Dark septate endophytes (DSEs) constitute a polyphyletic group within the Ascomycota, with global distribution and a wide range of host plant species. The present study evaluated the diversity of DSE in sugarcane roots of the varieties RB867515, RB966928, and RB92579, and four varieties of not commercialized energy cane. A total of 16 DSE strains were isolated, mostly from the varieties RB966928 and RB867515, with six and five isolates, respectively. Just one of the four energy cane varieties had fungi with DSE appearance. The analyses of the DNA sequences from the internal transcribed spacer (ITS) and the large subunit (LSU), in association with the micromorphology of the isolates, allowed the differentiation of the 16 isolates in at least five species, within the families Periconiaceae, Pleosporaceae, Lentitheciaceae, Vibrisseaceae, and Apiosporaceae and the orders Pleosporales, Helotiales, and Xylariales. The order Pleosporales represented 80% of the isolates, and the species Periconia macrospinosa, with six isolates, accounted for the highest isolation frequency. The results confirm the natural occurrence of the DSE symbiosis in sugarcane varieties and the generalist character of these fungi as some of the detected species have already been reported associated with other host plants, ecosystems, and regions of the world
