Expression level heatmap of RNA-Seq, RT-PCR and RT-qPCR results.

<p>Expression data were normalized to ±1 by their range. Values > 0, indicate that the gene has olfactory specificity and is colored red, whereas values < 0, indicate that the gene has non-olfactory specificity and is colored blue. Light and dark colors indicate the ratio of expression level (Antenna/ Non-olfactory organs).</p

Neighbor-joining tree of AquaOBPs.

<p>Values indicated at the nodes are bootstrap values based on 1000 replicates; scale bar = 0.1. Aqua: <i>Ambrostoma quadriimpressum</i>; Tcas: <i>Tribolium castaneum</i>; Dpon: <i>Dendroctonus ponderosae</i>; Malt: <i>Monochamus alternatus</i>; Ldec: <i>Leptinotarsa decemlineata</i>; Ityp: <i>Ips typographus</i>; Tmol: <i>Tenebrio molitor</i>; Acor: <i>Anomala corpulenta</i>. Dval: <i>Dendroctonus valens</i>; Hpar: <i>Holotrichia parallela</i>; Hele: <i>Hylamorpha elegans</i>; Bpra: <i>Brachysternus prasinus</i>; Pjap: <i>Popillia japonica</i>; Eori: <i>Exomala orientalis</i>; Acup: <i>Anomala cuprea</i>; Asch: <i>Anomala schonfeldti</i>; Aoct: <i>Anomala octiescostata</i>. Different colors indicate the types of OBPs.</p

Neighbor-joining tree of AquaORs.

<p>Values indicated at the nodes are bootstrap values based on 1000 replicates; scale bar = 0.1. Aqua: <i>Ambrostoma quadriimpressum</i>; Tcas: <i>Tribolium castaneum</i>; Dpon: <i>Dendroctonus ponderosae</i>; Malt: <i>Monochamus alternatus</i>; Ldec: <i>Leptinotarsa decemlineata</i>; Ityp: <i>Ips typographus</i>; Mcar: <i>Megacyllene caryae</i>; Tmol: <i>Tenebrio molitor</i>; Acor: <i>Anomala corpulenta</i>. Dval: <i>Dendroctonus valens</i>; <i>Agla</i>: <i>Anoplophora glabripennis</i>. Purple indicates the Orco family.</p

Motif analysis of coleopteran CSPs.

<p>Parameters used for motif analysis were minimum width = 6, maximum width = 10, and maximum number of motifs to find = 8. Different colored squares indicate the type and approximate location of each motif on the protein sequence, starting from the N-terminal.</p

Comparison of functional annotations in olfactory and non-olfactory tissue derived transcripts.

<p>A. Comparison of GO annotation functional genes. Red indicates leg, while blue indicates antenna; B. KEGG pathway annotations. The bar represents the number of antenna/leg gene ratios for relevant pathways.</p

Neighbor-joining tree of AquaIRs.

<p>Values indicated at the nodes are bootstrap values based on 1000 replicates, scale bar = 0.1, Aqua: <i>Ambrostoma quadriimpressum</i>; Tcas: <i>Tribolium castaneum</i>; Dpon: <i>Dendroctonus ponderosae</i>; Malt: <i>Monochamus alternatus</i>; Ldec: <i>Leptinotarsa decemlineata</i>; Ityp: <i>Ips typographus</i>; Mcar: <i>Megacyllene caryae</i>; Tmol: <i>Tenebrio molitor</i>; Acor: <i>Anomala corpulenta</i>. Different color ranges indicate different IR families in reference to IR genes of <i>Drosophila melanogaster</i>.</p

RT-qPCR results of candidate olfactory genes of <i>A</i>. <i>quadriimpressum</i>.

<p>RT-qPCR data analysis was performed using the 2^-ΔΔCT method. Error bars indicate the SEM. FA: female antenna; MA: male antenna; FL: female leg; ML: male leg; T: Thorax; W: hind wing. ß-actin was used as an internal control.</p

Motif analysis of coleopteran OBPs.

<p>Parameters used for motif analysis were minimum width = 6, maximum width = 10, and maximum number of motifs to find = 8. Different colored squares indicate the type and approximate location of each motif on the protein sequence, starting from the N-terminal.</p

Neighbor-joining tree of AquaCSPs.

<p>Values indicated at the nodes are bootstrap values based on 1000 replicates; scale bar = 0.1. Aqua: <i>Ambrostoma quadriimpressum</i>; Tcas: <i>Tribolium castaneum</i>; Dpon: <i>Dendroctonus ponderosae</i>; Ityp: <i>Ips typographus</i>; Ldec: <i>Leptinotarsa decemlineata</i>; Tmol: <i>Tenebrio molitor</i>; Acor: <i>Anomala corpulenta</i>; Dval: <i>Dendroctonus valens</i>.</p

RT-PCR results of candidate olfactory genes of <i>A</i>. <i>quadriimpressum</i>.

<p>FA: female antenna; MA: male antenna; FL: female leg; ML: male leg; T: Thorax; W: hind wing. ß-actin was used as an internal control.</p