Manual annotation of detoxification gene families
We manually annotated five detoxification gene families of cytochrome P450 monooxygenase (P450s), glutathione S-transferase (GSTs), carboxyl/cholinesterases (CCEs), UDP-glycosyltransferases (UGTs) and ATP-binding cassette (ABC) transporters. We used the bioinformatic pipeline BITACORA (Vizueta, Sánchez-Gracia, & Rozas, 2019) to conduct HMMER v3.3 (Finn, Clements, & Eddy, 2011)and BLAST v2.2.31 (Altschul et al., 1990) analyses under a full mode. Hits were filtered with a default cut-off E-value of 10e-5. The HMMs of P450 were downloaded from Pfam v32.0 (Sara El-Gebali et al., 2018), while other HMMs of detoxification gene families were created by HMMER v3.3 (Finn et al., 2011). Orthologs from Bombyx mori and D. melanogaster were used as evidence. The annotated genes were further filtered manually based on gene length and the presence of conserved domains. Genes with a length shorter than 80 amino acids were removed. Orthologs were aligned with the G-INS-I algorithm implemented in MAFFT v7.450 (Katoh & Standley, 2013). A neighbor-joining tree was constructed for each gene family using MEGA7 (Kumar et al., 2016) with 500 bootstrap replicates.