Understanding the Role of Free-Living Bacteria in the Gut of the Lower Termite Coptotermes gestroi Based on Metagenomic DNA Analysis

SIMPLE SUMMARY: The lower termite Coptotermes gestroi is widely distributed in Northern Vietnam and is an important urban pest that destroys wooden constructions such as pagodas, temples, and furniture. Although the metagenomic DNA of prokaryotes freely living in the termite gut has been sequenced and analyzed, the overall picture of the prokaryotic diversity, including archaea and their function, has not been investigated. The present study, for the first time, revealed the differences in the structure and function of free-living prokaryotes in the termite gut. The bacterial community was formed and adapted to aid the hosts’ survival and development even in the presence of pesticides. Beyond the potential function of the bacteria towards lignocellulose digestion, free-living bacteria and archaea also harbor diverse genes coding for the enzymes/proteins involved in reductive acetogenesis, methanogenesis, methane and sulfur metabolism, and nitrogen fixation and recycling to supply energy for the host; the synthesis of antibiotics for host defense; and the detoxification of aromatic compounds. The present study provides a valuable scientific basis for the mining of novel bacterial genes and the isolation of bacteria from the termite gut for agricultural, environmental, pharmaceutical, and medical applications. ABSTRACT: Termites’ digestive systems, particularly in lower termites with the presence of protozoa, are unique ecological niches that shelter a diverse microbiota with a variety of functions for the host and the environment. In 2012, the metagenomic DNA (5.4 Gb) of the prokaryotes that freely live in the gut of the lower termite Coptotermes gestroi were sequenced. A total of 125,431 genes were predicted and analyzed in order to mine lignocellulolytic genes. however, the overall picture of the structure, diversity, and function of the prokaryotic gut microbiota was not investigated. In the present study, these 125,431 genes were taxonomically classified by MEGAN and functionally annotated by the Kyoto Encyclopedia of Genes and Genomes (KEGG) and by the Carbohydrate-Active enZYmes (CAZy) and HMMER databases. As a result, 95,751 bacterial genes were classified into 35 phyla. The structure of the bacteria, typified by a high ratio of Firmicutes to Bacterioidetes, was distinct from the structure of the entirety of the bacteria in the lower or higher termites’ guts. The archaea (533 genes) were distributed into 4 phyla, 10 classes, 15 orders, 21 families, 47 genera, and 61 species. Although freely living in the guts, the prokaryotic community was formed, developed, and adapted to exhibit unique interactions in order to perform mutual roles of benefit to their hosts. Methanobacteriales, accounting for 61% of the archaea symbionts, seem to play an important role in methanogenesis. Concomitantly, bacterial methanotrophs in the gut utilize methane and combine with other bacterial groups, including potential lignocellulolytic degraders, acetogens, sulfur bacteria, and nitrogen-recycling bacteria, to efficiently convert wood with little nitrogen into acetates via certain pathway modules specified by prokaryotes that freely live in the gut. This forms an important energy source for the termites. Furthermore, bacteria carry 2223 genes involved in the biosynthesis of 17 antibiotic groups. The gut bacteria also possess genes for the degradation of 18 toxic aromatic compounds, of which four are commercial pesticides against termites commonly used for the preservation of wooden constructions. Eight of the eighteen pathways were the first to be reported from the termite gut. Overall, this study sheds light on the roles of the freely living bacteria and archaea in the C. gestroi gut, providing evidence that the gut microbiome acts as the second host genome, contributing both nutrients and immunity to support the host’s existence, growth, and development.