Dynamic evolution of the mTHF gene family associated with primary metabolism across life.

BACKGROUND: The folate cycle of one-carbon (C1) metabolism, which plays a central role in the biosynthesis of nucleotides and amino acids, demonstrates the significance of metabolic adaptation. We investigated the evolutionary history of the methylenetetrahydrofolate dehydrogenase (mTHF) gene family, one of the main drivers of the folate cycle, across life. RESULTS: Through comparative genomic and phylogenetic analyses, we found that several lineages of Archaea lacked domains vital for folate cycle function such as the mTHF catalytic and NAD(P)-binding domains of FolD. Within eukaryotes, the mTHF gene family diversified rapidly. For example, several duplications have been observed in lineages including the Amoebozoa, Opisthokonta, and Viridiplantae. In a common ancestor of Opisthokonta, FolD and FTHFS underwent fusion giving rise to the gene MTHFD1, possessing the domains of both genes. CONCLUSIONS: Our evolutionary reconstruction of the mTHF gene family associated with a primary metabolic pathway reveals dynamic evolution, including gene birth-and-death, gene fusion, and potential horizontal gene transfer events and/or amino acid convergence.

Primary Metabolism co-Opted for Defensive Chemical Production in the Carabid Beetle, Harpalus pensylvanicus.

Of the approximately one million described insect species, ground beetles (Coleoptera: Carabidae) have long captivated the attention of evolutionary biologists due to the diversity of defensive compounds they synthesize. Produced using defensive glands in the abdomen, ground beetle chemicals represent over 250 compounds including predator-deterring formic acid, which has evolved as a defensive strategy at least three times across Insecta. Despite being a widespread method of defense, formic acid biosynthesis is poorly understood in insects. Previous studies have suggested that the folate cycle of one-carbon (C1) metabolism, a pathway involved in nucleotide biosynthesis, may play a key role in defensive-grade formic acid production in ants. Here, we report on the defensive gland transcriptome of the formic acid-producing ground beetle Harpalus pensylvanicus. The full suite of genes involved in the folate cycle of C1 metabolism are significantly differentially expressed in the defensive glands of H. pensylvanicus when compared to gene expression profiles in the rest of the body. We also find support for two additional pathways potentially involved in the biosynthesis of defensive-grade formic acid, the kynurenine pathway and the methionine salvage cycle. Additionally, we have found an array of differentially expressed genes in the secretory lobes involved in the biosynthesis and transport of cofactors necessary for formic acid biosynthesis, as well as genes presumably involved in the detoxification of secondary metabolites including formic acid. We also provide insight into the evolution of the predominant gene family involved in the folate cycle (MTHFD) and suggest that high expression of folate cycle genes rather than gene duplication and/or neofunctionalization may be more important for defensive-grade formic acid biosynthesis in H. pensylvanicus. This provides the first evidence in Coleoptera and one of a few examples in Insecta of a primary metabolic process being co-opted for defensive chemical biosynthesis. Our results shed light on potential mechanisms of formic acid biosynthesis in the defensive glands of a ground beetle and provide a foundation for further studies into the evolution of formic acid-based chemical defense strategies in insects.

Pygidial glands of Harpalus pensylvanicus (Coleoptera: Carabidae) contain resilin-rich structures.

The pygidial gland system is a key innovation in adephagan beetles, producing, storing, and spraying defensive chemical compounds. As the source of defensive chemical production and storage, the pygidial gland system experiences severe chemical stress which challenges the integrity of the entire gland system. Here, we utilize autofluorescence-based confocal laser scanning microscopy to examine the morphology of pygidial gland secretory lobes and collecting ductules in a common Pennsylvanian harpaline species, Harpalus pensylvanicus. The glandular units are composed of type-III exocrine cells which empty into resilin-rich ductules, which themselves lead into a larger resilin-rich collecting duct, and ultimately the pygidial reservoir pump. We also utilize histological staining with toluidine blue and brightfield imaging to provide additional support for the presence of resilin in the collecting duct, as toluidine blue has been shown to stain resilin without metachromasia. We hypothesize that the high resilin content of the collecting ducts might be a widespread key evolutionary adaptation to prevent damage caused by physical and chemical stress generated in pump-containing insect exocrine gland systems.

Carabidae Semiochemistry: Current and Future Directions.

Ground beetles (Carabidae) are recognized for their diverse, chemically-mediated defensive behaviors. Produced using a pair of pygidial glands, over 250 chemical constituents have been characterized across the family thus far, many of which are considered allomones. Over the past century, our knowledge of Carabidae exocrine chemistry has increased substantially, yet the role of these defensive compounds in mediating behavior other than repelling predators is largely unknown. It is also unclear whether non-defensive compounds produced by ground beetles mediate conspecific and heterospecific interactions, such as sex-aggregation pheromones or kairomones, respectively. Here we review the current state of non-exocrine Carabidae semiochemistry and behavioral research, discuss the importance of semiochemical research including but not limited to allomones, and describe next-generation methods for elucidating the underlying genetics and evolution of chemically-mediated behavior.