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Plant Molecular Biology

, Volume 89, Issue 1–2, pp 143–156 | Cite as

Functional roles of the pepper RING finger protein gene, CaRING1, in abscisic acid signaling and dehydration tolerance

  • Chae Woo Lim
  • Byung Kook Hwang
  • Sung Chul Lee
Article

Abstract

Plants are constantly exposed to a variety of biotic and abiotic stresses, which include pathogens and conditions of high salinity, low temperature, and drought. Abscisic acid (ABA) is a major plant hormone involved in signal transduction pathways that mediate the defense response of plants to abiotic stress. Previously, we isolated Ring finger protein gene (CaRING1) from pepper (Capsicum annuum), which is associated with resistance to bacterial pathogens, accompanied by hypersensitive cell death. Here, we report a new function of the CaRING1 gene product in the ABA-mediated defense responses of plants to dehydration stress. The expression of the CaRING1 gene was induced in pepper leaves treated with ABA or exposed to dehydration or NaCl. Virus-induced gene silencing of CaRING1 in pepper plants exhibited low degree of ABA-induced stomatal closure and high levels of transpirational water loss in dehydrated leaves. These led to be more vulnerable to dehydration stress in CaRING1-silenced pepper than in the control pepper, accompanied by reduction of ABA-regulated gene expression and low accumulation of ABA and H2O2. In contrast, CaRING1-overexpressing transgenic plants showed enhanced sensitivity to ABA during the seedling growth and establishment. These plants were also more tolerant to dehydration stress than the wild-type plants because of high ABA accumulation, enhanced stomatal closure and increased expression of stress-responsive genes. Together, these results suggest that the CaRING1 acts as positive factor for dehydration tolerance in Arabidopsis by modulating ABA biosynthesis and ABA-mediated stomatal closing and gene expression.

Keywords

Abscisic acid CaRING1 Dehydration Pepper Transgenic plant 

Abbreviations

ABA

Abscisic acid

Pst

Pseudomonas syringae pv. tomato

PR

Pathogenesis related

qRT-PCR

Quantitative reverse transcription-polymerase chain reaction

ROS

Reactive oxygen species

SA

Salicylic acid

SOS

Stomatal opening solution

VIGS

Virus-induced gene silencing

Xcv

Xanthomonas campestris pv. vesicatoria

Notes

Acknowledgments

This work was supported by a Grant from “The Next-Generation BioGreen 21 Program for Agriculture & Technology Development (Project No. PJ011010501)” Rural Development Administration, Republic of Korea.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Human and animal rights

Human participants and/or animals have not involved in this research.

Supplementary material

11103_2015_359_MOESM1_ESM.pptx (54 kb)
Supplementary material 1 (PPTX 54 kb)
11103_2015_359_MOESM2_ESM.pptx (625 kb)
Supplementary Fig. 1 Expression patterns of CaRING1-homologous genes from Arabidopsis in response to ABA and drought stress. (A and B) Relative expression levels of Arabidopsis RING-H2 genes in guard cells and leaves of 5-week-old plants 3 h after treatment with 50 μM ABA or ethanol (control). (C and D) Relative expression levels of Arabidopsis RING-H2 genes in 7-day-old seedling treated with 10 μM ABA. (E and F) Relative expression levels of Arabidopsis RING-H2 genes in 18-day-old seedling treated with drought stress. For drought stress treatment, plants grown on rafts in Magenta boxes were exposed to the air stream for 15 min with loss of approximately 10 % fresh weight. Data were obtained from Arabidopsis eFP Browser in the Bio-Analytic Resource for Plant Biology (http://bar.utoronto.ca/efp/cgi-bin/efpWeb.cgi). (PPTX 624 kb)
11103_2015_359_MOESM3_ESM.pptx (82 kb)
Supplementary Fig. 2 Expression patterns of CaRING1-homologous genes from Arabidopsis in response to cold, osmotic, and salt stress. Relative expression levels of Arabidopsis RING-H2 genes in shoot (right) and root (left) of 18-day-old seedling treated with cold (A; continuous 4 °C on crushed ice in cold chamber), osmotic (B; 300 mM mannitol), and salt (C; 150 mM NaCl). Data were obtained from Arabidopsis eFP Browser in the Bio-Analytic Resource for Plant Biology (http://bar.utoronto.ca/efp/cgi-bin/efpWeb.cgi). (PPTX 81 kb)
11103_2015_359_MOESM4_ESM.pptx (54 kb)
Supplemental Fig. S3 Constitutive expression of CaRING1 in the transgenic lines. RT-PCR analysis of CaRING1 gene expression in wild-type and transgenic lines. Actin 8 was used as an internal control. (PPTX 53 kb)
11103_2015_359_MOESM5_ESM.pptx (126 kb)
Supplemental Fig. S4 Effect of norflurazon (NF) and ABA during seed germination and seedling establishment of 35S:CaRING1 mutants. The seeds of 35S:CaRING1 mutants and wild-type plants were germinated on 0.5X MS containing 50 μM NF alone or 50 μM NF and 0.75 μM ABA. After 10 days, the representative images were taken. (PPTX 125 kb)
11103_2015_359_MOESM6_ESM.pptx (1016 kb)
Supplemental Fig. S5 ABA and dehydration sensitivity of rfh2 mutants. (A) Scheme of the Arabidopsis RFH2 gene structure. White and black boxes indicate exons and introns, respectively. The positions of the T-DNA insertions in rhf2 mutants are indicated by triangles. (B) Seedling growth of wild-type and rhf2 mutants exposed to ABA. The seedlings were grown vertically in 0.5X MS containing 0.75 μM ABA for 7 days. The representative images were taken. (C) Dehydration sensitivity of the rfh2 mutants. Three-week-old wild-type (WT) and rfh2 mutants were treated with dehydration stress by withholding water for 14 days, followed by rehydration for 3 days. The representative images were taken and percentages of plants that survived were measured. (PPTX 1015 kb)

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Copyright information

© Springer Science+Business Media Dordrecht 2015

Authors and Affiliations

  • Chae Woo Lim
    • 1
  • Byung Kook Hwang
    • 2
  • Sung Chul Lee
    • 1
  1. 1.Department of Life Science (BK21 Program)Chung-Ang UniversitySeoulKorea
  2. 2.School of Life Sciences and BiotechnologyKorea UniversitySeoulKorea

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