Endophytic bacteria in long-term in vitro cultivated “axenic” pineapple microplants revealed by PCR–DGGE

  • Monita F. Abreu-Tarazi
  • Acácio A. Navarrete
  • Fernando D. Andreote
  • Cristina V. Almeida
  • Siu M. Tsai
  • Marcílio Almeida
Short Communication

Abstract

In vitro propagated plants are believed to be free of microbes. However, after 5 years of in vitro culture of pineapple plants, without evidence of microbial contamination, the use of culture-independent molecular approach [classifying heterogeneous nucleic acids amplified via universal and specific 16S rRNA gene by polymerase chain reaction (PCR)], and further analysis by denaturing gradient gel electrophoresis (DGGE) revealed endophytic bacteria in roots, young and mature leaves of such plants. The amplification of 16S rRNA gene (Bacteria domain) with the exclusion of the plant chloroplast DNA interference, confirmed the presence of bacterial DNA, from endophytic microorganisms within microplant tissues. PCR–DGGE analysis revealed clear differences on bacterial communities depending on plant organ. Group-specific DGGE analyses also indicated differences in the structures of Actinobacteria, Alphaproteobacteria and Betaproteobacteria communities in each part of plants. The results suggest the occurrence of a succession of bacterial communities colonizing actively the microplants organs. This study is the first report that brings together evidences that pineapple microplants, previously considered axenic, harbor an endophytic bacterial community encompassing members of Actinobacteria, Alphaproteobacteria and Betaproteobacteria group which is responsive to differences in organs due to plant development.

Keywords

Micropropagation Ananas comosus Endophyte PCR–DGGE 

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Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  • Monita F. Abreu-Tarazi
    • 1
  • Acácio A. Navarrete
    • 3
  • Fernando D. Andreote
    • 4
  • Cristina V. Almeida
    • 2
  • Siu M. Tsai
    • 3
  • Marcílio Almeida
    • 1
  1. 1.Departamento de Ciências Biológicas, Escola Superior de Agricultura ‘‘Luiz de Queiroz’’Universidade de São PauloPiracicabaBrazil
  2. 2.In Vitro Palm Consultoria E.D.B. Ltda.PiracicabaBrazil
  3. 3.Laboratório de Biologia Celular e Molecular, Centro de Energia Nuclear para AgriculturaUniversidade de São PauloPiracicabaBrazil
  4. 4.Laboratório de Microbiologia Ambiental—Embrapa Meio AmbienteJaguariúnaBrazil

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