 PYR/PYL receptors, namely PYR1, PYL1, PYL2, PYL4, PYL5 and PYL8, was able to germinate and grow even on 100 mM ABA. Whole-rosette stomatal conductance (Gst) measurements revealed that leaf transpiration in the sextuple pyr/pyl mutant was higher than in the ABA-deficient aba3-1 or ABA-insensitive snrk2.6 mutants. The gradually increasing Gst values of plants lacking three, four, five and six PYR/PYLs indicate quantitative regulation of stomatal aperture by this family of receptors. The sextuple mutant lacked ABA-mediated activation of SnRK2s and ABA-responsive gene expression was dramatically impaired as was reported in snrk2.2/2.3/2.6. In summary, these results show that ABA perception by PYR/PYLs plays a major role to regulate seed germination and establishment, basal ABA signaling required for vegetative and reproductive growth, stomatal aperture and transcriptional response to the hormone. Four biological replicates were generated for the 3 genotypes, pyr1pyl1pyl2pyl4pyl5pyl8 sextuple mutant, snrk2.2/2.3/2.6 triple mutant, and Col-0. All the samples were treated with ABA for 3 hours before harvesting. The four Col-0 samples were mix to create an unique reference Col-0 sample. Two comparasion were made, fisrt one, PYR/PYL sextuple mutant versus Col-0 reference sample and second one, SnRK2 triple mutant versus Col-0 reference sample. In each comparasion four biological replicates were made. Replicas number 1 and 2 were labeled with Cy5 for the mutant sample and Cy3 for the Col-0 reference sample, while the other two replicas,#3 and #4, were reversed-labeled.

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Expression data from stratified anac089-2 mutant and wild-type seeds, ABA-treated.

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