The tolerance and detoxification of Cr(VI) by eukaryotic microorganisms are of fundamental importance to the treatment of Cr contaminated wastewater. This study presents various significant tolerance and detoxification of hexavalent chromium [Cr(VI)] performed by two strains of PTN7 and PTN10, identified as Trichoderma asperellum with 99% homology. This difference demonstrated by the two similar fungal strains was interpreted based on series Cr(VI) reduction experiments, including those by fungi strain, extracellular polymeric substances (EPS), extracellular and intracellular enzyme in addition to analysis of Cr(VI) and total Cr in the fungal cells. Here, it was confirmed that the higher Cr(VI) tolerance performance by the preferable strain was attributed to stronger extracellular enzyme reduction ability, higher intracellular organic matter absorption capacity and higher intracellular Cr discharge rate from the cell, which could prevent Cr(VI) from getting into the cells, thus reducing potential damage of Cr(VI) to the cells. (C) 2016 Published by Elsevier B.V.