Utilizing large Ribosomal Subunit protein L3 for phylogenetic identification of novel bacterial lineages in Zodletone Spring

Abstract

Zodletone Spring is a sulfur and sulfide rich anoxic spring in southwestern Oklahoma characterized by a high level of phylogenetic diversity. We utilized a genome-resolved metagenomics approach to recover genomes of uncultured microbial lineages in the spring. Sediments from the source of the spring were collected, and 551 Gbp of metagenomics raw data were obtained using paired-end Illumina sequencing. The raw data was assembled and genomic bins were recovered from the dataset. A total of 380 genomes with various completion levels were recovered. 321 of these genomes were from the kingdom Bacteria and 59 were from the kingdom Archaea. We used ribosomal protein L3 (RPL3) as a phylogenetic marker to identify genomes belonging to novel lineages within this large dataset. My research focused on the identification of novel class and order level lineages within the phylum Chloroflexi and the classes Deltaproteobacteria (Phylum Proteobacteria), and Clostridia (Phylum Firmicutes). Our analysis putatively identified 19 genomes belonging to 9 novel lineages within the Chloroflexi, 12 genomes defining 8 novel lineages within the Deltaproteobacteria, and 3 genomes belonging to 3 novel lineages within the Clostridia. Future plans include detailed metabolic characterization of these lineages to better understand their metabolic abilities, physiological preferences, and ecological role in Zodletone spring.