Co-infection with Wolbachia and Cardinium may promote the synthesis of fat and free amino acids in a small spider, Hylyphantes graminicola.

Associations between endosymbiotic bacteria and their hosts are widespread in nature and have been demonstrated extensively; however, only a few studies have examined how facultative symbionts affect host nutrition and metabolism. To gain insight into the associations between facultative symbionts and host nutrition and metabolic activity, we detected endosymbiotic infection in a small spider species, Hylyphantes graminicola, and established two infectious strains, i.e., W-C+ (Wolbachia negative, Cardinium positive) and W+C+ (Wolbachia positive, Cardinium positive). We then determined the content of fat and free amino acids in W-C+ and W+C+ spiders, respectively. We also detected the transcriptome of H. graminicola and the expression of genes involved in fat and amino acid metabolism at different host ages. Results showed that fat content in W+C+ spiders was higher than that in W-C+ spiders, and free amino acid content was higher in W+C+ males than W-C+ males, with no difference observed in females. Transcriptome analysis identified 144 (W-C+ vs W+C+) differentially expressed genes (DEGs). Moreover, the expression of five genes involved in fat and amino acid metabolism were significantly up-regulated in the third, fourth, and fifth instar stages in W+C+ spiders. This study indicated that Wolbachia and Cardinium co-infection had a pivotal effect on fat and amino acid synthesis in hosts. Moreover, our results provide strong evidence explaining the long-term coexistence of hosts and endosymbionts.