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Front Physiol
2017 Jan 01;8:953. doi: 10.3389/fphys.2017.00953.
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Chemosensory Gene Families in Ectropis grisescens and Candidates for Detection of Type-II Sex Pheromones.
Li ZQ
,
Luo ZX
,
Cai XM
,
Bian L
,
Xin ZJ
,
Liu Y
,
Chu B
,
Chen ZM
.
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Tea grey geometrid (Ectropis grisescens), a devastating chewing pest in tea plantations throughout China, produces Type-II pheromone components. Little is known about the genes encoding proteins involved in the perception of Type-II sex pheromone components. To investigate the olfaction genes involved in E. grisescens sex pheromones and plant volatiles perception, we sequenced female and male antennae transcriptomes of E. grisescens. After assembly and annotation, we identified 153 candidate chemoreception genes in E. grisescens, including 40 odorant-binding proteins (OBPs), 30 chemosensory proteins (CSPs), 59 odorant receptors (ORs), and 24 ionotropic receptors (IRs). The results of phylogenetic, qPCR, and mRNA abundance analyses suggested that three candidate pheromone-binding proteins (EgriOBP2, 3, and 25), two candidate general odorant-binding proteins (EgriOBP1 and 29), six pheromone receptors (EgriOR24, 25, 28, 31, 37, and 44), and EgriCSP8 may be involved in the detection of Type-II sex pheromone components. Functional investigation by heterologous expression in Xenopus oocytes revealed that EgriOR31 was robustly tuned to the E. grisescens sex pheromone component (Z,Z,Z)-3,6,9-octadecatriene and weakly to the other sex pheromone component (Z,Z)-3,9-6,7-epoxyoctadecadiene. Our results represent a systematic functional analysis of the molecular mechanism of olfaction perception in E. grisescens with an emphasis on gene encoding proteins involved in perception of Type-II sex pheromones, and provide information that will be relevant to other Lepidoptera species.
Figure 1. Annotation summaries for E. grisescens transcripts. (A) Species distribution of transcripts with best hit annotation terms in non-redundant (NR) database. (B) Gene ontology (GO) classifications of E. grisescens transcripts.
Figure 2. Alignment of amino acid sequences of EgriOBPs and EgriCSPs. (A) Alignment of amino acid sequences of the EgriOBPs. (B) Alignment of amino acid sequences of EgriCSPs. Boxes indicate predicted signal peptides, blue highlight indicates conserved cysteines.
Figure 3. Phylogenetic analysis of EgriOBPs with other typical insect OBPs. Phylogenetic tree was constructed in PhyML3.0 using maximum likelihood method.
Figure 4. Phylogenetic analysis of EgriCSPs with other typical insect CSPs. Phylogenetic tree was constructed in PhyML3.0 using maximum likelihood method.
Figure 5. Phylogenetic analysis of EgriORs with other typical insect ORs. Phylogenetic tree was constructed in PhyML3.0 using maximum likelihood method.
Figure 6. Phylogenetic analysis of EgriIRs with other typical insect IRs. Phylogenetic tree was constructed in PhyML3.0 using maximum likelihood method.
Figure 7. Tissue expression profiles and abundance of selected EgriOBP and EgriCSP genes in antennae based on relative mRNA quantity and RPKM values. Heat map illustrates Log10-transformated mRNA expression levels of EgriOBP and EgriCSP in different tissues. Histogram shows RPKM values of EgriOBP and EgriCSP. (A) Tissue expression profile and RPKM values of selected EgriOBP genes. (B) Tissue expression profile and RPKM values of selected EgriCSP genes. FA, female antennae; MA, male antennae; FH, female head without antennae; MH, male head without antennae; FT, female thorax; MT, male thorax; FAb, female abdomen without pheromone glad; MAb, male abdomen; FL, female legs; ML, male legs; FW, female wings; MW, male wings; FPr, female proboscis; MPr, male proboscis; Pg, pheromone gland. *P-value < 0.05.
Figure 8. Tissue expression profiles and abundance of selected EgriOR and EgriIR genes in antennae based on relative mRNA quantity and RPKM values. Heat map illustrates Log10-transformed mRNA expression levels of EgriOR and EgriIR in different tissues. Histogram shows RPKM values of EgriOR and EgriIR. (A) Tissue expression profile and RPKM values of selected EgriOR genes. (B) Tissue expression profile and RPKM values of selected EgriIR genes. FA, female antennae; MA, male antennae; FH, female head without antennae; MH, male head without antennae; FT, female thorax; MT, male thorax; FAb, female abdomen without pheromone gland; MAb, male abdomen; FL, female legs; ML, male legs; FW, female wings; MW, male wings; FPr, female proboscis; MPr, male proboscis; Pg, pheromone gland. *P < 0.05.
Figure 9. Responses of Xenopus oocytes co-expressing EgriOR31/EgriORco and EgriOR1/EgriORco to stimulations with pheromone compounds. Inward current responses of EgriOR31/EgriORco and EgriOR1/EgriORco Xenopus oocytes in response to 10â5 M solutions of sex pheromone compounds.
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