Advertisement

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 Yang
Original Article
  • 31 Downloads

Abstract

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.

Keywords

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

Abbreviations

2-DE

Two-dimensional gel electrophoresis

CuBP

Cu-binding protein

Cu-IMAC

Immobilized Cu affinity chromatography

Cu/Zn SOD

Copper, zinc superoxide dismutase

DTT

Dithiothreitol

GST

Glutathione S-transferase

IDA

Iminodiacetic acid

IMAC

Immobilized metal affinity chromatography

JRL

Jacalin-related lectin

PDI

Protein disulfide isomerase

PMSF

Phenylmethylsulfonyl fluoride

Notes

Acknowledgements

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.

References

  1. Ahsan N, Lee DG, Lee SH, Kang KY, Lee JJ, Kim PJ, Yoon HS, Kim JS, Lee BH (2007a) Excess copper induced physiological and proteomic changes in germinating rice seeds. Chemosphere 67:1182–1193CrossRefPubMedGoogle Scholar
  2. Ahsan N, Lee SH, Lee DG, Lee H, Lee SW, Bahk JD, Lee BH (2007b) Physiological and protein profiles alternation of germinating rice seedlings exposed to acute cadmium toxicity. C R Biol 330:735–746CrossRefPubMedGoogle Scholar
  3. Al Atalah B, Smagghe G, Van Damme EJM (2014) Orysata, a jacalin-related lectin from rice, could protect plants against biting-chewing and piercing-sucking insects. Plant Sci 221–222:21–28CrossRefPubMedGoogle Scholar
  4. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254CrossRefPubMedGoogle Scholar
  5. Chen Z, Pan Y, Wang S, Ding Y, Yang W, Zhu C (2012) Overexpression of a protein disulfide isomerase-like protein from Methanothermobacter thermoautotrophicum enhances mercury tolerance in transgenic rice. Plant Sci 197:10–20CrossRefPubMedGoogle Scholar
  6. Ciriolo MR, Civitareale P, Carri MT, De Martino A, Galiazzo F, Rotilio G (1994) Purification and characterization of Ag, Zn-superoxide dismutase from Saccharomyces cerevisiae exposed to silver. J Biol Chem 269:25783–25787PubMedGoogle Scholar
  7. Claes B, Dekeyser R, Villarroel R, Van den Bulcke M, Bauw G, Van Montagu M, Caplan A (1990) Characterization of a rice gene showing organ-specific expression in response to salt stress and drought. Plant Cell 2:19–27CrossRefPubMedPubMedCentralGoogle Scholar
  8. Haensch R, Mendel RR (2009) Physiological functions of mineral micronutrients (Cu, Zn, Mn, Fe, Ni, Mo, B, Cl). Curr Opin Plant Biol 12:259–266CrossRefGoogle Scholar
  9. Hatahet F, Ruddock LW (2009) Protein disulfide isomerase: a critical evaluation of its function in disulfide bond formation. Antioxid Redox Sign 11:2807–2850CrossRefGoogle Scholar
  10. He J, Ren Y, Pan X, Yan Y, Zhu C, Jiang D (2010) Salicylic acid alleviates the toxicity effect of cadmium on germination, seedling growth, and amylase activity of rice. J Plant Nutr Soil Sci 173:300–305CrossRefGoogle Scholar
  11. Holdsworth MJ, Bentsink L, Soppe WJJ (2008) Molecular networks regulating Arabidopsis seed maturation, after-ripening, dormancy and germination. New Phytol 179:33–54CrossRefPubMedGoogle Scholar
  12. Khatoon A, Rehman S, Hiraga S, Makino T, Komatsu S (2012) Organ-specific proteomics analysis for identification of response mechanism in soybean seedlings under flooding stress. J Proteomics 75:5706–5723CrossRefPubMedGoogle Scholar
  13. Kung CCS, Huang WN, Huang YC, Yeh KC (2006) Proteomic survey of copper-binding proteins in Arabidopsis roots by immobilized metal affinity chromatography and mass spectrometry. Proteomics 6:2746–2758CrossRefPubMedGoogle Scholar
  14. Lannoo N, Van Damme EJM (2010) Nucleocytoplasmic plant lectins. Biochim Biophys Acta (BBA)-Gen Subj 1800:190–201CrossRefGoogle Scholar
