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Identification and characterization of a novel chloroplast/mitochondria co-localized glutathione reductase 3 involved in salt stress response in rice

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Abstract

Glutathione reductases (GRs) are important components of the antioxidant machinery that plants use to respond against abiotic stresses. In rice, one cytosolic and two chloroplastic GR isoforms have been identified. In this work, we describe the cloning and characterization of the full-length cDNA encoding OsGR3, a chloroplast-localized GR that up to now was considered as a non-functional enzyme because of assumed lack of N-terminal conserved domains. The expression of OsGR3 in E. coli validated that it can be translated as a protein with GR activity. OsGR3 shows 76 and 53 % identity with OsGR1 (chloroplastic) and OsGR2 (cytosolic), respectively. Phylogenetic analysis revealed 2 chloroplastic GRs in Poaceae species, including rice, sorghum and brachypodium, but only one chloroplastic GR in dicots. A plastid transit peptide is located at the N terminus of OsGR3, and genetic transformation of rice with a GR3-GFP fusion construct further confirmed its localization in chloroplasts. Furthermore, OsGR1 and OsGR3 are also targeted to mitochondria, which suggest a combined antioxidant mechanism in both chloroplasts and mitochondria. However, both isoforms showed a distinct response to salinity: the expression of OsGR3 but not OsGR1 was induced by salt stress. In addition, the transcript level of OsGR3 was greatly increased with salicylic acid treatment but was not significantly affected by methyl jasmonate, dehydration or heat shock stress. Our results provide new clues about the possible roles of functional OsGR3 in salt stress and biotic stress tolerance.

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Acknowledgments

We thank Laura Smales for English editing. We are grateful to the Joint Center for Instruments and Researches of the College of Bioresources and Agriculture at National Taiwan University for confocal microscopy and technical support. This work was supported by research grants from the National Science Council of the Republic of China (NSC 98-2324-B-002-002 and NSC 98-2313-B-002-015-MY3) to C.-Y. Hong.

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Authors and Affiliations

  1. Department of Agricultural Chemistry, National Taiwan University, Taipei, 10617, Taiwan

    Tsung-Meng Wu, Wan-Rong Lin, Yun-Ting Kao & Chwan-Yang Hong

  2. Department of Agronomy, National Chung-Hsing University, Taichung, 40227, Taiwan

    Yi-Ting Hsu

  3. Department of Life Sciences, National Central University, Jhong-li, 32001, Taoyuan, Taiwan

    Ching-Hui Yeh

  4. Department of Agronomy, National Taiwan University, Taipei, 10617, Taiwan

    Ching Huei Kao

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  1. Tsung-Meng Wu
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Correspondence to Chwan-Yang Hong.

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11103_2013_95_MOESM1_ESM.ppt

Fig S1. Alignment of the 5’ RACE product with truncated cDNA and the final assembled OsGR3 gene. DNA sequence alignment for OsGR3 5’RACE product, truncated OsGR3 cDNA (Tc-OsGR3; accession no. AK108799) from databases and final assembled gene (this study). Identical residues with the final assembled gene (GR3 cDNA) are on a gray background. DNA sequence overlap between the 5’ RACE product, Tc-OsGR3 and the final assembled gene are on a black background. The alignment was generated by use of Clustal W 2.0 (Larkin et al., 2007) (PPT 824 kb)

11103_2013_95_MOESM2_ESM.ppt

Fig S2. Alignment of the OsGR3 complete coding sequence with rice genome sequences. Identical residues with exon sequences are in white on a black background. Identical residues with intron sequences are in black on a gray background. The alignment was generated by use of Clustal W 2.0 (Larkin et al., 2007) (PPT 1799 kb) (PPT 1099 kb)

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Wu, TM., Lin, WR., Kao, YT. et al. Identification and characterization of a novel chloroplast/mitochondria co-localized glutathione reductase 3 involved in salt stress response in rice. Plant Mol Biol 83, 379–390 (2013). https://doi.org/10.1007/s11103-013-0095-3

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