Dataset Information

Responses of PYR/PYL/RCAR ABA Receptors to Contrasting stresses, Heat and Cold in Arabidopsis.

ABSTRACT: Plants growing in natural habitats have evolved a wide range of mechanisms to copy with environmental challenging, including biotic and abiotic stresses. Abiotic stresses-induced increases in Abscisic acid (ABA) levels in plants suffering from stresses, including drought, cold or heat stress. To explore the function of the core components in ABA signaling, we used the overexpression of RCARs transgenic plants to expose in heat or cold stress. In this study, overexpression of RCAR12 or RCAR13 (R12-OE or R13-OE) transgenic plants had higher germination and survival rate than the wild-type (WT) Arabidopsis, indicating that they are both positively responsive to the high temperature. And the heat shock genes HSP18.2 and HSP70 were significantly induced by RCAR12 or RCAR13. Further, the results inferred that the over-expression of RCAR12 or RCAR13 could tolerance the cold stress, through induction CBFs expressions, the cold-responsive genes when plants were challenged the cold tress. And when complementation of RCAR12 to the 1124 mutant (R12:1124), the results indicated that RCAR12 could recover the insensitivity of 1124 to heat and cold stresses. Hence, we propose that RCAR12 and RCAR13, the ABA receptors, may play the positive roles in regulating the extreme temperature, including cold and high temperature in Arabidopsis.

SUBMITTER: Zhang Q

PROVIDER: S-EPMC6866694 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature

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Responses of PYR/PYL/RCAR ABA Receptors to Contrasting stresses, Heat and Cold in Arabidopsis.

Zhang Qian Q Kong Xiangge X Yu Qin Q Ding Yongqiang Y Li Xiaoyi X Yang Yi Y

Plant signaling & behavior 20190925 12

Plants growing in natural habitats have evolved a wide range of mechanisms to copy with environmental challenging, including biotic and abiotic stresses. Abiotic stresses-induced increases in Abscisic acid (ABA) levels in plants suffering from stresses, including drought, cold or heat stress. To explore the function of the core components in ABA signaling, we used the overexpression of <i>RCARs</i> transgenic plants to expose in heat or cold stress. In this study, overexpression of <i>RCAR12</i> ...[more]

PMID: 31552801

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Project description:ABA signaling relies on the perception of the hormone by 14 different members of the PYR/PYL/RCAR family of ABA receptors. Upon ABA binding, receptors become active in inhibiting a family of type 2C protein phosphatases that usually function as negative regulators on protein kinases of the SnRK2 family. Previous physiological and genetic data supporting a negative role exerted by nitric oxide (NO) on ABA signaling have been reported. Our preliminary data suggested that negative regulation exerted by NO on ABA signaling may involve direct effects of NO on ABA receptors. Here we explore whether the effects of NO on ABA signaling are due to post-translational modifications (PTMs) on ABA receptors. Two main PTMs are functionally related to NO, the S-nitrosylation of C residues and the nitration of Y residues and they have been studied in this work. Because our data suggest that NO can also modulate the stability of the receptor proteins, we have also analyzed another PTM, the ubiquitination of K residues. A double approach based on in vitro nitration of recombinant PYR/PYL/RCAR proteins as well as in planta expression of HA-tagged versions of the receptors, followed by immunopurification strategies based on anti-3-nitroY-coated magnetic beads, and LC-MS/MS analyses of the immunopurified proteins under non-reducing conditions allowed us to identify peptides containing nitroY (mass shifts of +45), S-nitrosylated C (mass shifts of +29), and K-GG tags resulting from the trypsin digestion of ubiquitin tails (mass shift of + 114). Our data suggest that S-nitrosylation of cysteines from PYR/PYL receptors occur in planta but has not effect on receptor function as tested in vitro. By contrast, the nitration of several Y residues of PYR/PYL receptors completely inactivates them, promotes also their polyubiquitination and marks them for subsequent proteasomal degradation, thus representing a rapid mechanism for lowering ABA sensitivity when required in the plant.

Project description:BackgroundAbscisic acid (ABA) is an important phytohormone for plant growth, development and responding to stresses such as drought, salinity, and pathogen infection. Pyrabactin Resistance 1 (PYR1)/PYR1-Like (PYL)/Regulatory Component of ABA Receptor (RCAR) (hereafter referred to as PYLs) has been identified as the ABA receptors. The PYL family members have been well studied in many plants. However, the members of PYL family have not been systematically identified at genome level in cultivated tobacco (Nicotiana tabacum) and its two ancestors. In this study, the phylogenic relationships, chromosomal distribution, gene structures, conserved motifs/regions, and expression profiles of NtPYLs were analyzed.ResultsWe identified 29, 11, 16 PYLs in the genomes of allotetraploid N. tabacum, and its two diploid ancestors N. tomentosiformis and N. sylvestris, respectively. The phylogenetic analysis revealed that NtPYLs can be divided into three subfamilies, and each NtPYL has one counterpart in N. sylvestris or N. tomentosiformis. Based on microarray analysis of NtPYL transcripts, four NtPYLs (from subfamily II, III), and five NtPYLs (from subfamily I) are highlighted as potential candidates for further functional characterization in N. tabacum seed development, response to ABA, and germination, and resistance to abiotic stresses, respectively. Interestingly, the expression profiles of members in the same NtPYL subfamily showed somehow similar patterns in tissues at different developmental stages and in leaves of seedlings under drought stress, suggesting particular NtPYLs might have multiple functions in both plant development and drought stress response.ConclusionsNtPYLs are highlighted for important functions in seed development, germination and response to ABA, and particular in drought tolerance. This work will not only shed light on the PYL family in tobacco, but also provides some valuable information for functional characterization of ABA receptors in N. tabacum.

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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Responses of PYR/PYL/RCAR ABA Receptors to Contrasting stresses, Heat and Cold in Arabidopsis.

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Responses of PYR/PYL/RCAR ABA Receptors to Contrasting stresses, Heat and Cold in Arabidopsis.

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