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Engineering expression of bacterial polyphosphate kinase in tobacco for mercury remediation

Abstract

To develop the potential of plants to sequester and accumulate mercurials from the contaminated sites, we engineered a tobacco (Nicotiana tabacum) plant to express a bacterial ppk gene, encoding polyphosphate kinase (PPK), under control of a plant promoter. The designated plant expression plasmid pPKT116 that contains the entire coding region of ppk was used for Agrobacterium-mediated gene transfer into tobacco plants. A large number of independent transgenic tobacco plants were obtained, in some of which the ppk gene was stably integrated in the plant genome and substantially translated to the expected PPK protein in the transgenic tobacco. The presence of Hg2+ did not cause considerable morphological abnormalities in the transgenic tobacco, which grew, flowered, and set seed similarly to the wild-type tobacco on the medium containing normally toxic levels of Hg2+. The ppk-transgenic tobacco showed more resistance to Hg2+ and accumulated more mercury than its wild-type progenitors. These results suggest that ppk-specified polyphosphate has abilities to reduce mercury toxicity, probably via chelation mechanism, and also to accumulate mercury in the transgenic tobacco. Based on the results obtained in the present study, the expression of ppk gene in transgenic tobacco plants might provide a means for phytoremediation of mercury pollution.

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Acknowledgements

We are grateful to Prof. K. Yazaki of Kyoto University for the gift of binary vector pHM6 and A. tumefaciens LBA4404 strain, and for valuable suggestions on the construction of genetically engineered tobacco plants. The wild-type tobacco (N. tabacum cv. Samsun NN) was a generous gift from Japan Tobacco. We also thank C. Ishikawa, R. Isekawa, T. Fuke, H. Hirata, and T. Kawahara of this university for their technical assistance. This work was supported in part by a Grant-in-Aid for Scientific Research (B) (No. 13450216 and 16360267) and Grant-in-Aid for Young Scientists (B) (No. 17790103) from the Ministry of Education, Science, and Culture, Japan.

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Correspondence to Hidemitsu Pan-Hou.

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Nagata, T., Kiyono, M. & Pan-Hou, H. Engineering expression of bacterial polyphosphate kinase in tobacco for mercury remediation. Appl Microbiol Biotechnol 72, 777–782 (2006). https://doi.org/10.1007/s00253-006-0336-3

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Keywords

  • Mercury
  • Transgenic Tobacco
  • Phytoremediation
  • Mercury Resistance
  • Polyphosphate Kinase