Mandelonitrile produced by commensal bacteria protects the Colorado potato beetle against predation

Larvae and adults of the Colorado potato beetle (Leptinotarsa decemlineata), a major pest of potato crops, display conspicuous coloration to advertise their toxicity to predators. However, the identity of the toxic compounds remains unclear. Here, we show that larvae and adults release toxic hydrogen cyanide (HCN) from the degradation of mandelonitrile and other cyano-compounds, which are produced by commensal bacteria. We isolate the bacterium Proteus vulgaris Ld01 from the insect's gut, and show that it produces HCN and a mandelonitrile-producing cyanoglucoside, amygdalin. Knockout of a gene (hcnB) encoding putative hydrogen cyanide synthase impairs HCN production in P. vulgaris Ld01. Antibiotic treatment of larvae, to eliminate their commensal bacteria, leads to a substantial reduction of HCN emission in larvae and adults. HCN release by bacteria-deprived beetles can be restored by addition of mandelonitrile or by re-infection with P. vulgaris Ld01 (but not with its ∆hcnB1 or ∆hcnB2 mutants). Finally, we use dual-choice experiments to show that domestic chicks prefer to eat bacteria-deprived larvae over control larvae, larvae re-colonized with P. vulgaris Ld01, or mandelonitrile-injected larvae. Our work highlights the role of the beetle's intestinal bacteria in the production of the cyanoglucoside amygdalin and its derived metabolites, including mandelonitrile and HCN, which protect the insect from predation.