Transformation of maize with genes encoding for insecticidal crystal (Cry) proteins from Bacillus thuringiensis (Bt) could have an impact on the saprophytic survival of plant pathogens and their antagonists on crop residues. We assessed potential effects on the mycotoxin deoxynivalenol (DON)-producing wheat and maize pathogen Fusarium graminearum and on the biocontrol agent Trichoderma atroviride. Purified Cry1Ab protein caused no growth inhibition of these fungi on agar plates. Cry1Ab concentrations above levels common in Bt maize tissue stimulated the growth of F. graminearum. The fungi were also grown on -radiation-sterilized leaf tissue of four Bt maize hybrids and their non transgenic isolines collected at maize maturity on a field trial in 2002 and 2003. Both fungi degraded the Cry1Ab protein in Bt maize tissue. Fungal biomass quantification with microsatellite-based polymerase chain reaction (PCR) assays revealed differential fungal growth on leaf tissue of different maize varieties but no consistent difference between corresponding Bt and non-Bt hybrids. Generally, year of maize tissue collection had a greater impact on biomass production than cultivar or Bt transformation. The mycotoxin DON levels observed in maize tissue experiments corresponded with patterns in F. graminearum biomass, indicating that Bt transformation has no impact on DON production. In addition to bioassays, maize leaf tissue was analyzed with a mass spectrometer?based electronic nose, generating fingerprints of volatile organic compounds. Chemical fingerprints of corresponding Bt and non-Bt leaf tissues differed only for those hybrid pairs that caused differential fungal biomass production in the bioassays. Our results suggest that Cry1Ab protein in maize residues has no direct effect on F. graminearum and T. atroviride but some corresponding Bt/non-Bt maize hybrids differ more in composition than Cry protein content alone, which can affect the saprophytic growth of fungi on crop residues.