Phylogenetic analysis of insect trehalases based on amino acid sequences. Full-length amino acid sequences were aligned using the Mega 3.1 program to generate a phylogenetic tree (1, class 1 gene of soluble trehalase; 2, class 2 gene of membrane-bound trehalase). A bootstrap analysis was carried out, and the robustness of each cluster was verified in 1000 replications. The scale on the x-axis represents estimated branch lengths and numers indicate bootstrap values. Trehalases were from Aedes aegypti (Aa), Anopheles gambiae (Ag), Apis mellifera (Am), Bombyx mori (Bm), Drosophila melanogaster (Dm), Drosophila simulans (Ds), Homo sapiens (Hs), Mus musculus (Mm), Nasonia vitripennis (Nv), Omphisa fuscidentalis(Of), Pimpla hypochondriaca (Ph), Rattus norvegicus (Rn), Spodoptera exigua (Se), Spodoptera frugiperda (Sf), Tribolium castaneum (Tc) and Tenebrio molitor (Tm). GenBank accession numbers (DNA) are as follows: Aa-2, EAT38444; Ag-2, XP_320471; Am-1, XM_393963; Am-2, XP_394271; Bm-1, BAA13042; Bm-2, AB162717; Dm-2, ABH06695; Ds-2, ABH06710; Hs, NM_007180; Mm, NM_021481; Nv-2, XP_001602179; Of-1, EF426742; Of-2, EF426723; Ph-1, Q8MMG9; Rn, CH473975; Se-1, EU427311; Se-2, EU106080; Sf-1, ABE27189; Tc-1, XP_973919; Tc-2, XP_972610; Tm-1, P32359.