Use of antibiotics to control endophytic bacterial growth migration onto culture medium in Eucalyptus cloeziana F.Muell.: a micropropagation approach

  • Gabriela Ferraz LeoneEmail author
  • Pedro Avelino Maia Andrade
  • Carolina Vieira de Almeida
  • Cristina Vieira de Almeida
  • Fernando Dini Andreote
  • Marcílio de Almeida
Plant Tissue Culture


The natural endophytic bacterial growth migration onto a culture medium is commonly associated with unnecessary microplant discards. In micropropagation procedures, some bacteria can exude from the internal tissues of plants to colonize the culture medium and compete with the plants for nutrients, which may lead to a reduction in plant development. To find an efficient antibiotic protocol to control this bacterial growth migration onto the culture medium without affecting plant development, Eucalyptus cloeziana F.Muell. microstumps were subjected to four antibiotic treatments for 30 d. They were treated with gentamicin, ciprofloxacin, rifampicin, or Timentin®, in addition to the control treatment (antibiotic free). The effects of the antibiotics were monitored weekly, and the endophytic bacterial community structures were evaluated in two periods of plant development (15 d and 30 d).The denaturing gradient gel electrophoresis (DGGE) technique was used to compare the control and post-antibiotic treatment plant microbiological composition, to determine if the antibiotic treatment played a specific role on the endophytic bacterial community structure. The gentamicin treatment was composed of a distinct community from the control treatment. Nonetheless, the plants treated with ciprofloxacin and rifampicin manifested similar endophytic community structures compared to the control. In contrast, plants treated with Timentin® showed a specific bacterial community composition and a higher plant dry mass, number of shoots, and nutritional content. These results suggested that Timentin® treatment could be applied for 30 days to control endophytic bacterial growth migration onto the culture medium, without affecting the homeostatic balance between the bacteria and plants.


In vitro culture Microplants Endophytes Antibiotic therapy Eucalyptus 


Funding Information

The São Paulo Research Foundation (FAPESP; Process No. 2011/15861-6) supported this research.


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Copyright information

© The Society for In Vitro Biology 2019

Authors and Affiliations

  • Gabriela Ferraz Leone
    • 1
    Email author
  • Pedro Avelino Maia Andrade
    • 2
  • Carolina Vieira de Almeida
    • 3
  • Cristina Vieira de Almeida
    • 4
  • Fernando Dini Andreote
    • 2
  • Marcílio de Almeida
    • 1
  1. 1.Department of Biological Science, “Luiz de Queiroz” College of Agriculture (ESALQ)University of Sao Paulo (USP)PiracicabaBrazil
  2. 2.Department of Soil Science, “Luiz de Queiroz” College of Agriculture (ESALQ)University of Sao Paulo (USP)PiracicabaBrazil
  3. 3.Dipartimento di Medicina Sperimentale e ClinicaUniversità Degli Studio FirenzeFlorenceItaly
  4. 4.InVitroPalm (Consulting, Study and Biological Development Ltda.)PiracicabaBrazil

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