article · Microbes and Environments
A specialised culture medium named PSR2A-C/T has been created to isolate Flavobacterium and Chryseobacterium bacteria from plant rhizosphere soil. The medium adapts standard Reasoner's 2A agar by autoclaving phosphate and agar separately and adding specific concentrations of cycloheximide and tobramycin. In tests comparing nine different culture media using Welsh onion rhizosphere samples, Flavobacterium strains grew exclusively on PSR2A-C/T. Additionally, the medium demonstrated a higher recovery rate for Chryseobacterium strains compared to all other tested formulations, including various standard nutrient agars and antibiotic combinations. The formulation was further validated using onion rhizosphere soil, successfully isolating both bacterial groups at rates comparable to those observed in Welsh onion samples. This formulation offers a reliable method for targeting and recovering these specific soil microorganisms.
Rhizosphere bacteria play critical roles in soil health and plant-microbe interactions, yet isolating specific strains from complex soil communities is often difficult. By establishing a targeted growth medium that selectively recovers Flavobacterium and Chryseobacterium, researchers gain a reliable tool to extract, study, and utilise beneficial root-associated bacteria that may otherwise be overgrown or missed using standard laboratory media.
The medium serves as a practical laboratory protocol for researchers and agricultural biotechnology teams seeking to screen and isolate rhizosphere bacteria. In terms of readiness, this is an applied and tested laboratory tool validated across Welsh onion and onion soils. It could be adopted directly by diagnostic and research laboratories, though the abstract focuses strictly on selective bacterial isolation and does not describe downstream product development pathways.
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An effective medium designated phosphate separately autoclaved Reasoner's 2A supplemented with cycloheximide and tobramycin (PSR2A-C/T) has been developed for the isolation of Flavobacterium and Chryseobacterium strains from the plant rhizosphere. It consists of Reasoner's 2A agar (R2A) prepared by autoclaving phosphate and agar separately and supplementing with 50 mg L(-1) cycloheximide and 1 mg L(-1) tobramycin. A comparison was made among the following nine media: PSR2A-C/T, PSR2A-C/T supplemented with NaCl, R2A agar, R2A agar supplemented with cycloheximide and tobramycin, 1/4-strength tryptic soy agar (TSA), 1/10-strength TSA, soil-extract agar, Schaedler anaerobe agar (SAA), and SAA supplemented with gramicidin, for the recovery of Flavobacterium and Chryseobacterium strains from the Welsh onion rhizosphere. Flavobacterium strains were only isolated on PSR2A-C/T, and the recovery rate of Chryseobacterium strains was higher from PSR2A-C/T than from the eight other media. In order to confirm the effectiveness of PSR2A-C/T, bacteria were isolated from onion rhizosphere soil with this medium. Flavobacterium and Chryseobacterium strains were successfully isolated from this sample at a similar rate to that from the Welsh onion rhizosphere.
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DOI: 10.1264/jsme2.me15144
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