The Crypt-Dwelling Primary Bacterial Symbiont of the Polyphagous Pentatomid PestHalyomorpha halys(Hemiptera: Pentatomidae)

A recent invader of North America, the brown marmorated stink bug (Halyomorpha halys Stål) is a polyphagous pentatomid that harbors a gammaproteobacterial mutualist in the crypts of specialized midgut gastric caeca (region V4). Histological analyses revealed a single rod-shaped morphology abundant in distal V4 midgut caecal crypts. A strong fluorescence signal was detected when thin sections of these tissues were hybridized with a fluorescently-labeled, Enterobacteriaceae-specific oligonucleotide probe. A single operational taxonomic unit (OTU) assigned to the Pantoea genus represented >99% of 3,454 16S rDNA amplicons obtained from midgut V4 tissues and egg samples. Detection of H. halys primary symbiont in DNA extracted from eggs suggested vertical maternal inheritance as the mode of intergenerational transmission. Consistent detection of the bacterial symbiont in geographically distinct H. halys populations strongly supports an intimate association between these two organisms. An inferred phylogeny of gammaproteobacterial symbionts of pentatomids placed the Pantoea-assigned OTU from H. halys within a clade distinct from primary bacterial symbionts of related stink bugs, Nezara viridula (L.) and Eurydema rugosa Motschulsky. Given these data, Candidatus "Pantoea carbekii" is proposed as the name of the primary bacterial symbiont of H. halys.