Abstract
Marcha, thiat, dawdim, hamei, humao, khekhrii, chowan, and phut 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 Wickerhamomyces anomalus, Pichia anomala, Saccharomycopsis fibuligera, Pichia terricola, Pichia kudriavzevii, and Candida glabrata 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 Saccharomyces cerevisiae (16.5%), Saccharomycopsis fibuligera (15.3%), Wickerhamomyces anomalus (11.3%), S. malanga (11.7%), Kluyveromyces marxianus (5.3%), Meyerozyma sp. (2.7%), Candida glabrata (2.7%), and many strains below 2%. About 12 strains of molds were also identified based on PCR-DGGE analysis which included Aspergillus penicillioides (5.0%), Rhizopus oryzae (3.3%), and sub-phylum: Mucoromycotina (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.
Highlights
Essence of alcoholic fermentation depends on different types of starters that copulate the uniqueness to organoleptic segmentations for ethnic values (Hesseltine, 1983; Steinkraus, 1996; Tamang et al, 2016b)
Ascertaining the cultured diversity, a total of 386 yeasts strains were isolated from 40 samples and characterized by phenotypic assessment on the basis of colony morphology, cell morphology, sugar fermentation, and sugar assimilation tests (Table 2)
By comparing with the yeast database (MicroLog TM System Release 4.2 User Guide 2001, Biolog), the result revealed that maximum identified yeast species were associated with starter having ≥0.75% probability and ≥0.7 similarities index value (Table 3)
Summary
Essence of alcoholic fermentation depends on different types of starters that copulate the uniqueness to organoleptic segmentations for ethnic values (Hesseltine, 1983; Steinkraus, 1996; Tamang et al, 2016b). In Asia, preparation of amylolytic (related to conversion of starch to sugar) and alcoholic (production of alcohol) starter is an innovative back sloping technique of cultivation of native microbiota in the form of dry, flattened, or round balls made up of rice/wheat for production of different traditional alcoholic beverages (Tamang, 2010), locally known as marcha in India, Nepal and Bhutan, benh men in Vietnam, bubod in the Philippines, chiu/chu/daque in China and Taiwan, loogpang in Thailand, ragi in Indonesia, and nuruk in Korea (Tamang, 2016). Few researchers have reported the fungal and bacterial species using both culture-dependent andindependent techniques in some common starter cultures of Asia such as marcha of India (Tsuyoshi et al, 2005; Sha et al, 2017), daqua of China (Wang et al, 2008; Zheng et al, 2012; Lv et al, 2013; Chen et al, 2014; Xu et al, 2017), benh men of Vietnam (Dung et al, 2007; Thanh et al, 2008); nuruk of Korea (Jung et al, 2012), and dombea of Cambodia (Ly et al, 2018)
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