*4.9. Biocontrol Activity of KOF112 against Anthracnose*

Strawberry leaves (*Fragaria ananassa* cv. Hokowase) were detached from seedlings cultivated in a growth chamber. Three detached leaves were sprayed with 500 μL of KOF112 solution (1 × 108 cfu/mL) with a hand sprayer. As the control experiment, sterile water, 10% SCD medium, or *Agrobacterium* sp. isolate CHB3 (1 × <sup>10</sup><sup>8</sup> cfu/mL, containing 10% SCD) was sprayed. After air drying the leaves, the center of each leaf was punctured with a sterile needle. Mycelial disks (6.5 mm diameter) of *C. gloeosporioides* laboratory strain were excised from the leading edge of the colonies on PDA plates, and a disk was placed on the wound of each leaf. After incubation in a moisture chamber at 25 ◦C for 8 d in the dark, disease severity was scored by evaluating the symptom of each leaf as described previously [48]: 0, no symptoms; 1, rot only under the disk; 2, rot two times larger than the disk; 3, rot three times larger than the disk; 4, rot four times larger than the disk; 5, rot more than five times larger than the disk; 6, rot more than six times larger than the disk; 7, rot more than seven times larger than the disk. Four independent experiments were performed with three leaves.
