Endophytic diazotrophs isolated from molassess grass, Melinis minutiflora,were studied by insert sequence(IS)-PCR fingerprinting patterns and full length of 16S rRNA gene sequences.IS-PCR fingerprinting showed that different patterns appeared between isolates and the type strain DSM 1691 of Azospirillum lipoferum;16S rRNA gene sequence analyses indicated that homology between isolates and known species Azospirillum lipoferum is 97%. These results suggested that isolates from molasses grass are possible a new group of endophytic diazotrophs closely related to Azospirillum lipoferum.