Rapid identification of pickle yeasts by fluorescent PCR and microtemperature-gradient gel electrophoresis.
Monitoring the yeast populations within pickle soaking fluid is imperative for ensuring optimum taste, but these analyses have proven time-consuming and expensive, limiting their industrial application. Here, yeasts were identified in the soaking fluid from Japanese radish pickles using fluorescent PCR amplification of the variable D1/D2 region of the 26S rDNA, followed by analysis with microtemperature-gradient gel electrophoresis (micro-TGGE). This smaller version of the normal TGGE apparatus is capable of analyzing samples 10- to 20-fold faster without sacrificing data quality. Each primer set was labeled with a different fluorescent dye, allowing easy isolation of the various PCR products and identification of the bands corresponding to the various yeasts. The results indicate that fluorescent PCR and micro-TGGE may be a useful new method for rapid, easy monitoring of yeast flora in various food industries. This new method can be used on a daily basis to provide overviews of yeast flora during pickle production, allowing producers to quickly grasp pickle readiness at a single glance.
Tominaga
Tatsuya
T
Saitama Industrial Technology Center North Institute, 2-133, Suehiro, Kumagayashi, Saitama 360-0031, Japan. tominaga@itc-n.pref.saitama.jp
D017931:DNA Primers; D004271:DNA, Fungal / Q000032:analysis / Q000302:isolation & purification; D004275:DNA, Ribosomal / Q000032:analysis / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D005453:Fluorescence; D005456:Fluorescent Dyes; D005516:Food Microbiology; D020459:Genes, rRNA; D016133:Polymerase Chain Reaction / Q000379:methods*; D012335:RNA, Ribosomal / Q000235:genetics; D031224:Raphanus / Q000382:microbiology*; D013696:Temperature; D015003:Yeasts / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*
D017931:DNA Primers; D004271:DNA, Fungal; D004275:DNA, Ribosomal; D005456:Fluorescent Dyes; D012335:RNA, Ribosomal; C065096:RNA, ribosomal, 26S