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. 2016 Dec 27;114(2):E237–E244. doi: 10.1073/pnas.1611307114

Fig. S4.

Fig. S4.

Targeted gene replacement of M. oryzae CHK1 in the Guy11 background, M. oryzae CDS1 in the Guy11 background, and M. oryzae CDS1 in the Δchk1 background. (A) Southern blot analysis was performed to determine whether targeted gene replacement of CHK1 had taken place, introducing a PvuI restriction digestion, followed by probing with the 1-kb upstream region of CHK1 (Chk1.LF). The result showed a 4.4-kb band, when no gene replacement occurred, and a 2.6-kb band when the corresponding resistance cassette was integrated at the CHK1 locus. The Southern blot confirmed the positive result for T44. (B) Southern blot analysis showing PstI restriction digestion of genomic DNA, followed by probing with the 1-kb upstream region of CDS1 (Cds1.LF). This process generated a 5.5-kb band when no gene replacement had occurred and a 2.5-kb band when the corresponding resistance cassette was integrated at the CDS1 locus. Transformant numbers 2 and 7 correspond to the Δchk1 mutation; number 15 corresponded to an unrelated construct; and numbers 19 and 20 corresponded to the Guy11 genotype. (C) Pathogenicity assays of CO-39 rice leaves inoculated with Guy11, Δchk1, Δcds1, and Δchk1Δcds1 mutants after 5 d postinoculation.