Pheromone-binding proteins (PBPs) have been extensively investigated in lep-idopteran moths,but their evolution and function in hemipteran species remain unclear.Our previous study demonstrated that an odorant-binding protein,OBP4,of the mirid bug Adelphocoris lineolatus functions as a candidate hemipteran PBP but clustered with lepidopteran antennae-binding proteins (ABPs) rather than in the PBP/general odorant-binding protein (GOBP) clade.In this study,we hypothesized that origin and function of PBPs in hemipteran bugs may differ from those of lepidopteran moths.To test this hypoth-esis,we first constructed a phylogenetic tree using insect OBPs from sister hemipteran and holometabolous lineages,and the results indicated that neither OBP4 nor other types of candidate PBPs of mirid bugs clustered with the lepidopteran PBP/GOBP clade.Then,a fluorescence competitive binding assay was employed to determine binding affinities of recombinant OBP4 protein to host plant volatiles,with functional groups different from A.lineolatus sex pheromone components.The results revealed that OBP4 highly bound the female adult attractant 3-hexanone and 15 other mirid bug biologically active plant volatiles.Finally,we examined cellular expression profiles of OBP4 in putative antennal sensilla that are related to female A.lineolatus host plant location.The fluorescence in situ hybridization and immunocytochemical labeling assay showed that the OBP4 gene was highly expressed in the multiporous olfactory sensilla medium-long sensilla basicon-ica rather than in the short sensilla basiconica or uniporous sensilla chaetica.These results,together with those of our previous studies,indicate that OBP4 not only functions in recog-nition of bug-produced sex pheromones in males,but is probably involved in detection of host plant volatiles in both A.lineolatus sexes.Our findings support the hypothesis that the origin and function of PBPs in hemipteran bugs differ from those of well-known PBPs in lepidopteran moths,which provides a novel perspective on evolutionary mechanisms of sex pheromone communication across insect orders.

Qian Wang;Yujie Li;Qi Wang;Liang Sun;Yongjun Zhang

Key Laboratory of Biology,Genetics and Breeding of Special Economic Animals and Plants,Ministry of Agricultura and Rural Affairs,Tea Research Institute,Chinese Academy of Agricultural Sciences,Hangzhou,310008,China;College of Agriculture and Food Science,Zhejiang A & F University,Hangzhou,311300,China;State Key Laboratory for Biology of Plant Diseases and Insect Pests,Institute of Plant Protection,Chinese Academy of Agricultural Sciences,Beijing,100193,China

中国昆虫科学（英文版）

[1]Qian Wang;Yujie Li;Qi Wang;Liang Sun;Yongjun Zhang-.The Adelphocoris lineolatus OBP4:Support for evolutionary and functional divergence of a mirid pheromone-binding protein from that found in lepidopteran moths)[J].中国昆虫科学（英文版）,2022(01):151-161

Adelphocoris,lineolatus,OBP4,mirid,lepidopteran,moths,lep,idopteran,hemipteran,hypoth,holometabolous,hexanone,immunocytochemical,multiporous,basicon,basiconica,uniporous,chaetica

Support,evolutionary,functional,divergence,binding,from,that,found,Pheromone,proteins,PBPs,have,been,extensively,investigated,but,their,species,remain,unclear,Our,previous,study,demonstrated,odorant,functions,candidate,clustered,antennae,ABPs,rather,than,general,GOBP,clade,In,this,hypothesized,origin,bugs,may,those,To,test,first,constructed,phylogenetic,tree,using,insect,OBPs,sister,lineages,results,indicated,neither,nor,other,types,Then,fluorescence,competitive,assay,was,employed,determine,affinities,recombinant,host,plant,volatiles,groups,different,components,revealed,highly,bound,female,adult,attractant,biologically,active,Finally,examined,cellular,expression,profiles,putative,antennal,sensilla,are,related,location,situ,hybridization,labeling,showed,expressed,olfactory,medium,long,short,These,together,our,studies,not,only,recog,nition,produced,pheromones,males,probably,involved,detection,both,sexes,findings,support,hypothesis,well,known,which,provides,novel,perspective,mechanisms,communication,across,orders

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