Figure 11.

PiLLP-silenced transformants with reduced catalase genes expression level and lower catalase activities. (A) The expression levels of two catalase genes (PITG_15248 and PITG_07143) in WT, CK, and transformants (S3, S84, O36, and O38). The qRT-PCR assay was performed using cDNAs synthesized from mycelial RNAs of different isolates (WT, CK, STs and OTs). The levels of PITG_15248 or PITG_07143 were determined relative to that of ef1, and the expression level of WT was used as a reference (value, 1.0). (B) In-gel assay of catalase activity. Equal amounts of total protein (20 μg) of different isolates (WT, CK, STs, and OTs) were loaded into Native-PAGE. The gel was immersed in 7 mM H₂O₂ for 10 min after electrophoresis, and then incubated in a 1/1 mixture of freshly prepared 1% potassium hexacyanoferrate (III) and 1% iron (III) chloride hexahydrate. Blue color was observed in the gel except at zones where H₂O₂ was decomposed by catalases. Three replicates were used for each treatment in these tests and the whole experiment was repeated with a different set of biological samples. Bars represent the standard deviation of three replicates. Statistical significance was analyzed using Student's t-test between WT and each transformant (^**P < 0.01).