Molecular and culture-based characterization of bacterial community from Manleluag alkaline spring in Pangasinan [Philippines]: Project title:Molecular characterization of bacterial community from Manleluag alkaline spring in Pangasinan

Next generation sequencing technology specifically Illumina HiSeq 2500 platform was employed to obtain the microbial community profile of the Manleluag Hyperalkaline Spring in Pangasinan [Philippines]. Sediment samples was collected from location NO [15 deg 42'11002'N (latitude)/120 deg 16'57.41'E (longitude)]. N1 [15 deg 42'17.30'N (latitude)/120 deg 17'0.70'E (longitude)], and N2 [15 deg 42'17.31'N (latitude)/120 deg 17'0.71'E (longitude). For each location three replicates were collected. DNAs were directly isolated from the sediments. Based on DNA quality and amount, five samples were sequenced, three replicates from NO and two DNA quality and amount, five samples were sequenced, three replicates from NO and two replicates for N2. DNA isolated from N1 is very small in amount and is not of good quality. Correlation analysis and Principal Coordinate Analysis (PCoA) of the obtained data showed that the two replicates of N2 were highly similar. For NO, replicates NO-1 and NO-2 were highly similar but their correlation with NO-3 was a lot lower. Because of this observed difference, the succeeding analyses used NO-1 and NO-2 for location O. Two bioinformatics pipelines include quality assessment of reads, assembly by MEGAHIT and taxonomic annotation using DIAMOND and MEGAN. In one of the pipelines, gene prediction using MetaGeneMark and generation of unigenes using CD-HIT and SoapAligner were employed after the metagenome assembly was generated. The second pipeline directly proceeded to taxonomic annotation after the metagenome assembly. Based on both piplelines, the most abundant phylum is Proteobacteria which includes classes Betaproteobacteria, Alphaproteobacteria, Gammaproteobacteria and Deltaproteobacteria, Acidobacteria, Delinococcus-Thermus, Planctomycetes, Actinobacteria, Armatimonadetes, and Verrumicrobia. Other abundant clases such as Clostridia, Anaerolineae, Bacteroidia, Solibacteres, and Deinococci also belong to the abundant phyla. The most abundant archaea is phylum Euryarcheota with Methanobacterium. The observed predominating taxa reflect the prevalent metabolic pathways that persist in environments characterized by high pH and exhibit serpentinization. The Manleluag Hyperalkaline Spring is a rich source of microorganisms. The data obtained in this study is very good genomic resource that can be used by the future researches.