2.3.1. Effect of co-application of B. linens (IC10) and B. subtilis on disease of tomato fruit
B.linens (IC10) and B. subtilis were transferred from TSA Petri dishes to 15-mL conical tubes containing 5 mL of tryptic soy broth (TSB, Becton Dickinson) using an inoculation loop. The bacteria were grown in an incubator-shaker for 48 h at 26 °C and 175 rpm. Following incubation, bacteria were recovered by centrifugation (5300×g for 20 min). The supernatant was removed and the pellet was washed with 5 mL of sterile distilled water and recentrifuged. The supernatant was discarded and the remaining pellet served as inoculum stocks. The bacterial inocula were adjusted to 5 × 107 cells/mL or 5 × 108 cells/mL with sterile distilled water using a hemacytometer.
A.solani and B. cinerea spores were individually collected by flooding mycelia grown for 1 (A. solani) and 2 weeks (B. cinerea) on PDA with 1 mL sterile distilled water and by gently scraping the surface of the mycelia with a sterile glass rod to dislodge the spores. The spore suspensions were collected by aspiration with a micropipette, placed in a 1.5-mL microcentrifuge tube and diluted to 5 × 104 conidia/mL based on hemacytometer counts of the stock suspension.