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.