The fungus Trichoderma arundinaceum (Ta37) has a significant biocontrol application which has been related to the production of the trichothecene, harzianum A (2). Previous studies with a strain of T. arundinaceum which was blocked for the production of 2, revealed the existence of a chemical cross-regulation between the biocontrol fungus and its target organism. A study of the secondary metabolome of a single culture of a mutant of T. arundinaceum TaΔTri4 which produces trichothecene biosynthetic intermediates, has now been carried out. The production of secondary metabolites in a co-culture with the phytopathogen, Botrytis cinerea, was then analyzed. The mutant produced a larger quantity of the aspinolides B (6) and C (7) and other derivatives when compared to the wild type Ta37. Ten new metabolites were isolated: three aspinolides 12-14, the γ-lactones 15 and 16, two hemi-ketals 17 and 18 and three aspinolide degradation products, 19, 21 and 22. In the confrontation cultures involving the TaΔTri4 and the B. cinerea B05.10 strains, there was a higher production of the aspinolides B and C by the TaΔTri4 mutant while the production of the botryanes and botcinins by B. cinerea was reduced in the area of interaction between the cultures. These results shed light on the chemical cross-talk and ecological interactions between these fungi.