Isolation and Identification of Nitratireductor sp. as a Core LK-Associated Bacterium. LK growing as continuous culture in liquid ASN medium was inoculated in both ASN and LB agar plates. The associated bacteria from LK culture was isolated by using the streak plate method using LB solid media. Genomic DNA was extracted from the isolated bacteria for whole genome sequencing and assembly.

Isolation and Identification of Nitratireductor sp. as a Core LK-Associated Bacterium. Despite detecting multiple bacterial bins in the metagenomic analysis, we successfully isolated a single bacterium growing alongside LK using the streak plate method on LB agar. Whole-genome sequencing of this isolate, followed by genome assembly and taxonomic classification, identified it as Nitratireductor sp. Interestingly, Nitratireductor sp. was not recovered as a metagenomic bin in any of the three sequencing time points. However, mapping the raw metagenomic reads from each time point to the Nitratireductor sp. genome assembly revealed very low representation, with identity percentages of 0.72 in first time point, 0.29 in second time point, and 0.02 in third time point.