  15. Laurindo FRM, Pescatore LA, Fernandes DdC (2012) Protein disulfide isomerase in redox cell signaling and homeostasis. Free Radical Biol Med 52:1954–1969CrossRefGoogle Scholar
  16. Li F, Shi J, Shen C, Chen G, Hu S, Chen Y (2009) Proteomic characterization of copper stress response in Elsholtzia splendens roots and leaves. Plant Mol Biol 71:251–263CrossRefPubMedGoogle Scholar
  17. Li Y, Yang M, Li Y, Liu W, Wen J, Li Y (2011) Differential gene and protein expression in soybean at early stages of incompatible interaction with Phytophthora sojae. Agr Sci China 10:902–910CrossRefGoogle Scholar
  18. Lombardi L, Sebastiani L (2005) Copper toxicity in Prunus cerasifera: growth and antioxidant enzymes responses of in vitro grown plants. Plant Sci 168:797–802CrossRefGoogle Scholar
  19. Lou LQ, Shen ZG, Li XD (2004) The copper tolerance mechanisms of Elsholtzia haichowensis, a plant from copper-enriched soils. Environ Exp Bot 51:111–120CrossRefGoogle Scholar
  20. Nonogaki H, Bassel GW, Bewley JD (2010) Germination-still a mystery. Plant Sci 179:574–581CrossRefGoogle Scholar
  21. Porath J, Carlsson J, Olsson I, Belfrage G (1975) Metal chelate affinity chromatography, a new approach to protein fractionation. Nature 258:598–599CrossRefPubMedGoogle Scholar
  22. Qian M, Li XD, Shen ZG (2005) Adaptive copper tolerance in Elsholtzia haichowensis involves production of Cu-induced thiol peptides. Plant Growth Regul 47:65–73CrossRefGoogle Scholar
  23. Rajjou L, Duval M, Gallardo K, Catusse J, Bally J, Job C, Job D (2012) Seed germination and vigor. Annu Rev Plant Biol 63:507–533CrossRefPubMedGoogle Scholar
  24. Rhee SG, Woo HA, Kil IS, Bae SH (2012) Peroxiredoxin functions as a peroxidase and a regulator and sensor of local peroxides. J Biol Chem 287:4403–4410CrossRefPubMedGoogle Scholar
  25. Schuler MA, Werck-Reichhart D (2003) Functional genomics of P450s. Annu Rev Plant Biol 54:629–667CrossRefPubMedGoogle Scholar
  26. Schützendübel A, Polle A (2002) Plant responses to abiotic stresses: heavy metal-induced oxidative stress and protection by mycorrhization. J Exp Bot 53:1351–1365PubMedGoogle Scholar
  27. Smith SD, She Y-M, Roberts EA, Sarkar B (2004) Using immobilized metal affinity chromatography, two-dimensional electrophoresis and mass spectrometry to identify hepatocellular proteins with copper-binding ability. J Proteome Res 3:834–840CrossRefPubMedGoogle Scholar
  28. Song Y, Cui J, Zhang H, Wang G, Zhao FJ, Shen Z (2013) Proteomic analysis of copper stress responses in the roots of two rice (Oryza sativa L.) varieties differing in Cu tolerance. Plant Soil 366:647–658CrossRefGoogle Scholar
  29. Song M, Xu W, Xiang Y, Jia H, Zhang L, Ma Z (2014a) Association of jacalin-related lectins with wheat responses to stresses revealed by transcriptional profiling. Plant Mol Biol 84:95–110CrossRefGoogle Scholar
  30. Song Y, Zhang H, Chen C, Wang G, Zhuang K, Cui J, Shen Z (2014b) Proteomic analysis of copper-binding proteins in excess copper-stressed rice roots by immobilized metal affinity chromatography and two-dimensional electrophoresis. Biometals 27:265–276CrossRefPubMedGoogle Scholar
  31. Sun X, Chiu JF, He QY (2005) Application of immobilized metal affinity chromatography in proteomics. Expert Rev Proteomics 2:649–657CrossRefPubMedGoogle Scholar
  32. Sun X, Xiao C, Ge R, Yin X, Li H, Li N, Yang X, Zhu Y, He X, He QY (2011) Putative copper- and zinc-binding motifs in Streptococcus pneumoniae identified by immobilized metal affinity chromatography and mass spectrometry. Proteomics 11:3288–3298CrossRefPubMedGoogle Scholar
