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Altered physiology, cell structure, and gene expression of Theobroma cacao seedlings subjected to Cu toxicity

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Abstract

Seedlings of Theobroma cacao CCN 51 genotype were grown under greenhouse conditions and exposed to increasing concentrations of Cu (0.005, 1, 2, 4, 8, 16, and 32 mg Cu L−1) in nutrient solution. When doses were equal or higher than 8 mg Cu L−1, after 24 h of treatment application, leaf gas exchange was highly affected and changes in chloroplasts thylakoids of leaf mesophyll cells and plasmolysis of cells from the root cortical region were observed. In addition, cell membranes of roots and leaves were damaged. In leaves, 96 h after treatments started, increases in the percentage of electrolyte leakage through membranes were observed with increases of Cu in the nutrient solution. Moreover, there was an increase in the concentration of thiobarbituric acid-reactive substances in roots due to lipid peroxidation of membranes. Chemical analysis showed that increases in Cu concentrations in vegetative organs of T. cacao increased with the increase of the metal in the nutrient solution, but there was a greater accumulation of Cu in roots than in shoots. The excess of Cu interfered in the levels of Mn, Zn, Fe, Mg, K, and Ca in different organs of T. cacao. Analysis of gene expression via RTq-PCR showed increased levels of MT2b, SODCyt, and PER-1 expression in roots and of MT2b, PSBA, PSBO, SODCyt, and SODChI in leaves. Hence, it was concluded that Cu in nutrient solution at doses equal or above 8 mg L−1 significantly affected leaf gas exchange, cell ultrastructure, and transport of mineral nutrients in seedlings of this T. cacao genotype.

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Acknowledgments

We thank Mr. Alberto José dos Santos Júnior and Ms. Valéria Ferreira Fernandes of the Electron Microscopy Center-UESC for the technical assistance on EM. V. L. Souza was supported by FAPESB (Fundação de Amparo a Pesquisa do Estado da Bahia). This work was funded by a grant from USDA, within an international cooperative agreement with CEPLAC, UESC, FUNPAB, and MARS Cacau Inc.

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Correspondence to Alex-Alan F. de Almeida.

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Responsible editor: Philippe Garrigues

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Souza, V.L., de Almeida, A.F., de S. Souza, J. et al. Altered physiology, cell structure, and gene expression of Theobroma cacao seedlings subjected to Cu toxicity. Environ Sci Pollut Res 21, 1217–1230 (2014). https://doi.org/10.1007/s11356-013-1983-4

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Keywords

  • Cacao
  • Heavy metal
  • Lipid peroxidation
  • Photosynthesis
  • Ultrastructure
  • Gene expression