Identification of yeast species from orange fruit and juice by RFLP and sequence analysis of the 5.8S rRNA gene and the two internal transcribed spacers
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Las Heras Vázquez, Francisco Javier; Mingorance Cazorla, Lydia; Clemente Jiménez, Josefa María; Rodríguez Vico, FelipeFecha
2003Resumen
Yeast isolates from orange fruit and juice in a spontaneous fermentation were identified and classified by two molecular techniques. The
first was analysis of the restriction pattern generated from the polymerase chain reaction (PCR)-amplified 5.8S rRNA gene and the two
internal transcribed spacers (ITS) using specific primers. The second technique was sequence analysis of the ITS regions using the same
two primers. Nine different restriction profiles were obtained from the size of the PCR products and the restriction analyses with three
endonucleases (CfoI, HaeIII and HinfI). These groups were identified as Candida tropicalis, Clavispora lusitaniae, Hanseniaspora uvarum,
Pichia anomala, Pichia fermentans, Rhodotorula mucilaginosa, Saccharomyces cerevisiae, Saccharomyces unisporus, and Trichosporon asahii. Checking against identification according to morphological, physiological and biochemical traits corroborated this molecular
identification. A total concordance was found in th...
Palabra/s clave
Yeast identication
Restriction fragment length polymorphism-polymerase chain reaction analysis
Sequence analysis
5.8S internal transcribed spacer region