Identification of Cu-binding proteins in embryos of germinating rice in response to Cu toxicity

  • Hongxiao Zhang
  • Yufeng Song
  • Fayuan Wang
  • Youjun Li
  • Hui Wang
  • Liming YangEmail author
Original Article


Seed germination, an early and important process for the growth and development of plants, is hypersensitive to environmental changes. Copper (Cu) is a necessary micronutrient for plants; however, an excessive dose of Cu had an extremely negative effect at the cellular level as a result of inevitable binding to proteins. In contrast, some structural motifs of proteins can bind free Cu ions and relieve Cu toxicity. This study aimed to understand the expression characteristics of Cu-binding proteins induced by excess Cu during rice seed germination. We investigated Cu-binding proteins in germinating rice embryos treated with 200 µM Cu using a Sephadex G-50 column or immobilized Cu affinity chromatography combined with two-dimensional gel electrophoresis. Proteomics analysis indicated that 12 protein spots exhibited a > 2.0-fold increase in intensity in response to Cu toxicity as compared with controls. Among nine proteins in ten spots identified as Cu-binding proteins, three proteins (from four spots) were involved in antioxidative defense: copper, zinc superoxide dismutase, glutathione S-transferase and protein disulfide isomerase. These results show that reactive oxygen species may be involved in the expression regulation of Cu-binding proteins in germinating rice in response to Cu stress.


Cu stress Rice seed germination Cu-binding protein Immobilized Cu affinity chromatography Proteomics 



Two-dimensional gel electrophoresis


Cu-binding protein


Immobilized Cu affinity chromatography


Copper, zinc superoxide dismutase




Glutathione S-transferase


Iminodiacetic acid


Immobilized metal affinity chromatography


Jacalin-related lectin


Protein disulfide isomerase


Phenylmethylsulfonyl fluoride



This research project was partly supported by the Key Projects of the Department of Education of Henan Province (16A180004), the Key Projects of Science and Technology of Henan Province (21010335), Innovation Team Foundation of Henan University of Science and Technology (2015TTD002) and Jiangsu Government Scholarship for Overseas Studies.


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Copyright information

© Franciszek Górski Institute of Plant Physiology, Polish Academy of Sciences, Kraków 2018

Authors and Affiliations

  • Hongxiao Zhang
    • 1
  • Yufeng Song
    • 2
    • 3
  • Fayuan Wang
    • 1
    • 4
  • Youjun Li
    • 1
  • Hui Wang
    • 5
  • Liming Yang
    • 2
    Email author
  1. 1.College of AgricultureHenan University of Science and TechnologyLuoyangChina
  2. 2.College of Biology and the EnvironmentNanjing Forestry UniversityNanjingChina
  3. 3.Liaoning Jinda Shengyuan GroupCoastal New Industrial ZoneYingkouChina
  4. 4.College of Environment and Safety EngineeringQingdao University of Science and TechnologyQingdaoChina
  5. 5.Department of Plant PathologyUniversity of GeorgiaTiftonUSA

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