Identification of bacteria isolated from diseased Neungee mushroom, Sarcodon aspratus

As the first step in an investigation of the problem with quality deterioration seen in the Neungee mushroom (Sarcodon aspratus) due to bacterial overgrowth during its storage, an attempt to isolate bacterial strains was made using infected gills of Sarcodon aspratus. Five bacterial strains were isolated; one phototrophic cyanobacterial species and four heterotrophic Gram negative rods. The four heterotrophic bacterial isolates (strains P, S, R, and MK1) were subjected to identification based on biochemical characteristics using the Biolog system, cellular fatty acid analysis using the MIDI system, cytology by scanning microscopy, and 16s rDNA sequence analysis. A slow grower, the P strain (ca. 0.7 microm x 1.5 microm), which forms pink colonies on Tryptic Soy agar (TSA) and glucose minimal salt medium containing thiamine (MT medium), belongs to genus Methylobacterium, and is likely M. radiotolerans. The methanol-utilizing capacity of the P strain was confirmed by growth on methanol-supplemented medium as a sole carbon source. Both the S and R strains (ca. 0.5 microm x 0.8 microm) produced smooth and slightly rough white colonies, respectively, on TSA, MT, and potato dextrose (PD) agar are members of the Burkholderia cepacia complex. Although both strains showed some differences from each other in colony morphology, nitrogen fixation capacity, and denitrification, they were considered to be Burkholderia stabilis because their 16s rDNA sequences showed 99.93% similarity with those of B. stabilis LMG 14294T (NCBI AF 148554). The MK1 strain, a rod-shaped bacterium with a tapered end (ca. 0.6 microm x 1.8 microm), produces a copious mucoid substance on MT and PD agar, but not on TSA. Despite extensive identification studies, the M strain is not currently identifiable, which suggests that it is a novel bacterium.