Insights into the saliva of the brown marmorated stink bug Halyomorpha halys (Hemiptera: Pentatomidae)

We examined the salivary gland structure of the brown marmorated stink bug (Pentatomidae: Halyomorpha halys) and developed methods for independent collection of watery saliva and sheath saliva. This stink bug has become a serious invasive pest of agriculture in the United States and its saliva is largely responsible for the damage it causes. We determined by protein gel analysis and shotgun proteomics that the suite of proteins comprising the sheath and watery saliva are very distinct. Our results indicate that a substantial amount of sheath proteins are derived from tomato when stink bugs feed on tomato fruit. Consequently, the sheath saliva is comprised of both insect and plant-derived proteins. Both sheath and watery saliva possessed amylase activities, but polyphenol oxidase and glucose oxidase activities were not detected in either saliva. Peroxidase activity was only detected in salivary sheaths, but only when stink bugs fed on tomato. Proteomic analysis indicated that the peroxidase was likely of plant origin. We also determined that sheath saliva, but not watery saliva elicited the jasmonate inducible defense gene proteinase inhibitor 2 (Pin2), but this induction was only observed when sheaths had been collected from tomato. This indicates that the eliciting factor of the saliva is likely of plant origin. Lastly, neither watery or sheath saliva affected the expression of the salicylate inducible gene pathogenesis related gene (Pr1a-P4).