01923naa a2200157 a 450000100080000000500110000800800410001910000180006024500870007826000090016552014460017465000170162070000200163770000210165777300870167816257742004-05-14       2002    bl ---       0--  u    #d1 aSALLES, J. F.  aMolecular method to assess the diversity of Burkholderia species in environmental.  c2002  aIn spite of the importance of many members of the genus Burkholderia in the soil microbial community, no direct method to assess the diversity of this genus has been developed so far. The aim of this work was the development of soil DNA-based PCR-denaturing gradient gel electrophoresis (DGGE), a powerful tool for studying the diversity of microbial communities, for detection and analysis of the Burkholderia diversity in soil samples. Primers specific for the genus Burkholderia were developed based on the 16S rDNA gene sequence and were evaluated in PCRs performed with genomic DNAs from Burkholderia and non-Burkholderia species as the templates. The primer system used exhibited good specificity and sensitivity for the majority of established species of the genus Burkholderia. DGGE analysis of the PCR products obtained showed that there were sufficient differences in migration behavior to distinguish the majority of the 14 Burkholderia species tested. Sequence analysis of amplicons generated with soil DNA exclusively revealed sequences affiliated with sequence of Burkholderia species, demonstrating that the PCR-DGGE method is suitable for studying the diversity of this genus in natural settings. A PCR-DGGE analysis of the Burkholderia communities en two grassland plots revealed differences in diversity mainly between bulk and rhizosphere soil samples; the communities in the latter samples produced more complex patterns.  aBurkholderia1 aSOUZA, F. A. de1 aVAN ELSAS, J. D.  tApplied and Environmental Microbiology, Washingtongv. 68, n.4, p. 1595-1603, 2002