Figure 1.

Schematic structure and expression analysis of the metalloprotease MrM35-4. (a) Schematic structuring of the enzyme. The conserved domain and motif are indicated. (b) Transcription of MrM35-4 by the fungus at different stages. RNA was extracted from different samples including: CO, conidia harvested from 2-week-old PDA plate; MY, mycelia harvested from 3-day-old SDB culture; AP, appressoria induced on locust wings for 36 h; HB, hyphal-body cells harvested from the hemocoel of wax moth larvae 48 h post-injection with fungal spores. (c) Protein expression and secretion analysis. The WT spores were inoculated in SDB or MM medium containing 1% silkworm pupa homogenate for 3 days, and the proteins concentrated from the supernatants were used for western blot (WB) analysis with the polyclonal antibody raised against MrM35-4. The uninoculated SDB and 1% cuticle samples were included as mock controls. (d) In vivo detection of the enzyme in insects after injection with fungal spores for different times. The monoclonal antibody raised against the GmAct3 of G. mellonella was used in WB analysis as a reference.