Molecular assay to identify Acetobacter diazotrophicus and detect its occurrence in plant tissues (original) (raw)
1998, Canadian Journal of Microbiology
The occurrence of Acetobacter diazotrophicus was directly demonstrated in plant tissues using a species-specific oligonucleotide probe and polymerase chain reaction (PCR) amplification of a 411-bp product. The oligonucleotide probe was derived from the sequence of a highly variable region of 23S rDNA and its specificity was tested with membrane-bound nucleic acids of 112 different microorganisms in hybridization experiments. It was found to be able to discriminate Acetobacter diazotrophicus from other Acetobacter spp. and other reference organisms. PCR amplification from pure cultured cells or colonies showed that the method was sensitive enough to detect as few as 200 cells in the reaction. The presence of Acetobacter diazotrophicus in tissues of micropropagated sugarcane plants inoculated with either this bacterium or a mixture of this bacterium and Herbaspirillum seropedicae was demonstrated by PCR amplification. Acetobacter diazotrophicus could also be detected by the PCR method...
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