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Accumulation and distribution of copper in castor bean (Ricinus communis L.) callus cultures: in vitro

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Copper (Cu) accumulation, subcellular distribution and the chemical forms of Cu, and amino acids metabolism were investigated in castor bean (Ricinus communis L.) callus via in vitro culture. The castor bean callus was obtained from the embryo cultured in Murashige and Skoog (MS) medium and then transferred to MS medium with different Cu doses (0, 20, 40, and 60 mg L−1; Cu0, Cu20, etc.) for 28-days cultivation. The stress from Cu inhibited the growth of the castor bean callus, and the Cu content in the castor bean callus increased with the increasing Cu dose, reaching a maximum value of 293.2 mg kg−1 fresh weight (FW) in the 60 mg L−1 Cu treatment. Concentrations of Cu in the cell wall, organelles, and cytoplasm increased significantly with the elevated Cu dose, with the cell wall containing 50.2 % of the total Cu in the 60 mg L−1 Cu treatment. The major Cu fractions were C (bound to the exchangeable polar compounds) (28.1 %) and E (bound to the structural polar compounds and nucleic acids) (27.5 %) in the control treatment, and the main fraction was C (51.2 %) in the treatment with 60 mg L−1 Cu. The concentration of free amino acids in the cytoplasm was closely related to the Cu content in the castor bean callus. In addition, most of the Cu in the cell wall bound with functional groups of the cell chemical components, hydroxyl (–OH), acylamino (–CONH2), and carboxylate ion (–COO). The castor bean exhibited a strong tolerance to Cu, which accumulated mainly in the cell wall.

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1-Naphthaleneacetic acid

MS medium:

Murashige and Skoog medium


Fourier transform infrared spectroscopy


Atomic absorption spectrometry




Tris (hydroxymethyl) aminomethane


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This project was financially supported by National Natural Science Foundation of China (41371470), National High Technology Research and Development Program (863 project) (2012AA101402) and the fundamental research funds for the central universities (2012MBDX005). The authors also want to acknowledge Professor Zdenko Rengel (The University of Western Australia) and Professor Ping Liu (Huazhong Agricultural University) for their help in paper’s languages modification.

Author contributions

Guoyong Huang and Yong Jin designed the experiment and performed all analysis, data interpretation and literature search; Guoyong Huang wrote and revised the manuscript; Wei Kang, Yonghong Liu, and Tao Zou gave some good suggestions on the experiments and participated in the part of experiments; Jin Zheng, Hongqing Hu participated in the experimental design and in the discussion of the manuscript.

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Correspondence to Hongqing Hu.

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Huang, G., Jin, Y., Zheng, J. et al. Accumulation and distribution of copper in castor bean (Ricinus communis L.) callus cultures: in vitro. Plant Cell Tiss Organ Cult 128, 177–186 (2017). https://doi.org/10.1007/s11240-016-1097-z

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  • Castor bean
  • In vitro
  • Callus culture
  • Copper