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Overexpression of BrCIPK1 Gene Enhances Abiotic Stress Tolerance by Increasing Proline Biosynthesis in Rice

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Abstract

The calcineurin B-like protein (CBL)-CBL-interacting protein kinase (CIPK) pathway is emerging as a major signaling pathway in plants. To understand the function of CIPK, the gene named BrCIPK1 from Brassica rapa were introduced into rice. Characterization of BrCIPK1 gene showed a 1982 bp, containing 1509 bp coding region and 502 amino acids. Green fluorescent protein (GFP)-tagged BrCIPK1 was observed exclusively in the cytoplasmic and peripheral regions in the plant cell. Gene expression showed that its messenger RNA (mRNA) transcription in B. rapa was differentially accumulated in the presence of cold, salinity, and drought, indicating its biological roles in multiple stress response pathways in plants. Furthermore, Ubi-1::BrCIPK1 rice lines showed significantly higher biomass, water content, and proline and free sugar content relative to those in the wild-type Gopum. The BrCIPK1 interacted with rice calcineurin B-like protein 1 and 5 (OsCBL1, OsCBL5), suggesting that it is activated by Ca2+-bound CBLs in the cytosol by calcium spiking and regulates its downstream target proteins in these regions to increase abiotic stress tolerance. The results imply that BrCIPK1 gene may be involved in stress adaptations through the activation of pyrroline-5-carboxylate synthase in the proline biosynthetic pathway. In this paper, a hypothetical mechanism of elevated tolerance to cold, drought, and salinity is presented.

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Acknowledgments

This work was supported by Chungbuk National University in 2013, Golden Seed Project, Ministry of Agriculture, Food and Rural Affairs (MAFRA), and the National Research Foundation (NRF) programs (2014R1A2A1A11052547), the Korean Ministry of Science, ICT and Future Planning, Republic of Korea.

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Author notes

  1. Sailila E. Abdula

    Present address: Philippine Rice Research Institute-Midsayap, North Cotabato, 9410, Philippines

Authors and Affiliations

  1. Department of Crop Science, Chungbuk National University, Cheongju, 362-763, Korea

    Sailila E. Abdula, Hye-Jung Lee & Yong-Gu Cho

  2. Department of Biology, Chungbuk National University, Cheongju, 362-763, Korea

    Hojin Ryu

  3. Department of Horticulture, Hankyong National University, Ansung, 456-749, Korea

    Kwon Kyoo Kang

  4. Department of Horticulture, Sunchon National University, Sunchon, 540-742, Korea

    Illsup Nou

  5. Department of Plant Breeding and Genetics, Cornell University, Ithaca, NY, 14853, USA

    Mark E. Sorrells

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  1. Sailila E. Abdula
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Correspondence to Yong-Gu Cho.

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Sailila E. Abdula and Hye-Jung Lee contributed equally to this work.

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Fig. S1

Schematic diagram of the binary Ti plasmid pSB11 containing the CBL interacting protein kinase 1 full length cDNA from Chinese cabbage. P35S, CaMV35S promoter; pUbi-1, maize Ubiquitin-1 promoter; Tg7 and Tnos, polyadenylation signals from gene 7 and nopaline synthase (nos) gene in the T-DNA, respectively; HPT, hygromycin phosphotransferase gene; LB, left border; RB, right border (gif 8 kb)

High Resolution Image (tiff 1678 kb)

Fig. S2

Genomic and expression of Ubi-1::BrCIPK1 rice lines and wild type Gopum. (a) PCR Confirmation of BrCIPK1 insert in rice using BrCIPK1 and HPT primers in 1 % agarose gel. WT, Gopum; P, control plasmid DNA. (b) Relative mRNA expression of BrCIPK1 in transgenic rice and wild type (gif 44 kb)

High Resolution Image (tiff 5393 kb)

Fig. S3

Respond of Ubi-1::BrCIPK1 rice lines under stress condition. Phenotype of BrCIPK1 relative to susceptible check and wild type. (a) Cold stress at 10 °C. T (tolerant variety), Seorak; S (susceptible variety), Gaya. (b) Salinity stress with 130 mM NaCl. T (tolerant variety), Hwayeong; MT (moderate tolerant variety), Junam; S (susceptible variety), Dongjin. (c) Drought stress with 20 % PEG6000. T (tolerant variety), Sangnambat; S (susceptible variety), Gaya; WT (wild type), Gopum (gif 305 kb)

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Abdula, S.E., Lee, HJ., Ryu, H. et al. Overexpression of BrCIPK1 Gene Enhances Abiotic Stress Tolerance by Increasing Proline Biosynthesis in Rice. Plant Mol Biol Rep 34, 501–511 (2016). https://doi.org/10.1007/s11105-015-0939-x

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