Synergistic composition useful as microbiological growth medium for rapid screening of phosphate accumulating microorganisms

摘要

Screening isolates for phosphate accumulation by quantitative methods requires investment of time, labor and chemicals. There has been paucity of a medium and method for screening phosphate-accumulating microorganisms (PAOs) from the environment. Therefore, a new screening strategy is needed towards isolating effective PAOs in environments with diverse microbial populations and limited organic resources, where it is possible that PAOs are often out-competed by organisms capable of utilizing available nutrients more rapidly. In view of these problems, a medium optimal for an efficient screening of P-accumulating microorganisms was developed, which is rapid and allows many PAOs to be screened. Thus, the present invention utilizes Toluidine Blue-O (TBO) a blue colored dye, which decolorizes due drop in the concentration of phosphate in the medium, as an indicator to quickly evaluate the P-accumulating microorganisms based upon visual observations. Fast decolorization of the dye is an indicator of a superior phosphate accumulating microbe. Therefore, colorimetric reaction using PAM-TBO of the present work was further applied in a novel, extremely high-throughput manner to detect individual microbial isolates, growing on a PAM Petri plate. Adaptation of PAM-TBO assay in screening PAOs in environmental samples for microtitre plate assay enhanced the effectiveness of the assay system efficiently and effectively.