Applied Microbiology and Biotechnology

, Volume 104, Issue 3, pp 1319–1330 | Cite as

Use of phage ϕ6 to inactivate Pseudomonas syringae pv. actinidiae in kiwifruit plants: in vitro and ex vivo experiments

  • Larindja A. M. Pinheiro
  • Carla Pereira
  • M. Esther Barreal
  • Pedro Pablo Gallego
  • Victor M. BalcãoEmail author
  • Adelaide AlmeidaEmail author
Environmental biotechnology


Over the last years, the global production and trade of kiwifruit has been severely impacted by Pseudomonas syringae pv. actinidiae (Psa), a phytopathogen that causes a disease in kiwifruit plants known as bacterial canker. The available treatments for this disease are still scarce, with the most common involving frequently spraying the orchards with disinfectants, copper-based bactericides and/or antibiotics. Moreover, these treatments should be avoided due to their high toxicity to the environment and promotion of bacterial resistance. Phage therapy may be an alternative approach to inactivate Psa. The present study investigated the potential application of the already commercially available bacteriophage (or phage) ϕ6 to control Psa infections. The inactivation of Psa was assessed in vitro, using liquid culture medium, and ex vivo, using artificially contaminated kiwifruit leaves with two biovar 3 (a highly aggressive pathogen) strains (Psa CRA-FRU 12.54 and Psa CRA-FRU 14.10). In the in vitro experiments, the phage ϕ6 was effective against both strains (maximum reduction of 2.2 and 1.9 CFU/mL for Psa CRA-FRU 12.54 and Psa CRA-FRU 14.10, respectively). In the ex vivo tests, the decrease was lower (maximum reduction 1.1 log and 1.8 CFU/mL for Psa CRA-FRU 12.54 and Psa CRA-FRU 14.10, respectively). The results of this study suggest that the commercially available phage ϕ6 can be an effective alternative to control Psa infections in kiwifruit orchards.


Phage treatment Phage ϕ6 Pseudomonas syringae pv. actinidiae Kiwifruit canker Sustainable bactericide treatment 



Thanks are also due to the Department of Biology and University of Aveiro where this research work was carried out.

Author contributions

Larindja A. M. Pinheiro, Carla Pereira and Victor M. Balcão performed the experiments. Larindja A. M. Pinheiro, Victor M. Balcão and Carla Pereira prepared the paper. M. Esther Barreal, Pedro Pablo Gallego revised the paper and contributed with Psa strains. Adelaide Almeida supervised the work, prepared and revised the paper and contributed with reagents and analysis tools.

Funding information

FCT/MCTES provided financial support to CESAM (UID/AMB/50017/2019), through national funds. Carla Pereira was supported by a Junior Research contract (CEEC Individual/03974/2017) financed by the Portuguese Foundation for Science and Technology (FCT). Project funding by Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP, São Paulo, Brazil) (BPE fellowship Ref. No. 2018/05522-9, Project PsaPhageKill, granted to Victor M. Balcão) is hereby gratefully acknowledged. This work also received support from CNPq, National Council for Scientific and Technological Development Brazil, in the form of Research Productivity (PQ) fellowships granted to Victor M. Balcão (Refs. No. 306113/2014-7 and 308208/2017-0).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethics statement

This article does not contain any studies with human participants or animals performed by any of the authors.


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Larindja A. M. Pinheiro
    • 1
  • Carla Pereira
    • 1
  • M. Esther Barreal
    • 2
  • Pedro Pablo Gallego
    • 2
  • Victor M. Balcão
    • 1
    • 3
    Email author
  • Adelaide Almeida
    • 1
    Email author
  1. 1.Department of Biology and CESAMUniversity of Aveiro, Campus Universitário de SantiagoAveiroPortugal
  2. 2.Department of Plant Biology and Soil Science, Faculty of BiologyUniversity of VigoVigoSpain
  3. 3.PhageLab – Laboratory of Biofilms and BacteriophagesUniversity of SorocabaSorocabaBrazil

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