The 16S rRNA gene sequences registered as GenBank Accession No K

The 16S rRNA gene sequences registered as GenBank Accession No. KC478362 were confirmed by a similarity search of GenBank using the Basic Local Alignment Search Tool (BLAST). The fungal pathogen was cultured on PDA for 7 d, and 5-mm mycelial plugs were placed on the center of the PDA plates. Following this, 10 μL of the bacterial suspension grown ZD1839 in vitro in brain heart infusion (BHI) broth (CONDA, Madrid, Spain) at 28°C for 2 d was spotted 3 cm apart from the mycelial plugs on the media. These agar plates were

incubated at different temperatures of 15°C, 18°C, 21°C, 25°C, and 28°C and the antifungal activity of the bacterial isolates was examined after 1 wk of incubation. SDW was used as an untreated control, and three replications were used for each treatment. The bacterial isolate was cultured in BHI broth at 28°C for 2 d. The bacterial culture was adjusted to concentrations of 106 colony-forming units (CFU)/mL and 108 CFU/mL for treatment. To obtain a cell-free culture filtrate, the bacterial culture was centrifuged at 5,162 g for 20 min and the supernatant was passed through a 0.22 μm Millipore filter (Millipore

Corp., Cork, Ireland). Sterile paper discs (8 mm CP-868596 mouse in diameter) soaked with 40 μL of bacterial suspension or culture filtrate were placed on PDA with approximately 106 conidia/mL plated and incubated at 25°C. After 2 d of incubation, the sizes of clear halos formed around the paper discs were measured to determine the inhibition of conidial germination. To verify the germination rate of conidia, 1 mL of bacterial suspension at low and high concentrations (106 CFU/mL Sclareol and 108 CFU/mL, respectively) was mixed with 1 mL of conidial suspension

containing approximately 106 conidia/mL. Conidial germination was examined at intervals of 6 h and considered positive when the germ-tube length was longer than the nongerminated conidia. Germ-tube lengths were measured randomly up to 100 conidia under a compound light microscope with three replications. The bacterial isolate selected in our study was grown in BHI broth and incubated at 28°C with 200 rpm in a shaking incubator. After incubation for 2 d, bacterial cell suspensions were adjusted to 106 CFU/mL or 108 CFU/mL. Three-yr-old ginseng roots were surface-disinfected with 70% ethanol and 1% sodium hypochlorite for 5 min each and rinsed twice with SDW. These roots were cut into discs of 0.5 cm in thickness and placed on filter paper soaked with SDW in 9-cm petri dishes with three replicates. Cell suspensions (20 μL) were spotted on the ginseng discs. Pure BHI broth was used as a control. Root discs placed on the dishes were incubated at temperatures of 18°C, 21°C, 25°C, and 28°C.

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