In the present study, we have investigated the occurrence of yeast flora on several agricultural products coming from crop-growing environments in Cameroon, to provide better knowledge of the biodiversity of yeast flora, and to thus define the impact of this biodiversity on food products. The yeast biodiversity was investigated using traditional culture-dependent methods, along with culture-independent methods. The culture-dependent approach was carried out using both direct and enrichment procedures, to detect the broadest possible presence of yeast species. A total of 151 strains belonging to 26 different yeast species were isolated and identified using restriction pattern analysis of the internal transcribed spacer region 5.8S-ITS and sequence analysis of D1/D2 domain of 26S rRNA gene. The enrichment isolation procedures carried out in high-sugar media allowed the recognition of fermentative species such as Saccharomyces cerevisiae and Torulaspora delbrueckii, which have previously not been detected using direct isolation methodology. The results of culture-independent method using DGGE patterns and sequencing of the DNA bands revealed a lower number of yeast species when compared with the culture-dependent methodology even if the identification of several yeast species not detected by traditional microbiological procedures such as Candida tropicalis and Hanseniaspora uvarum is allowed. Thus, these multiphasic approaches to study yeast biodiversity (culture-dependent and -independent methods) have allowed us to get a more complete picture of the microbial diversity in these natural environments.