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Ectopic expression of CaRLK1 enhances hypoxia tolerance with increasing alanine production in Nicotiana spp.

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Abstract

In a previous report, the pepper receptor-like kinase 1 (CaRLK1) gene was shown to be responsible for negatively regulating plant cell death caused by pathogens via accumulation of superoxide anions. Here, we examined whether this gene also plays a role in regulating cell death under abiotic stress. The total concentrations of free amino acids in CaRLK1-overexpressed cells (RLKox) increased by twofold compared with those of the wild-type Nicotiana tabacum BY-2 cells. Additionally, alanine and pyruvate concentrations increased by approximately threefold. These accumulations were associated with both the expression levels of the isocitrate lyase (ICL) and malate synthase genes and their specific activities, which were preferentially up-regulated in the RLKox cells. The expression levels of ethylene biosynthetic genes (ACC synthase and ACC oxidase) were suppressed, but those of both the metallothionein and lesion simulating disease 1 genes increased in the RLKox cells during submergence-induced hypoxia. The specific activity of catalase, which is involved in protecting ICL from reactive oxygen species, was also induced threefold in the RLKox cells. The primary roots of the transgenic plants that were exposed to hypoxic conditions grew at similar rates to those in normal conditions. We propose that CaRLK1 maintains a persistent hypoxia-resistant phenotype.

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Abbreviations

ACCO:

1-Aminocyclopropene-1-carboxylic acid oxidase

ACCS:

1-Aminocyclopropene-1-carboxylic acid synthase

ADH:

Alcohol dehydrogenase

Ala:

Alanine

AlaAT:

Alanine aminotransferase

AMT:

Ammonium transporter

BY-2:

Nicotiana tabacum L. cv Bright Yellow 2 cell

CaRLK1:

Capsicum annuum receptor-like kinase 1

CC:

Callus culture conditions

DO:

Dissolved oxygen

EDS1:

Enhanced disease susceptibility 1

GABA:

γ-Amino butyric acid

GABAT:

γ-Amino butyric acid transaminase

GLB1:

Nitrogen regulatory protein PII

L-012:

8-Amino-5-chloro-7-phenylpyridol[3,4-d]pyridazine-1,4(2H,3H)dione

LSD1:

Lesion simulating disease 1

MT:

Metallothionein

PAD4:

Phytoalexin-deficient 4

PDC:

Pyruvate decarboxylase complex

RBOH:

Respiratory burst oxidase homologs

ROS:

Reactive oxygen species

SC:

Suspension culture conditions

SSADH:

Succinate semialdehyde dehydrogenase

SUB1:

Submergence 1

VGB:

Vigabatrin

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Acknowledgments

We thank Prof. Ro Dong Park for his comments on the manuscript. We greatly thank Prof. Francois Simon-Plas for supplying us with the anti-NtRBOHD antibody. This report is dedicated to Prof. Jan A. D. Zeevaart, who was my mentor (D. J. Lee) and died from pancreatic cancer in 2009. This research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) and was funded by the Ministry of Education, Science and Technology (NRF-2012R1A1A2006934) and the Second Stage BK21 Project from the Ministry of Education, Science and Technology of the Republic of Korea.

Conflict of interest

The author that is responsible for the distribution of materials that are integral to the findings that were presented in this article is Dong Ju Lee (leedongju@gmail.com). D. J. L. and Y-T. C. designed the experiments and wrote the manuscript. D. M. K., S. H. C., J. Y. L., and G-W. C. conducted the experiments. D. J. L. analysed the data. The authors declare no competing financial interests.

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Author notes

  1. Dong Ju Lee

    Present address: Institute of Environmentally-Friendly Agriculture, Chonnam National University, 300 Youngbong-dong, Buk-gu, Gwangju, 500-757, Republic of Korea

Authors and Affiliations

  1. School of Biological Sciences and Technology, Higher Education Center for Bioregulator Research, Chonnam National University, 300 Youngbong-dong, Buk-gu, Gwangju, 500-757, Republic of Korea

    Dong Ju Lee, Youn-Tae Chi, Ji Young Lee & Go-woo Choi

  2. Department of Medical System Engineering, Gwangju Institute of Science and Technology, 123 Cheomdan-gwagiro, Buk-gu, Gwangju, 500-712, Republic of Korea

    Dong Min Kim

  3. Department of Biological Sciences, Chonnam National University, 300 Youngbong-dong, Buk-gu, Gwangju, 500-757, Republic of Korea

    Seung-Hyun Choi

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Lee, D.J., Chi, YT., Kim, D.M. et al. Ectopic expression of CaRLK1 enhances hypoxia tolerance with increasing alanine production in Nicotiana spp.. Plant Mol Biol 86, 255–270 (2014). https://doi.org/10.1007/s11103-014-0227-4

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