Indigenous land use and climate have shaped fire regimes in southeast Australia during the Holocene, although their relative influence remains unclear. The archaeologically attested mid-Holocene decline in land-use intensity on the Furneaux Group islands (FGI) relative to mainland Tasmanian and SE Australia presents a natural experiment to identify the roles of climate and anthropogenic land use. We reconstruct two key facets of regional fire regimes, biomass (vegetation) burned (BB) and recurrence rate of fire episodes (RRFE), by using total charcoal influx and charcoal peaks in palaeoecological records, respectively. Our results suggest climate-driven biomass accumulation and dryness-controlled BB across southeast Australia during the Holocene. Insights from the FGI suggest people elevated the recurrence rate of fire episodes through frequent cultural burning during the early Holocene and reduction in recurrent Indigenous cultural burning during the mid–late Holocene led to increases in BB. These results provide long-term evidence of the effectiveness of Indigenous cultural burning in reducing biomass burned and may be effective in stabilizing fire regimes in flammable landscapes in the future.