| Micromelalopha troglodyta（Lepidoptera:Notodontidae）,is one of the defoliator of poplar with wide distribution in China.It had been defined as the third-class hazardous forestry pest by the National Forestry and Grassland Administration in 2019.In this study,we identified the potential olfactory-related genes of M.troglodyta by using RNASeq,phylogenetic tree of the candidate olfactory genes were constructed to exploring the evolutionary relationship of these genes between M.troglodyta and other lepidopteran insects.The expression characteristics of candidate pheromone binding protein genes（Mtro PBPs）were analyzed,and the function of heterologous expressed PBP protein was verified by using fluorescent competitive binding method.The main findings are as follows:（1）53888 Unigenes were obtained from the transcriptome analysis of the adults antennae and larval heads,and there were 20965,13546,15087,20100,15242 and 11934 Unigenes successfully annotated to NR database,Swiss-prot database,Pfam database,Egg NOG database,GO database,and KEGG library,respectively.（2）37 Mtro OBPs（include 3 Mtro PBPs）,23 Mtro CSPs,2 Mtro SNMPs,41 Mtro ORs,5 Mtro GRs,13 Mtro IRs,35 Mtro CXEs were identified in the M.troglodyta transcriptome data by functional annotation and Blast alignment.（3）The tissue expression spectrum of Mtro PBPs showed that Mtro PBP1,2,3 were highly expressed in adults antennae,and the expression level in male antennae was significantly higher than that of females.（4）Mtro PBP1,2,3 showed strong bonding ability to 1-NPN(Mtro PBP1 K1-NPN=8.727μM,Mtro PBP2 K1-NPN=8.185μM,Mtro PBP3 K1-NPN=9.395μM),and Mtro PBPs showed extremely strong bonding ability to M.troglodyta pheromone components（Z13,E15-18:Ald）,Mtro PBP1 Ki=0.68μM,Mtro PBP2 Ki=0.544561284μM,Mtro PBP3 Ki=0.702201992μM.These results lay a foundation for revealing the sex pheromone recognition mechanism of M.troglodyta,and also provide potential targets for the control of M.troglodyta based on reverse chemical ecology. |
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