Combinations of spectrofluorometry and polymerase chain reaction for rapid detection of medium-chain-length polyhydroxyalkanoateaccumulating (Mcl-PHA) bacteria

Abstract

Medium-chain-length polyhydroxyalkanoates (Mcl-PHAs) share physical and material properties that
are considered as an alternative to nonbiodegradable plastics. In this report, a new method was
developed for detection of Mcl-PHA-producing bacteria. The procedure was composed of, first staining
with Nile red and screen under fluorescence microscope at a wavelength of 565 nm, then further
confirmation by amplifying the partial nucleic acid sequence of the PHA synthase gene pha with
polymerase chain reaction (PCR). The stained positive Mcl-PHA-producing bacteria were also detected
while subjected to transmission electron microscopy (TEM). This new method should therefore be
useful for screening Mcl-PHA-producing bacteria from large numbers of microbial cultures timeefficiently.