Rapid differentiation of the closely related Kluyveromyces lactis var. lactis and K. marxianus strains isolated from dairy products using selective media and PCR/RFLP of the rDNA non transcribed spacer 2.
PCR/RFLP of the NTS2 (IGS2) of rDNA was applied to differentiate two closely related yeast species, Kluyveromyces lactis var. lactis (referred to as K. lactis) and K. marxianus. Using specific primers, the NTS2 region was amplified from DNA of both K. lactis and K. marxianus type and collection strains. AluI restriction of amplified fragments generated patterns characteristic for each species. The NTS2 region from K. lactis var. drosophilarum and related species K. aestuarii, K. africanus, K. dobzhanskii, and K. wickerhamii could also be amplified with the same primers, but AluI patterns generated were clearly different. PCR/RFLP of the NTS2 appears thus to be a convenient method for rapid identification of K. lactis and K. marxianus, frequently found in dairy products. This test was validated therefore on K. lactis and K. marxianus from natural habitats. We showed that all yeast strains collected from whey samples and scoring blue on X-gal glucose plates were either K. lactis or K. marxianus. For application purposes, we propose here an approach for quickly screening for K. lactis/marxianus and Saccharomyces cerevisiae in dairy products using X-gal coloured and lysine growth media.
Nguyen
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HV
Laboratoire de Microbiologie et Génétique Moléculaire, INA-PG INRA, Centre Versailles-Grignon, Thiverval-Grignon, France. clib@grignon-inra.fr
Pulvirenti
A
A
Gaillardin
C
C
D000818:Animals; D003470:Culture Media; D004271:DNA, Fungal / Q000235:genetics; D004272:DNA, Mitochondrial / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000235:genetics*; D003611:Dairy Products / Q000382:microbiology*; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D005697:Galactosides / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D007211:Indoles / Q000378:metabolism; D007621:Karyotyping; D007716:Kluyveromyces / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D007785:Lactose / Q000378:metabolism; D008239:Lysine / Q000378:metabolism; D008892:Milk / Q000382:microbiology; D016533:Mycological Typing Techniques; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length
D003470:Culture Media; D004271:DNA, Fungal; D004272:DNA, Mitochondrial; D021903:DNA, Ribosomal Spacer; D005697:Galactosides; D007211:Indoles; C024669:endodeoxyribonuclease AluI; D015252:Deoxyribonucleases, Type II Site-Specific; D005947:Glucose; D007785:Lactose; D008239:Lysine; C044888:5-bromo-4-chloro-3-indolyl beta-galactoside