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Microbial Ecology

, Volume 50, Issue 1, pp 128–139 | Cite as

Design, Development, and Use of Molecular Primers and Probes for the Detection of Gluconacetobacter Species in the Pink Sugarcane Mealybug

  • Ingrid H. Franke-Whittle
  • Michael G. O’Shea
  • Graham J. Leonard
  • Lindsay I. Sly
Article

Abstract

Molecular tools for the species-specific detection of Gluconacetobacter sacchari, Gluconacetobacter diazotrophicus, and Gluconacetobacter liquefaciens from the pink sugarcane mealybug (PSMB) Saccharicoccus sacchari Cockerell (Homiptera: Pseudococcidae) were developed and used in polymerase chain reactions (PCR) and in fluorescence in situ hybridizations (FISH) to better understand the microbial diversity and the numerical significance of the acetic acid bacteria in the PSMB microenvironment. The presence of these species in the PSMB occurred over a wide range of sites, but not in all sites in sugarcane-growing areas of Queensland, Australia, and was variable over time. Molecular probes for use in FISH were also designed for the three acetic acid bacterial species, and shown to be specific only for the target species. Use of these probes in FISH of “squashed” whole mealybugs indicated that these acetic acid bacteria species represent only a small proportion of the microbial population of the PSMB. Despite the detection of Glac. sacchari, Glac. diazotrophicus, and Glac. liquefaciens by PCR from different mealybugs isolated at various times and from various sugarcane-growing areas in Queensland, Australia, these bacteria do not appear to be significant commensals in the PSMB environment.

Keywords

Sugarcane Acetic Acid Bacterium EUB338 Probe Gluconacetobacter Diazotrophicus Formamide Concentration 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

This work was funded by the Sugar Research Development Corporation, Australia. We are indebted to staff of the Bureau of Sugar Experiment Stations (BSES) for their assistance in mealybug collection throughout Queensland, and especially to Tony Blatch (BSES, Nambour) for his assistance in numerous field trips for mealybug and sugarcane collection. Thanks are also extended to Mark Fegan (Department of Microbiology and Parasitology) and to Niall Masel (BSES, Indooroopilly) for various useful discussions.

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Copyright information

© Springer Science+Business Media, Inc. 2005

Authors and Affiliations

  • Ingrid H. Franke-Whittle
    • 1
  • Michael G. O’Shea
    • 2
  • Graham J. Leonard
    • 2
  • Lindsay I. Sly
    • 1
  1. 1.Centre for Bacterial Diversity and Identification, Department of Microbiology and ParasitologyThe University of QueenslandBrisbaneAustralia
  2. 2.Bureau of Sugar Experiment StationsBrisbaneAustralia

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