The major objective of this study was to describe the effect of water activity and temperature on radial growth and production of ochratoxin A (OTA) and zearalenone (ZEA) on sorghum grains of three Aspergillus tubingensis and three Fusarium incarnatum isolates. The water activity range was 0.91–0.99 aw for F. incarnatum isolates and 0.88–0.99 aw for A. tubingensis isolates. Temperatures of incubation were 15, 25 and 37 °C for both species. Mycotoxin production was determined after 7, 14, 21 and 28 days depending on the growth rate of the six isolates. Maximum growth rates (mm/day) were observed at 37 °C and 0.99 aw for A. tubingensis isolates and at 0.99 aw and 25 °C for F. incarnatum isolates. A. tubingensis was able to grow at 15 °C only at the highest aw levels (0.97 and 0.99 aw). However, at this temperature F. incarnatum grew at 0.94 aw. Optimum ochratoxin A production was observed at 0.97 aw × 37 °Cwhereas optimal conditions for ZEA production varied fromone isolate to another. Moreover, isolates of F. incarnatum from Tunisia do not require high aw and temperature levels to yield maximum levels of ZEA. In general, our results showed that there is no correlation between the growth and production of ZEA in the case of F. incarnatum. This is the first study on the water activity and temperature effect on growth rate and ZEA production of F. incarnatum. Our results show that sorghum grains not only support growth but also OTA and ZEA production by A. tubingensis and F. incarnatum, respectively.