Exploration of Microbial Diversity of Himalayan Glacier Moraine Soil Using 16S Amplicon Sequencing and Phospholipid Fatt
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Exploration of Microbial Diversity of Himalayan Glacier Moraine Soil Using 16S Amplicon Sequencing and Phospholipid Fatty Acid Analysis Approaches Mingma Thundu Sherpa1 · Ishfaq Nabi Najar1 · Sayak Das1 · Nagendra Thakur1 Received: 25 March 2020 / Accepted: 15 October 2020 © Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract Changme Khangpu glacier is located in the northern district of Sikkim which comes under UNESCO heritage site Kanchenjunga Biosphere Reserve which is considered as one of the important biological hotspot regions in the Eastern Himalayas. This is the first report on microbial diversity analysis of moraine soil from one of the unexplored glaciers of Sikkim using high throughput sequencing platform and phospholipid fatty acids analysis (PLFA). It was found that the 16S amplicon sequence comprised 362,902 raw sequences with a sequence length of 150 bp and (G + C) content 52%. A total of 156,821 pre-processed reads were clustered into 378 OTUs (operational taxonomic units) comprising 6 bacterial phyla. The top four dominant phyla based on the 16S amplicon sequences were Proteobacteria (56%), Firmicutes (16%), Actinobacteria (12%), and Bacteroidetes (8%), respectively. PLFA analysis confirmed the dominance of Gram positive bacteria (72%) followed by Gram negative bacteria (32%) and the major fatty acids which are present in the moraine soil sample were PUFA (61%), and 18:2ω6,9c (29%). This is the primary study and first of its kind done on moraine soil from glaciers of Sikkim. Based on 16S amplicon sequencing and PLFA analysis of moraine soil samples from glaciers of Sikkim suggest that this glaciers harbours rich microbial diversity and thus can have wide industrial and biotechnological potential. Thus, there is an escalating scope to further study these extreme biomes with respect to their microbial diversity and their functional capabilities.
Introduction A major portion of the earth’s biosphere is cold (25 NTU), and free chlorine (0.2 ppm). Using google map software and applying geological coordinates the map was created as shown in Supplementary Fig. 1.
40.00% 35.00% 30.00% 25.00% 20.00% 15.00% 10.00% 5.00% 0.00%
Gram Positive
Gram Negative
Anaerobes
Fungi
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M. T. Sherpa et al.
16S Amplicon Sequencing 16S amplicon sequence analysis showed the dominance of phylum Proteobacteria (56%) with the major classes as Beta-Proteobacteria and Gamma-Proteobacteria. After Proteobacteria the phylum Firmicutes (16%) predominant followed by Actinobacteria (12%), and Bacteroidetes (8%) (Fig. 2). The analysis at the genus level showed the dominance of Bifidobacterium, Herminiimonas, Noviherbaspirillum, and Pseudomonas as shown in Fig. 3. Further analysis
at the species level, revealed the presence of 22 bacterial species with dominance of Herminiimonas saxobsidens, Noviherbaspirillum suwonense and unknown species (Fig. 3). The presence of unknown species (8%) strongly suggested that Changme Khangpu glacier moraine harbour the wealth of many novel and uncultivable bacteria. D
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