Endophytic bacteria from Ilex paraguariensis shoot cultures: localization, characterization, and response to isothiazolone biocides

  • Claudia Luna
  • Raúl Acevedo
  • Mónica Collavino
  • Ana González
  • Luis Mroginski
  • Pedro Sansberro
Micropropagation
First Online:
Received:
Accepted:

Abstract

Nodal segment explants of Ilex paraguariensis, collected from greenhouse-grown plants, were found to contain endophytic bacteria. After culturing in bioreactors, 16 rRNA gene analyses and analytical profile index biochemical tests were used to identify these bacteria as Stenotrophomonas malthophilia. The presence of bacterial cells in the intercellular spaces of stem cortical parenchyma was detected in histological sections by scanning electron microscopy. A range of commercial isothiazolone biocides were tested for their ability to repress the growth of Gram-negative bacteria grown in liquid media during the micropropagation phase. The addition of 0.75 ml l−1 Delcide™ TG (5-chloro-2-methyl-4-isothiazolin-3-one + 2-methyl-4-isothiazolin-3-one, 1.05% and 0.45%, respectively) to the culture media resulted in 100% visibly clean cultures, with no suppression of shoot growth.

Keywords

Bioreactors Contamination 16S rDNA analysis Woody plant species Micropropagation 
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Notes

Acknowledgments

The authors are grateful for funding support from SGCyT-UNNE (grant no. 014/10, 005/11), CONICET (PIP 0734), and Establecimiento Las Marías S.A.C.I.F.A. We extend our deepest appreciation to the companies that provided the biocides.

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Copyright information

© The Society for In Vitro Biology 2013

Authors and Affiliations

  • Claudia Luna
    • 1
  • Raúl Acevedo
    • 1
  • Mónica Collavino
    • 1
  • Ana González
    • 1
  • Luis Mroginski
    • 1
  • Pedro Sansberro
    • 1
  1. 1.Instituto de Botánica del NordesteFacultad de Ciencias Agrarias (UNNE)CorrientesArgentina

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