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Suppression of Arabidopsis AtPUB30 resulted in increased tolerance to salt stress during germination

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Abstract

Key message

The Arabidopsis U-box E3 Ub ligase AtPUB30 participates in the salt stress tolerance as a negative factor in an ABA-independent manner during germination.

Abstract

Based on the in silico expression data, the U-box protein 30 (AtPUB30) from Arabidopsis thaliana was identified as a gene that responds to salt stress. The deduced AtPUB30 protein consists of 448 amino acids with a single U-box motif and five ARM-repeat domains. An in vitro self-ubiquitination assay demonstrated that bacterially expressed AtPUB30 exhibited E3 ubiquitin (Ub) ligase activity and that the U-box domain was essential for the activity. Real-time qRT-PCR and promoter-GUS analyses showed that AtPUB30 was induced by high salinity, but not by drought, cold, or abscisic acid (ABA), in roots but not in shoots. These results suggest that AtPUB30 is an Arabidopsis U-box E3 Ub ligase, the expression of which is selectively enhanced by salt stress in roots. T-DNA-inserted loss-of-function atpub30 mutant plants (atpub30-1 and atpub30-2) were more tolerant to salt stress in the germination stage, as identified by radicle emergence, cotyledon opening, and more vigorous early root growth relative to wild-type plants. Thus, it is likely that AtPUB30 plays a negative role in high salinity tolerance in the germination process. Wild type and mutant plants displayed very similar germination rates when treated with ABA, suggesting that the action of AtPUB30 in the germination stage is ABA independent. The post-germination growth of NaCl-stressed wild type and mutant plants were indistinguishable. Overall, our data suggest that the Arabidopsis U-box E3 Ub ligase AtPUB30 participates in the salt stress tolerance as a negative factor in the germination stage in root tissues.

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Abbreviations

ABA:

Abscisic acid

ARM:

Repeats armadillo repeats

CaMV:

Cauliflower mosaic virus

GFP:

Green fluorescent protein

HECT:

Homology to E6-AP carboxyl terminus

MBP:

Maltose-binding protein

RING:

Really interesting new gene

Ub:

Ubiquitin

UPS:

Ubiquitin-26S proteasome system

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Acknowledgments

This work was supported by grants from the National Research Foundation (Project No. 2010-0000782 and No. 2014-003891) and from the National Center for GM Crops (Project No. PJ008152) of the Next Generation BioGreen 21 Program funded by the Rural Development Administration, Republic of Korea, to W.T.K. and from the Korea Institute of Science and Technology Information to B.G.K.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Systems Biology, College of Life Science and Biotechnology, Yonsei University, Seoul, 120-749, Korea

    Jae Hwan Hwang, Dong Hye Seo & Woo Taek Kim

  2. ReSEAT Program, Korea Institute of Science and Technology Information, Seoul, 130-741, Korea

    Bin Goo Kang

  3. Department of Cellular Biology and Molecular Genetics, University of Maryland, College Park, MD, 20742, USA

    June M. Kwak

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  1. Jae Hwan Hwang
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  2. Dong Hye Seo
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Correspondence to Woo Taek Kim.

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Communicated by Jeong Sheop Shin.

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Hwang, J.H., Seo, D.H., Kang, B.G. et al. Suppression of Arabidopsis AtPUB30 resulted in increased tolerance to salt stress during germination. Plant Cell Rep 34, 277–289 (2015). https://doi.org/10.1007/s00299-014-1706-4

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