Abstract
Diazotrophic bacteria were isolated from two C4-energy crops, i. e., Miscanthus sinensis and Pennisetum purpureum using semisolid, nitrogen free media. Up to 106 diazotrophic bacteria per gram root fresh weight were determined using the MPNapproach. Among other soil bacteria, Azospirillum and Herbaspirillum isolates were also obtained, which were partially identified as known species. Therefore, the 16S rDNA of representative isolates were sequenced and new specifically binding 16S rRNA binding oligonucleotide probes were developed. Two new species of diazotrophic bacteria, Azospirillum doebereinerae sp. nov., and Herbaspirillum frisingense sp. nov. were identified on the basis of the results of the 16S-rDNA sequences, hybridization with the new probes and physiological tests. Using monospecific polyclonal antibodies, A. doebereinerae ‘GSF71’ was located at the rhizoplane, while H. frisingense ‘Mb11’ was able to enter the roots and to colonize the root interior (intracellular spaces and dead cells in the root cortex and vascular system).
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