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Gel-based proteomics approach for detecting low nitrogen-responsive proteins in cultivated rice species

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Abstract

Nitrogen fertilization is essential for increasing rice production to meet the food demands of increasing world’s population. We established an in vivo hydroponic rice seedling culture system to investigate physio-biochemical/molecular responses of various rice japonica and indica cultivars to low nitrogen (N). Three-week-old seedlings grown in Yoshida’s nutrient solution manifested stable and reproducible symptoms, such as reduced shoot growth and length under low N. Out of 12 genetically selected cultivars, 11 cultivars showed varied degrees of growth reduction response to applied N (4 and 40 ppm N for treatment and control, respectively), whereas one cultivar (no. 12) showed similar growth as the control though its leaf width was smaller than control. Leaves of a representative low N-responsive cultivar (BG90-2) were sampled for revealing protein profiles between low and normal (control) N application by two-dimensional gel electrophoresis (2-DGE). Forty-one proteins were identified with MALDI-TOF-MS and nESI-LC-MS/MS. Assignment of proteins into major (energy metabolism, photosynthesis and oxidative stress) and minor functional categories, revealed many novel low N-responsive proteins, including those having energy/photosynthesis- and defense/stress- and iron homeostasis-related functions. Results suggest the usefulness of proteomics in identifying novel N-responsive proteins and may provide potential markers for rice response to low N.

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Abbreviations

APX:

Ascorbate peroxidase

CHAPS:

3-[(3-cholamidopropyl) dimethylaminol]-1-propanesulfonate

DTT:

dithiothreitol

LSU:

Large subunit

SOD:

Superoxide dismutase

SSU:

Small subunit

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Authors and Affiliations

  1. Korea Bio Polytechnic College, 315-1, Chaewoon-ri, Ganggyeong-eup, Nonsan-si, Chungnam, 320-905, Korea

    Du Hyun Kim

  2. Health Technology Research Center (HTRC), National Institute of Advanced Industrial Science and Technology (AIST) West, Tsukuba, Ibaraki, 305-8569, Japan

    Junko Shibato, Hitoshi Iwahashi, Yoshinori Masuo & Randeep Rakwal

  3. National Institute of Crop Science, Rural Development Administration (RDA), Suwon, 441-100, Korea

    Dea-Wook Kim, Myung Kyu Oh & Myung Ki Kim

  4. Department of Environmental Horticulture, University of Seoul, Seoul, 130-743, Korea

    Ie Sung Shim

  5. Research Laboratory for Biotechnology and Biochemistry (RLABB), Kathmandu, Nepal

    Randeep Rakwal

Authors
  1. Du Hyun Kim
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  2. Junko Shibato
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  3. Dea-Wook Kim
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  4. Myung Kyu Oh
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  5. Myung Ki Kim
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  6. Ie Sung Shim
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  7. Hitoshi Iwahashi
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  8. Yoshinori Masuo
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  9. Randeep Rakwal
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Correspondence to Randeep Rakwal.

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Kim, D.H., Shibato, J., Kim, DW. et al. Gel-based proteomics approach for detecting low nitrogen-responsive proteins in cultivated rice species. Physiol Mol Biol Plants 15, 31–41 (2009). https://doi.org/10.1007/s12298-009-0003-0

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  • DOI: https://doi.org/10.1007/s12298-009-0003-0

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