Inactivation of the Catalytic Subunit of cAMP-Dependent Protein Kinase A Causes Delayed Appressorium Formation and Reduced Pathogenicity of Colletotrichum gloeosporioides

Figure 3

Schematic presentation of CgPKAC gene disruption and DNA blot analysis of CgPKAC gene replacement. (a) Predicted restriction map of the CgPKAC locus in the C. gloeosporioides genome. (b) Gene replacement vector pN-CPKA. The dotted line between (a) and (b) represents the crossing over between two homologous regions that may occur during CgPKAC gene disruption. (c) Predicted restriction map of the Cgpkac deleted allele. (d) Genomic DNA was digested with Xho1 and probed with a 695 bp of 5′CgPKAC fragment. (e) Genomic DNA was digested with Xho1 and probed with a 1.1 kb of hph fragment. In the wild type, a single 3.5 kb DNA fragment was detected whilst for the transformants Cgpkac1 and Cgpkac2, a 5.8 kb band was identified when probed with CgPKAC. Only the Cgpkac1 and Cgpkac2 transformants showed positive signals when probed with the hph gene.