Diversity of Yeasts and Molds by Culture-Dependent and Culture-Independent Methods for Mycobiome Surveillance of Traditionally Prepared Dried Starters for the Production of Indian Alcoholic Beverages

Abstract

<i>Marcha, thiat, dawdim, hamei, humao, khekhrii, chowan</i>, and <i>phut</i> are traditionally prepared dried starters used for production of various ethnic alcoholic beverages in North East states of India. The surveillance of mycobiome associated with these starters have been revealed by culture-dependent methods using phenotypic and molecular tools. We identified <i>Wickerhamomyces anomalus, Pichia anomala, Saccharomycopsis fibuligera, Pichia terricola, Pichia kudriavzevii</i>, and <i>Candida glabrata</i> by ITS-PCR. The diversity of yeasts and molds in all 40 samples was also investigated by culture-independent method using PCR-DGGE analysis. The average distributions of yeasts showed <i>Saccharomyces cerevisiae</i> (16.5%), <i>Saccharomycopsis fibuligera</i> (15.3%), <i>Wickerhamomyces anomalus</i> (11.3%), <i>S. malanga</i> (11.7%), <i>Kluyveromyces marxianus</i> (5.3%), <i>Meyerozyma</i> sp. (2.7%), <i>Candida glabrata</i> (2.7%), and many strains below 2%. About 12 strains of molds were also identified based on PCR-DGGE analysis which included <i>Aspergillus penicillioides</i> (5.0%), <i>Rhizopus oryzae</i> (3.3%), and sub-phylum: <i>Mucoromycotina</i> (2.1%). Different techniques used in this paper revealed the diversity and differences of mycobiome species in starter cultures of India which may be referred as baseline data for further research.