Twelve stream segments were sampled four times in 1998-1999 (one sampling per season) in the drainage basin of the upper São Francisco River (19 masculine 45'-21 masculine 25'S, 49 masculine 05'-51 masculine 30'W), situated in Serra da Canastra National Park, at altitudes ranging from 1,175 to 1,400 m. The macroalgae survey resulted in 30 species, with a predominance of Cyanophyta (12 species = 40%) and Chlorophyta (11 species = 36.5%) and a lower proportion of Rhodophyta (seven species = 23.5%). Two species, Klebsormidium rivulare (Chlorophyta) and Kyliniella latvica (Rhodophyta), were new records for Brazil. Capsosira sp. and Stigonema sp. (Cyanophyta) and the "Chantransia" stage of Batrachospermum (Rhodophyta) were the most widespread macroalgae, occurring in six sampling sites, whereas 11 species were found at only one site. The proportion of macroalgal morphological types were as follows: mats (33%), free filaments (27%), gelatinous filaments (27%), crusts (7%), tufts (3%), and gelatinous colonies (3%). The flora revealed few species in common (4%-8%) with stream macroalgae from other Brazilian regions. The macroalgal communities proved to have species richness values close to the highest values reported in previous studies. The patterns typical for stream macroalgal communities (patchy distribution and dominance of few species) were also found in this basin. However, the stream variables most influential in macroalgal distribution in this study (rocky substratum, low pH, high COD, water color, and current velocity) were essentially the same that best describe the limnological characteristics of this lotic ecosystem. In addition, this combination of variables differed sharply from results of previous studies in other Brazilian stream ecosystems.