Induction of defense-related genes in tomato plants after treatments with the biocontrol agents Pseudomonas chlororaphis ToZa7 and Clonostachys rosea IK726

  • Nathalie N. KamouEmail author
  • Francisco Cazorla
  • Giannis Kandylas
  • Anastasia L. Lagopodi
Original Paper


Pseudomonas chlororaphis ToZa7 is a promising biocontrol agent possessing valuable characteristics and reducing disease severity caused by Fusarium oxysporum f. sp. radicis-lycopersici (Forl) in tomato. In this study, the strain’s ability to induce three pathogenesis-related (PR) genes (PR-1a, GLUA, and CHI3) in tomato, was studied using quantitative reverse transcription PCR. The genes PR-1a and GLUA were up-regulated after 120 h exposure to P. chororaphis ToZa7 (15.22- and 13.11-fold, respectively), as compared to the untreated control, without challenge inoculation by the pathogen. To study the effects of individual or combined application of P. chororaphis ToZa7 and the compatible biocontrol fungus Clonostachys rosea IK726, challenged with the pathogen, the expression patterns of the above three PR genes were monitored, in tomato roots. Expression of PR1-a was noteworthy, especially 48 h after challenge inoculation, when C. rosea IK726 alone or in combination with P. chororaphis, ToZa7 was pre-inoculated on tomato roots (38.53-fold and 53.74-fold, respectively). Expression of PR1-a, 72 h after challenge inoculation, was the highest in P. chororaphis ToZa7, among biocontrol treatments. Expression of CHI3 was much lower, while up-regulation of GLUA was overall not observed. Confocal laser scanning microscopy of intact tomato roots and bacterial counts of superficially disinfected roots revealed, for the first time, that P. chororaphis ToZa7 colonizes the exterior as well as the internal tissues.


Endophytic colonization Pseudomonas chlororaphis Defense-related proteins Biocontrol Clonostachys rosea 



Authors would like to thank Professor Søren Molin and Dr. Lotte Lambertsen (Technical University of Denmark), and Professor Cayo Ramos (Universidad de Málaga), for kindly supplying the strains and methodology to transform with the mini-Tn7-gfp. Authors are also grateful to professors D.F. Jensen (Swedish University of Agricultural Sciences, Sweden; SLU) and B.J.J. Lugtenberg (Leiden University, The Netherlands for providing strains Clonostachys rosea IK726, and Fusarium oxysporum f. sp. radicis-lycopersici ZUM 2407 (IPO-DLO), respectively. This work was partially supported by Plan Nacional I + D + I from Ministerio de Economía (MINECO; AGL14-52518-C2-1-R), co-financed by FEDER funds (EU). We are also grateful to José A. Gutierrez, Carmen Vida, Sandra Tienda, and David Navas Fernandez for assistance with bacterial transformation and microscopy.

Compliance with ethical standards

Conflict of interest

All authors declare that they have no conflict of interest.

Supplementary material

203_2019_1739_MOESM1_ESM.xlsx (116 kb)
Supplementary file1 (XLSX 117 kb)


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Copyright information

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

Authors and Affiliations

  1. 1.Laboratory of Plant Pathology, School of AgricultureAristotle University of ThessalonikiThessalonikiGreece
  2. 2.Department of MicrobiologyUniversity of MalagaMalagaSpain

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