Our previous report demonstrated successful isolation of soil-borne bacteria that suppressed the potential of Rhizoctonia solani AG2-2 (IV) causing turfgrass large patch disease when applied to Korean lawngrass (Zoysia japonica). The current study aimed to uncover the mechanisms of this antagonism of Rhizoctonia solani and to define culture conditions for the isolated microbes. We found that two Bacillus isolates, I-009 and FRIN-001-1 strains, produced cellulase and siderophore, but not chitinase, while the Pseudomonas YPIN-022 strain was found to release only siderophore, implying that three antagonistic bacteria commonly interrupt Fe uptake by the large patch pathogen. The I-009 and FRIN-001-1 isolates grew best at 35 and 30°C in growth medium of pH 5 to 8 for 32 and 28 h, respectively, while optimum growth for the YPIN-022 strain was found at 35°C at pH 5 to 9 for 24 h. Good growth of I-009 and YPIN-022 over 24 h was obtained in M9 minimal medium supplemented with 1% sucrose, 0.5% yeast extract and 0.1% potassium chloride. FRIN-001-1 grew well in M9 medium with 1% mannitol, 0.5% yeast extract and 0.1% potassium phosphate dibasic.

Abstract.A large patch disease caused by Rhizoctonia solani AG2-2 (IV) is a serious problem in Korean lawngrass (Zoysia japonica) sites including golf courses and sports fields in Korea. Antagonistic microorganisms against R. solani AG2-2 (IV) were isolated from various forest and crop soil sources in Southern Korea. Among the 61 isolates, I-009, FRIN-001-1, and YPIN-022 strains showing dramatic inhibition of the mycelial growth of R. solani AG2-2 (IV) in the pairing culture were selected as the most potential antagonistic microorganisms for this study. Based on the 16s RNA sequence comparison, I-009 and FRIN-001-1 isolates were identified as Bacillus spp., while YPIN-022 isolate belongs to the genus Pseudomonas. The greater inhibition (clear) zone between two edges of the selected and pathogenic microbes ranged from 11 to 15 mm in three selections, but the others averaged to 7 mm out of 30 mm distance. In another antifungal test using culture filtrate, those three isolates represented a range of 51.7 to 63.5% suppression potential. The selected isolates also inhibited significantly the stem-segment colonization by R. solani AG2-2 (IV) in vivo test by 28.1%, 43.0%, and 23.7% when inoculated with I-009, FRIN-001-1, and YPIN-022, respectively. The highest antagonistic activity for the large patch disease was demonstrated by the isolate FRIN-001-1, which will be useful for developing a bio-pesticide against Rhizoctonia.