Identification of Geotrichum candidum at the species and strain level: proposal for a standardized protocol.
In this study, the M13 primer was used to distinguish Geotrichum candidum from the anamorphic and teleomorphic forms of other arthrospore-forming species (discriminatory power = 0.99). For intraspecific characterization, the GATA4 primer showed the highest level of discrimination for G. candidum among the 20 microsatellite primers tested. A molecular typing protocol (DNA concentration, hybridization temperature and type of PCR machine) was optimized through a series of intra- and interlaboratory trials. This protocol was validated using 75 strains of G. candidum, one strain of G. capitatum and one strain of G. fragrans, and exhibited a discrimination score of 0.87. This method could therefore be used in the agro-food industries to identify and to evaluate biodiversity and trace strains of G. candidum. The results show that the GATA4 primer might be used to differentiate strains according to their ecological niche.
Gente
S
S
Laboratoire de Microbiologie Alimentaire, Université de Caen Basse-Normandie, Esplanade de la Paix, 14032 Caen cedex, France, stephanie.gente@unicaen.fr
Sohier
D
D
Coton
E
E
Duhamel
C
C
Gueguen
M
M
D017104:Bacteriophage M13 / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D004271:DNA, Fungal / Q000032:analysis / Q000592:standards; D020869:Gene Expression Profiling; D005800:Genes, Fungal; D005848:Geotrichum / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D016533:Mycological Typing Techniques / Q000379:methods* / Q000592:standards; D010802:Phylogeny; D017422:Sequence Analysis, DNA
D017931:DNA Primers; D004271:DNA, Fungal