FIG 9.

Requirements of the BbAtg3 phosphorylation sites for autophagic flux during B. bassiana development. To determine the necessity of two predicted phosphorylation sites in BbAtg3, the two serine residues (S135 and S136) was mutated into alanine (A) individually and simultaneously. The resultant gene was separately introduced into the A3-GA8 strain, in which autophagy was indicated with GFP-Atg8 fusion protein. Wild-type BbATG3 restored autophagic flux in vacuoles of the ΔBbatg3 strain. Mutation at S135 did not affect the recovery effects of BbATG3. However, mutation of S136, including single and double mutations together with S135, disrupted the recovery effects of BbATG3. “V” indicates the vacuoles in fungal mycelia. Scale bar: 10 μm.