An Ash1-Like Protein MoKMT2H Null Mutant Is Delayed for Conidium Germination and Pathogenesis in Magnaporthe oryzae

Figure 1

The strategy for MoKMT2H gene replacement with PCR. (a) Schematic representation of the genomic DNA of MoKMT2H. Blue boxes indicate exons. The ATG start codon and TAA stop codon are indicated. The EcoRI restriction enzyme sites are indicated. The red line labeled “probe” shows the region used for Southern blot analysis. The upstream and downstream flanking sequences were amplified with primer pairs LBCK/LB-R and RBCK/RB-F, respectively. The fused DNA fragments with HYG were amplified with LB-F/HYG-R1 and HYG-F1/RB-R. The DNA fragments used for transformants were amplified with LB-F/RB-F. (b) Southern blot analysis of EcoRI-digested genomic DNA from wild-type P131 and the neomycin-resistant transformants KO1, KO2, and KO3. Blots were hybridized with probe as indicated in (a). The DNA fragment used for a probe was amplified with primer pair Pr2937F/Pr2937R. (c) The transformants KO2 and KO3 were confirmed by amplification with primer pairs UPF/HYG-R and UPR/HYG-F.

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