Phylogenetic analysis of putative OR gene sequences of <i>O</i>. <i>emarginata</i> (black circle).

<p>The tree was rooted with Orco lineage (pink color). Bootstrap values < 50% are not shown. Color legend: Orange = PR group, yellow = OR18 group, green = OemaORs group, and blue = other general OR groups. Adis, <i>Athetis dissimilis</i>, Aips, <i>A</i>. <i>ipsilon</i>, Bmor, <i>B</i>. <i>mori</i>, Hvir, <i>H</i>. <i>virescens</i>, Msex, <i>M</i>. <i>sexta</i>, Oema, <i>O</i>. <i>emarginata</i>, Ofur, <i>O</i>. <i>furnacalis</i>, Slitu, <i>S</i>. <i>litura</i>.</p

Expression levels of olfactory genes in male and female antennae as measured by RT-qPCR analysis.

<p>Gene expression was calculated relative to the reference genes, <i>UCCR</i> and <i>AK</i>. The expression in female antennae was arbitrarily defined as 1 for all genes and was used in the normalization of gene expression of the male antennae. A, Expression levels of <i>CSP</i>, <i>IR</i>, and <i>SNMP</i> genes. B, Expression levels of the <i>OBP</i> genes. C, Expression levels of <i>OR</i> genes.</p

BLASTp results of candidate ionotropic receptors of <i>O</i>. <i>emarginata</i>.

<p>BLASTp results of candidate ionotropic receptors of <i>O</i>. <i>emarginata</i>.</p

Antennal transcriptome analysis of the piercing moth <i>Oraesia emarginata</i> (Lepidoptera: Noctuidae)

<div><p>The piercing fruit moth <i>Oraesia emarginata</i> is an economically significant pest; however, our understanding of its olfactory mechanisms in infestation is limited. The present study conducted antennal transcriptome analysis of olfactory genes using real-time quantitative reverse transcription PCR analysis (RT-qPCR). We identified a total of 104 candidate chemosensory genes from several gene families, including 35 olfactory receptors (ORs), 41 odorant-binding proteins, 20 chemosensory proteins, 6 ionotropic receptors, and 2 sensory neuron membrane proteins. Seven candidate pheromone receptors (PRs) and 3 candidate pheromone-binding proteins (PBPs) for sex pheromone recognition were found. <i>OemaOR29</i> and <i>OemaPBP1</i> had the highest fragments per kb per million fragments (FPKM) values in all ORs and OBPs, respectively. Eighteen olfactory genes were upregulated in females, including 5 candidate PRs, and 20 olfactory genes were upregulated in males, including 2 candidate PRs (<i>OemaOR29</i> and <i>4</i>) and 2 PBPs (<i>OemaPBP1</i> and <i>3</i>). These genes may have roles in mediating sex-specific behaviors. Most candidate olfactory genes of sex pheromone recognition (except <i>OemaOR29</i> and <i>OemaPBP3</i>) in <i>O</i>. <i>emarginata</i> were not clustered with those of studied noctuid species (type I pheromone). In addition, <i>OemaOR29</i> was belonged to cluster PRIII, which comprise proteins that recognize type II pheromones instead of type I pheromones. The structure and function of olfactory genes that encode sex pheromones in <i>O</i>. <i>emarginata</i> might thus differ from those of other studied noctuids. The findings of the present study may help explain the molecular mechanism underlying olfaction and the evolution of olfactory genes encoding sex pheromones in <i>O</i>. <i>emarginata</i>.</p></div

BLASTp results of candidate SNMP genes of <i>O</i>. <i>emarginata</i>.

<p>BLASTp results of candidate SNMP genes of <i>O</i>. <i>emarginata</i>.</p

BLASTp results of candidate olfactory receptors of <i>O</i>. <i>emarginata</i>.

<p>BLASTp results of candidate olfactory receptors of <i>O</i>. <i>emarginata</i>.</p

BLASTp results of candidate odorant-binding proteins of <i>O</i>. <i>emarginata</i>.

<p>BLASTp results of candidate odorant-binding proteins of <i>O</i>. <i>emarginata</i>.</p

Aligned putative full ORF of CSP gene sequences of <i>O</i>. <i>emarginata</i>.

<p>Four conserved cysteines are highlighted in blue.</p

Phylogenetic analysis of putative OBP gene sequences of <i>O</i>. <i>emarginata</i> (black circle), other moth species (black lines), and Dipteran species (green lines).

<p>The tree was rooted with the Lepidopteran GOBP-PBP group (green color). Bootstrap values < 50% are not shown. Color legend: Orange = conserved OBP groups, pink = expanded OemaOBPs group, green = Lepidopteran GOBP-PBP group, and blue = other general OBP groups. Adis, <i>A</i>. <i>dissimilis</i>, Agam, <i>Anopheles gambiae</i>, Aips, <i>A</i>. <i>ipsilon</i>, Bmor, <i>B</i>. <i>mori</i>, Cpun, <i>Conogethes punctiferalis</i>, Dmel, <i>Drosophila melanogaster</i>, Dple, <i>D</i>. <i>plexippus</i>, Gmol, <i>Grapholita molesta</i>, Harm, <i>H</i>. <i>armigera</i>, Hvir, <i>H</i>. <i>virescens</i>, Msex, <i>M</i>. <i>sexta</i>, Ofur, <i>O</i>. <i>furnacalis</i>, Oema, <i>O</i>. <i>emarginata</i>, Sexi, <i>S</i>. <i>exigua</i>, Slit, <i>S</i>. <i>littoralis</i>, Slitu, <i>S</i>. <i>litura</i>.</p

The phylogeny of Lepidopteran PBPs.

<p>The tree was rooted with GOBP lineage. Bootstrap values < 50% are not shown. Genes of <i>O</i>. <i>emarginata</i>, other species with type II pheromones, and the other noctuid species are indicated by black circles, black triangles, and diamonds, respectively. Clusters PBPI—PBPIII are indicated by orange, purple, and blue colors, respectively. Acon, <i>Argyresthia conjugella</i>, Aips, <i>A</i>. <i>ipsilon</i>, Apol, <i>A</i>. <i>polyphemus</i>, Asel, <i>Ascotis selenaria cretacea</i>, Bmor, <i>B</i>. <i>mori</i>, Cpun, <i>C</i>. <i>punctiferalis</i>, Csup, <i>C</i>. <i>suppressalis</i>, Ehip, <i>Eogystia hippophaecolus</i>, Harm, <i>H</i>. <i>armigera</i>, Hass, <i>H</i>. <i>assulta</i>, Gmol, <i>G</i>. <i>molesta</i>, Ldis, <i>Lymantria dispar</i>, Msex, <i>M</i>. <i>sexta</i>, Obru, <i>O</i>. <i>brumata</i>, Oema, <i>O</i>. <i>emarginata</i>, Ofur, <i>O</i>. <i>furnacalis</i>, Onub, <i>O</i>. <i>nubilalis</i>, Pxyl, <i>P</i>. <i>xylostella</i>, Sexi, <i>S</i>. <i>exigua</i>, Sinf, <i>S</i>. <i>inferens</i>, Slit, <i>S</i>. <i>litura</i>.</p