Amplicon Barcoding Based Sequencing Reveals Bacterial Diversity From Himalayan Hotspring of Kotli Azad Jammu and Kashmir

Abstract

Microorganisms are found to exist everywhere including extreme habitats where other forms of life rarely thrive. In this study, culture-independent technique was used to analyze the microbial diversity of water samples from Tata Pani hot spring in district Kotli of Azad Kashmir, Pakistan. Physiochemical analysis was also conducted to study the microbial community composition and diversity of the hot spring in extreme environment. Metagenomic sequencing of five different hot spring samples yielded 875,184 of 16S rRNA sequence reads. The original sequencing output files were submitted in National Centre for Biotechnology Information (NCBI) under the Bioproject ID PRJNA869294 with the accession numbers SRX17083105, SRX17083106, SRX17083107, SRX17083108 and SRX17083109 for 5 hot water samples. Bacterial abundance measures and diversity statistics were assessed using QIIME metagenomics workflow. All the resulting 16S rRNA fragments were then classified into ten bacterial phyla. The highly represented phylum was Proteobacteria (74.68%), followed by Firmicutes (18.58%) and Bacteroidetes (4.25%) respectively. Proteobacteria were detected in a wide temperature range and their relative abundance increased with the decreasing temperature in almost all samples. There were also some unidentified sequences detected at each taxonomic level in all samples. Multivariate analysis indicated that electrical conductivity, temperature and pH are the major environmental factors involved in modelling microbial communities. This is the first study of a high resolution microbial phylogenetic profile of Tatapani hot spring.