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Proteomic analysis of reactive oxygen species (ROS)-related proteins in rice roots

  • Biotic and Abiotic Stress
  • Published:
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Abstract

To investigate the rice root proteome, we applied the PEG fractionation technique combined with two-dimensional gel electrophoresis which rendered more well-separated protein spots. Out of the 295 chosen proteins, 93 were identified by MALDI-TOF mass spectrometry. The proteins were classified as relating to metabolism (38.7%), reactive oxygen species (ROS)-related proteins (22.5%), protein processing/degradation (8.6%), stress/defense (7.5%), energy (6.5%) and signal transduction (5.4%). The high percentage of ROS-related proteins found in rice root brings us to assess the roles of ROS on rice root growth. Treatment with ROS quenching chemicals such as reduced glutathione (GSH), diphenyleneiodonium (DPI) and ascorbate inhibited root growth dose-dependently. Forty-nine proteins identified were either up- or down-regulated by GSH treatment, of which 14 were ROS-related proteins, such noticeably modulated ones as glutathione-S-transferase (GST), superoxide dismutases (SOD) and l-ascorbate peroxidases. The protein levels of four GSTs (NS4, 8, 56 and 57), three APXs (NS46, 49 and 50) and MnSOD (NS45) were strongly reduced by GSH treatment but slightly reduced by ascorbate and DPI. Ascorbate and DPI strongly inhibited expression levels of a catalase A (NP23) and an APX (NS65) but did not affect APXs (NS46, 49 and 50) protein levels. Northern analysis demonstrated that changes in transcript levels of five genes––GST (NS4), GST (NS43), Mn-SOD (NS45), APX (NS50) and APX (NS46/49) in response to ROS quenching chemicals were coherent with patterns shown in two-dimensional electrophoresis analyses. Taken together, we suggest that these proteins may take part in an important role in maintaining cellular redox homeostasis during rice root growth.

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Acknowledgments

This work was supported by Grant No. CG1122 from the Crop Functional Genomic Center; by a grant from KOSEF/MOST to the Environmental Biotechnology National Core Research Center (to S.G. Kim and S.T. Kim); and by scholarships from the Brain Korea 21 Program, Ministry of Education and Human Resources Development, Korea (to S.G. Kim, Y. Wang).

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Authors and Affiliations

  1. Division of Applied Life Science (BK21 program), Plant Molecular Biology and Biotechnology Research Center, Jinju, South Korea

    Sang Gon Kim, Sun Young Kang, Yiming Wang & Kyu Young Kang

  2. Environmental Biotechnology National Core Research Center, Gyeongsang National University, Jinju, 660-701, South Korea

    Sang Gon Kim, Sun Tae Kim & Kyu Young Kang

  3. College of Life Sciences and Biotechnology, Division of Biotechnology, Korea University, Anam-dong, Seoul, 136-713, South Korea

    Wook Kim

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  1. Sang Gon Kim
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Correspondence to Kyu Young Kang.

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Communicated by W.T. Kim.

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Kim, S.G., Kim, S.T., Kang, S.Y. et al. Proteomic analysis of reactive oxygen species (ROS)-related proteins in rice roots. Plant Cell Rep 27, 363–375 (2008). https://doi.org/10.1007/s00299-007-0441-5

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