Red yeast identification by the MSP-PCR technique in a polyphasic approach study

LIBKIND FRATI, D; BRIZZIO, S; RUFFINI, A; GADANHO, M; VAN BROOCK, MR; SAMPAIO, JP

Carlos Paz, Córdoba

Congreso; XXXVIII Reunión Anual de SAIB; 2002

A polyphasic approach study was carried out employing phenotypic and molecular characters for the systematic characterization of 85 red yeast isolates. Samples were taken from Northwestern Patagonian aquatic and terrestrial environments (lakes, soil, nectar, wild fruits, Cyttaria sp. fungus, grapes and grape must). Yeast strains were primary grouped in 4 large groups according to selected physiological tests. Afterwards, yeast isolates were submitted to the Mini/Micro Satellite Primed PCR technique (MSP-PCR) using (GTG)5 and/or M13 primers. DNA banding patterns of the yeast isolates and various Type strains were compared using GelCompar software. Inconclusive results were solved with 26S rDNA (D1/D2 domain) sequence analysis. Sexual compatibility assays and teliospore production tests were applied to detect teleomorphic yeast species. An accurate identification of 90 % of the strains was attained with the MSP-PCR method. The above mentioned techniques combined in a polyphasic approach, led to the identification of 6 genera and 16 red yeast species, including 5 teleomorphic species, and 4 new yeast species. The MSP-PCR fingerprinting based on (GTG)5 or M13 primers generated strain-specific profiles, providing evidence that this method is a simple, reliable and highly reproducible molecular tool for yeast characterization.