Bioacoustical data are useful for studying amphibians, especially their conservation, taxonomy, and evolution, among others. Of the 12 species of the Boana polytaenia clade, only B. buriti and B. latistriata have no published information about their advertisement calls. We recorded four males of B. latistriata in its type locality at Parque Nacional do Itatiaia, south-eastern Brazil. We used a Roland R26 digital recorder with a Sennheiser ME-67 microphone and analysed the recordings using the Raven Pro 1.5 software. We recorded two different types of calls (call A and call B). Both were composed of one pulsed note and presented a slightly ascending-descending frequency modulation. Call A was more frequent, having durations between 0.042 and 0.093 s with the dominant frequency ranging from 3375.0 to 3937.5 Hz, and was composed of 11 to 21 pulses separated by intervals that were not fully silent. Call B had durations between 0.711 and 1.610 s, with dominant frequency from 3281.2 to 3750.0 Hz, and was composed of 11 to 29 pulses separated by fully silent intervals. Among the B. polytaenia clade, the calls of B. latistriata are more similar to those of B. bandeirantes, B. beckeri, B. polytaenia, and B. aff. beckeri. The calls of B. latistriata differ from these species in its lower dominant frequency. Boana latistriata present a short, single-note call with a lower pulse period (call A) and a long call with higher pulse period (call B), which differ from the other species of the clade. The coefficients of variation for the various bioacoustical attributes were calculated within- and between-males and these have been discussed. We also report a fight event between two males of B. latistriata. This is the first report of a fight in members of the B. polytaenia clade.