Figure legends
Figure 1 CRIPSPR-Cas9-based knock out of the GOBPs genes in yellow peach moth. Step1 : positions of each sgRNA and the pair primers for allele-specific PCR detection test. Step2 : Target sequences of the two sgRNAs (blue), the PAM sequences (red) and the cutting sites by the Cas9 protein are indicated with red triangles; a representative chromatogram from PCR products sequencing of individuals after knocking out population with different pair primers.Step3 : Genotyping of individual for deletion of each gene according to banding patterns of the PCR products amplified with a set of the different primer pairs. The band pointed with white arrow is represent homozygous.
Figure 2 Sensitivity test of larvae to artificial diet odorants. Asterisk indicates that significant differences in data (t-test, ***P ≤ 0.001, **P ≤ 0.01 ).
Figure 3 Homozygous strain shows reduced olfactory responses to ecologically relevant single odorants. A heat-map representation of median EAG response values (millivolts) was corrected for the response to the solvent from male and female antennae. Asterisk indicates that significant differences in data when compare with WT (t-test, ***P ≤ 0.001, **P ≤ 0.01, *P ≤ 0.05 ). b redundancy analysis (RDA) on EAG response value in homozygous strain with knocking out GOBP1, GOBP2, and GOBP1/GOBP2. The smaller the angle, the better the correlation. G1, G2, and G1/2 G1 represents the homozygous GOBP1, GOBP2, and GOBP1/2 strain.
Figure 4 Tendency test of adults to apples and sex pheromones. a and b are the tendency of adults to apple, c and d are the tendencies of adults to sex pheromones (ANOVA, ***P ≤ 0.001, 0.01≤ *P ≤ 0.05 ).
Figure 5 STRING Analysis: Protein-protein interaction networks of GOBP1 and GOBP2. a) GOBP1 interacting network with CSPs and OBPs. b) GOBP2 interacting network with CSPs and OBPs. The GOBP1 and 2 had the average local clustering coefficient of 0.839 and 0.861. The insets at the below show the evidence types in STRING network prediction (p-value - <0.05).
Figure 6 SPR of GOBPs-ligands interaction for kinetic measurements. After diluting the ligand at 2 times the concentration, the measured results are represented by different color curves, and the black curve represents the fitting curve; a is GOBP1 reaction with CSP5 and CSP10, respectively; b is GOBP2 reaction with CSP5, CSP10, and OBP17, respectively. All solutions are measured under the condition of pH=7.4.
Figure 7 ITC profiles for the complexation of GOBPs with other ligands. a is GOBP1 reaction with CSP5 and CSP10, respectively; b is GOBP2 reaction with CSP5, CSP10 and OBP17, respectively. All solutions are measured under the condition of pH=7.4.
Figure S1. SDS-PAGE analysis of purified recombinant CSPs and OBPs.
Figure S2. Phylogenetic tree based on amino acid sequences of OBPs and CSPs from Bombyx mori and yellow peach moth. The sequences were aligned using ClustalW. Theevolutionary history was inferred using the neighbor-joining method and MEGA with 1000bootstrap replicates. The OBPs shown in the tree are as follows: Bombyx mori (BmorOBP2, NM_001146714; BmorOBP6, NM_001146718; BmorOBP7, NM_001146719; BmorCSP2, NM_001098308; BmorCSP4, NM_001043587; BmorCSP6, NM_001043935; BmorCSP9, NM_001043604; BmorCSP16, NM_001098312), Conogethes punctiferalis (CpunOBP1, KF026054; CpunOBP2, KF026055; CpunOBP3, KF026056; CpunOBP4, KP985222; CpunOBP5, KP985223; CpunOBP6, KP985224; CpunOBP7, KP985225; CpunOBP8, KP985226; CpunOBP9, KY130463; CpunOBP10, KY130464; CpunOBP11, KY130465; CpunOBP12, KY130466; CpunOBP13, KY130467; CpunOBP14, KY130469; CpunOBP15, KY130470; CpunOBP16, KY130472; CpunOBP17, KY130473; CpunOBP18, KY130474; CpunOBP19, KY130475; CpunOBP20, KY130476; CpunCSP1, KF026049; CpunCSP2, KF026050; CpunCSP3, KY130477; CpunCSP4, KF026057; CpunCSP5, KF026058; CpunCSP6, KF026051; CpunCSP7, KF026052; CpunCSP8, KF026053; CpunCSP9, KY130478; CpunCSP10, KY130479; CpunCSP11, KY130480; CpunCSP12, KY130481; CpunCSP13, KY130482; CpunCSP14, KY130483; CpunCSP15, KY130484).
Figure S3. SPR of GOBPs-ligands interaction for kinetic measurements.