2016 Volume 15 Issue 4 Pages 103-110
Manganese peroxidase reaction of P. crassa WD1694 cultivated in pulp culture was detected and visualized with tetra-methylbenzidine (TMBZ), a peroxidase substrate for histochemical detection. P. crassa WD1694 was cultivated in fungal media containing unbleached kraft pulp, distilled water and TMBZ. After cultivation for 2 days, flocs including P. crassa WD1694 and pulp were generated in each cultivation condition of solid-state fermentation, an agitated suspension of pulp, and a stationary suspension of pulp. TMBZ staining was observed on the flocs in each culture. Fungal hyphae intimately associated with pulp fibers in the flocs were stained with Coomassie Brilliant Blue, and slime layer covered the fungal hyphae and the pulp in the flocs was visualized with Phloxine B. The TMBZ staining observed in each culture was mainly observed on the hyphal end and along the hyphae in the flocs. Time course analysis of TMBZ and Phloxine B staining revealed that fungal hyphae and slime gathered pulp and made flocs, and the TMBZ staining occurred on the flocs. These results suggest a histological system that MnP reaction occurs on the end of hyphae and along the hyphae in the flocs and that could cause efficient and effective degradation of lignin in pulp.