Mycobiome Diversity in Traditionally Prepared Starters for Alcoholic Beverages in India by High-Throughput Sequencing Method

Abstract

Chowan, dawdim, humao, hamei, khekhrii, and phut are sun-dried starters used for preparation of alcoholic beverages in North East regions of India. We attempted to profile the mycobiome community in these starters by high-throughput sequencing (HTS) method. All fungal populations were found to be restricted to Ascomycota (67–99%), Zygomycota (0.7–29%), Basidiomycota (0.03–7%), and Chytridiomycota (0.0003%). We found 45 core operational taxonomic units (OTUs) which were universally present and were further weighed to 41 genera level and 22 species level taxonomy. A total number of 594 fungal species were detected by HTS including common species (224), unique species (133) and rare-species (237) in samples of starters. Unique species were recorded in phut (40 species), khekhrii (28), hamei (23), dawdim (21), chowan (13), and humao (8), respectively. Most of the fungal families were found to correlate to a type of nutritional mode and growth morphologies of the community, where saprotrophic mode of mold species were more dominant, whereas morphotypes were more dominant in yeast species.

Diversity of Filamentous Fungi Isolated From Some Amylase and Alcohol-Producing Starters of India

“…Drinking of cereal-based mild to strong alcoholic beverages produced by traditionally prepared amylase and alcoholproducing starters has been a traditional food culture of the ethnic people from the North East states of India for centuries. Traditionally prepared dry starters have consortia of co-existed microbiota containing filamentous molds, yeasts, and bacteria and are crudely sub-cultured through a "backslopping" process by traditional starter-makers Tamang and Sarkar, 1995;Tamang et al, 2007;Sha et al, 2018Sha et al, , 2019, for alcohol production by the Indian people. The pH of traditionally prepared dry starters from India were slightly acidic in nature, perhaps due to accumulation of metabolic organic acids (Ma et al, 2019).…”

“…Some traditionally prepared starters from North East India have been microbiologically analyzed in earlier works and several species of yeasts (Tsuyoshi et al, 2005;Jeyaram et al, 2008Jeyaram et al, , 2011Sha et al, 2017Sha et al, , 2018Sha et al, , 2019 and bacteria (Tamang et al, 2007;Pradhan and Tamang, 2019) were reported. However, detailed taxonomical studies of filamentous molds isolated from traditionally prepared dry starters from North East India have not been reported yet, except for marcha (Tamang et al, 1988;Tamang and Sarkar, 1995;Sha et al, 2017Sha et al, , 2019, thiat (Sha et al, 2017(Sha et al, , 2019, amou, perok-kushi (Das et al, 2017).…”

“…Several species of filamentous molds Yang et al, 2011;Lv et al, 2012a;Chen et al, 2014;Das et al, 2017); yeasts (Hesseltine and Kurtzman, 1990;Tamang and Sarkar, 1995;Tsuyoshi et al, 2005;Jeyaram et al, 2008Jeyaram et al, , 2011Thanh et al, 2008;Fronteras and Bullo, 2017;Sha et al, 2017Sha et al, , 2018Sha et al, , 2019, and bacteria (Hesseltine and Ray, 1988;Tamang et al, 2007;Sha et al, 2017;Roslan et al, 2018) are found to coexist in traditionally prepared dry starters as "micro-resources" which have been sub-cultured to preserve essential microbiota for alcohol production by Asian people for centuries (Tamang et al, 2020). Filamentous fungi present in traditional starters from Asia have several functionalities such as saccharification (Lee and Lee, 2002;Thapa and Tamang, 2004), liquefaction (Suesse et al, 2016), and ethanol production (Dung et al, 2007;Chen et al, 2014) to produce different types of low-alcoholic beverages and highalcoholic distilled liquor.…”

“…Filamentous molds are also responsible for the quality of alcoholic beverages including nutritional values and organoleptic properties such as flavor, taste, and color (Zhang et al, 2015;Tamang et al, 2016). Taxonomical identification of filamentous molds isolated from traditionally prepared dry starters from India have not been reported yet except from marcha (Tamang et al, 1988;Sha et al, 2017Sha et al, , 2019, thiat (Sha et al, 2017(Sha et al, , 2019, amou, and perok-kushi (Das et al, 2017). Mucor circinelloides, Rhizopus chinensis, and Rhizopus stolonifer were reported earlier from marcha samples collected from Nepal, Darjeeling, and Sikkim (Tamang et al, 1988;Tamang and Sarkar, 1995;Thapa and Tamang, 2006;Sha et al, 2017Sha et al, , 2018, Amylomyces rouxii and Rhizopus oryzae from samples of amou, and perok-kushi, traditional starters of Assam (Das et al, 2017).…”

Metagenome gene prediction of naturally fermented milk products of India using PICRUSt2 and Piphillin

“…Targeted gene amplicon sequencing has revolutionized the eld of food microbial ecology [12,13,14,15,16]. High-throughput sequencing methods have been also widely used in many naturally fermented products of North-East India [17,18,19,20]. Naturally fermented milk (NFM) products are popular food items in daily diets of ethnic people of Arunachal Pradesh and Sikkim in India which includes dahi, mohi, gheu, soft-chhurpi, hard-chhurpi, dudh-chhurpi, chhu, somar, maa, philu, shyow, mar, chhurpi/churapi, churkam and churtang/chhurpupu [21].…”