  33. Sunkar R, Kapoor A, Zhu JK (2006) Posttranscriptional induction of two Cu/Zn superoxide dismutase genes in Arabidopsis is mediated by downregulation of miR398 and important for oxidative stress tolerance. Plant cell 18:2051–2065CrossRefPubMedPubMedCentralGoogle Scholar
  34. Tan YF, O’Toole N, Taylor NL, Millar AH (2010) Divalent metal ions in plant mitochondria and their role in interactions with proteins and oxidative stress-induced damage to respiratory function. Plant Physiol 152:747–761CrossRefPubMedPubMedCentralGoogle Scholar
  35. Wang X, Kong H, Liu Z, Wang G (2012a) Expression analysis of genes encoding deoxycytidine/cytidine deaminases in rice. Chinese J Rice Sci 26:261–266Google Scholar
  36. Wang YD, Wang X, Wong Y (2012b) Proteomics analysis reveals multiple regulatory mechanisms in response to selenium in rice. J Proteomics 75:1849–1866CrossRefPubMedGoogle Scholar
  37. Wang C, Zhang X, Fan Y, Gao Y, Zhu Q, Zheng C, Qin T, Li Y, Che J, Zhang M (2015) XA23 is an executor R protein and confers broad-spectrum disease resistance in rice. Mol Plant 8:290–302CrossRefPubMedGoogle Scholar
  38. Yan JX, Wait R, Berkelman T, Harry RA, Westbrook JA, Wheeler CH, Dunn MJ (2000) A modified silver staining protocol for visualization of proteins compatible with matrix-assisted laser desorption/ionization and electrospray ionization-mass spectrometry. Electrophoresis 21:3666–3672CrossRefPubMedGoogle Scholar
  39. Yang L, Tian D, Todd CD, Luo Y, Hu X (2013) Comparative proteome analyses reveal that nitric oxide is an important signal molecule in the response of rice to aluminum toxicity. J Proteome Res 12:1316–1330CrossRefPubMedGoogle Scholar
  40. Yang JD, Worley E, Ma Q, Li J, Torres-Jerez I, Li G, Zhao PX, Xu Y, Tang Y, Udvardi M (2016) Nitrogen remobilization and conservation, and underlying senescence-associated gene expression in the perennial switch grass Panicum virgatum. New Phytol 211:75–89CrossRefPubMedGoogle Scholar
  41. Yruela I (2009) Copper in plants: acquisition, transport and interactions. Funct Plant Biol 36:409–430CrossRefGoogle Scholar
  42. Zhang H, Lian C, Shen Z (2009) Proteomic identification of small, copper-responsive proteins in germinating embryos of Oryza sativa. Ann Bot 103:923–930CrossRefPubMedPubMedCentralGoogle Scholar
  43. Zhang H, Zhang F, Xia Y, Wang G, Shen Z (2010) Excess copper induces production of hydrogen peroxide in the leaf of Elsholtzia haichowensis through apoplastic and symplastic CuZn-superoxide dismutase. J Hazar Mater 178:834–843CrossRefGoogle Scholar
  44. Zhang HX, Xia Y, Chen C, Zhuang K, Song Y, Shen ZG (2016) Analysis of copper-binding proteins in rice radicles exposed to excess copper and hydrogen peroxide stress. Front Plant Sci 7:1216PubMedPubMedCentralGoogle Scholar
  45. Zhang HX, Lv SF, Xu HW, Hou DY, Li YJ, Wang FY (2017) H2O2 is involved in the metallothionein-mediated rice tolerance to copper and cadmium toxicity. Int J Mol Sci 18:2083CrossRefPubMedCentralGoogle Scholar
  46. Zhao F, McGrath S, Crosland A (1994) Comparison of three wet digestion methods for the determination of plant sulphur by inductively coupled plasma atomic emission spectroscopy (ICP-AES). Commun Soil Sci Plant 25:407–418CrossRefGoogle Scholar
  47. Zhao L, Sun YL, Cui SX, Chen M, Yang HM, Liu HM, Chai TY, Huang F (2011) Cd-induced changes in leaf proteome of the hyperaccumulator plant Phytolacca americana. Chemosphere 85:56–66CrossRefPubMedGoogle Scholar

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
  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

Personalised recommendations