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An ER-targeted calcium-binding peptide confers salt and drought tolerance mediated by CIPK6 in Arabidopsis

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Abstract

Different plant organelles have high internal stores of Ca2+ compared to the cytoplasm and could play independent roles in stress responses or signal transduction. We used a GFP fusion with the C-domain of calreticulin, which shows low-affinity, high capacity Ca2+ binding in the ER, as a calcium-binding peptide (CBP) to specifically increase stores in the ER and nucleus. Despite the presence of a signal sequence and KDEL retention sequence, our work and previous studies (Brandizzi et al. Plant Journal 34:269–281, 2003) demonstrated both ER and nuclear localization of GFP-CBP. Under normal conditions, GFP-CBP-expressing lines had ~25% more total Ca2+ and higher levels of chlorophyll and seed yield than wild type and GFP controls. CBP-expressing plants also had better survival under intermittent drought or high salt treatments and increased root growth. One member of the CIPK (calcineurin B-like interacting protein kinase) gene family, CIPK6, was up-regulated in CBP-expressing plants, even under non-stress conditions. A null mutation in cipk6 abolished the increased stress tolerance of CBP-transgenic plants, as well as the CBP-mediated induction of two stress-associated genes, DREB1A and RD29A, under non-stress conditions. Although this suggested that it was the induction of CIPK6, rather than localized changes in Ca2+, that resulted in increased survival under adverse conditions, CIPK6 induction still required Ca2+. This work demonstrates that ER (or nuclear) Ca2+ can directly participate in signal transduction to alter gene expression. The discovery of a method for increasing Ca2+ levels without deleterious effects on plant growth may have practical applications.

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Abbreviations

Ca2+ :

Calcium

CBL:

Calcineurin B-like

CIPK:

CBL interacting protein kinase

CBP:

Calcium-binding peptide

CRT:

Calreticulin

ER:

Endoplasmic reticulum

GFP:

Green fluorescent protein

SOS:

Salt overly sensitive

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Acknowledgments

We thank Dr. Rebecca Boston for the BiP and CRT antibodies, Dr. Wayne Robarge for analytical ICP readings, and Dr. Bill Hoffman for help with measuring stomatal conductance. We also thank Dr. Wendy Boss for her constructive comments and critical reading of the manuscript. GFP fluorescence was imaged in the NCSU Cell and Molecular Imaging Facility, and we thank Shantha Sumanasinghe for help. This research was supported by NASA grant # NAGW-4984, the NC Agricultural Research Service and the supported from the Korean Education Foundation.

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Authors and Affiliations

  1. Department of Cell and Molecular Biology, Grand Valley State University, Allendale, MI , 49401, USA

    Pei-Lan Tsou

  2. Department of Plant Biology, North Carolina State University, Box 7612, Raleigh, NC, 27695, USA

    Sang Yoon Lee, Nina Stromgren Allen, Heike Winter-Sederoff & Dominique Robertson

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  1. Pei-Lan Tsou
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  2. Sang Yoon Lee
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  3. Nina Stromgren Allen
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  4. Heike Winter-Sederoff
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  5. Dominique Robertson
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Correspondence to Dominique Robertson.

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P.-L. Tsou and S. Y. Lee contributed equally to this work.

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Tsou, PL., Lee, S.Y., Allen, N.S. et al. An ER-targeted calcium-binding peptide confers salt and drought tolerance mediated by CIPK6 in Arabidopsis . Planta 235, 539–552 (2012). https://doi.org/10.1007/s00425-011-1522-9

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