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. 2014 Aug 1;9(8):e103420.
doi: 10.1371/journal.pone.0103420. eCollection 2014.

Molecular characterization and differential expression of olfactory genes in the antennae of the black cutworm moth Agrotis ipsilon

Affiliations

Molecular characterization and differential expression of olfactory genes in the antennae of the black cutworm moth Agrotis ipsilon

Shao-Hua Gu et al. PLoS One. .

Abstract

Insects use their sensitive and selective olfactory system to detect outside chemical odorants, such as female sex pheromones and host plant volatiles. Several groups of olfactory proteins participate in the odorant detection process, including odorant binding proteins (OBPs), chemosensory proteins (CSPs), odorant receptors (ORs), ionotropic receptors (IRs) and sensory neuron membrane proteins (SNMPs). The identification and functional characterization of these olfactory proteins will enhance our knowledge of the molecular basis of insect chemoreception. In this study, we report the identification and differential expression profiles of these olfactory genes in the black cutworm moth Agrotis ipsilon. In total, 33 OBPs, 12 CSPs, 42 ORs, 24 IRs, 2 SNMPs and 1 gustatory receptor (GR) were annotated from the A. ipsilon antennal transcriptomes, and further RT-PCR and RT-qPCR revealed that 22 OBPs, 3 CSPs, 35 ORs, 14 IRs and the 2 SNMPs are uniquely or primarily expressed in the male and female antennae. Furthermore, one OBP (AipsOBP6) and one CSP (AipsCSP2) were exclusively expressed in the female sex pheromone gland. These antennae-enriched OBPs, CSPs, ORs, IRs and SNMPs were suggested to be responsible for pheromone and general odorant detection and thus could be meaningful target genes for us to study their biological functions in vivo and in vitro.

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Conflict of interest statement

Competing Interests: The authors declare that they have no competing interests.

Figures

Figure 1
Figure 1. The size distribution of the clean reads and assembled unigenes from A. ipsilon male and female antennal transcriptomes.
Figure 2
Figure 2. Top 20 best hits of the BLASTn results.
All A. ipsilon antennal unigenes were used in BLASTn to search the GenBank entries. The best hits with an E-value < = 1.0E-5 for each query were grouped according to species.
Figure 3
Figure 3. Gene Ontology (GO) classifications of the male and female A. ipsilon antennal unigenes according to their involvement in biological processes, cellular component and molecular function.
Figure 4
Figure 4. A. ipsilon OBP and CSP transcript levels in different tissues as evaluated by RT-PCR.
MA: male antennae; FA: female antennae; Bo: body. Pheromone gland rather than body was used in the analysis of AipsOBP6 and AipsCSP2. Antennae specific or enriched genes are labeled with a red pentagram. β-actin was used as an internal reference gene to test the integrity of each cDNA template; the similar intensity of β-actin bands among different tissues indicates the use of equal template concentrations.
Figure 5
Figure 5. A. ipsilon OBP transcript levels in different tissues as measured by RT-qPCR.
MA: male antennae; FA: female antennae; Bo: body. Pheromone gland rather than body was used in the analysis of AipsOBP6. The internal controls β-actin and ribosomal protein S3 were used to normalize transcript levels in each sample. This figure was presented using β-actin as the reference gene to normalize the target gene expression and to correct sample-to-sample variation; similar results were obtained with ribosomal protein S3 as the reference gene. The standard error is represented by the error bar, and the different letters (a, b, c) above each bar denote significant differences (p<0.05).
Figure 6
Figure 6. A. ipsilon CSP transcript levels in different tissues as measured by RT-qPCR.
MA: male antennae; FA: female antennae; Bo: body. Pheromone gland rather than body was used in the analysis of AipsCSP2. The internal controls β-actin and ribosomal protein S3 were used to normalize transcript levels in each sample. This figure was presented using β-actin as reference gene to normalize the target gene expression and correct sample-to-sample variation; similar results were obtained with ribosomal protein S3 as the reference gene. The standard error is represented by the error bar, and the different letters (a, b) above each bar denote significant differences (p<0.05).
Figure 7
Figure 7. A. ipsilon OR, IR, SNMP and GR transcript levels in different tissues as evaluated by RT-PCR.
MA: male antennae; FA: female antennae; Bo: body. Genes that are equally expressed in the male and female antennae are labeled with a red pentagram. Genes that are specifically or primarily expressed in the male antennae are labeled with a red circle. Genes that are specifically or primarily expressed in the female antennae are labeled with a red triangle. β-actin was used as the internal reference gene to test the integrity of each cDNA templates; the similar intensity of β-actin bands among different tissues indicates the use of equal template concentrations.
Figure 8
Figure 8. A. ipsilon OR transcript levels in different tissues as measured by RT-qPCR.
MA: male antennae; FA: female antennae; Bo: body. The internal controls β-actin and ribosomal protein S3 were used to normalize transcript levels in each sample. This figure was presented using β-actin as the reference gene to normalize the target gene expression and to correct sample-to-sample variation; similar results were obtained with ribosomal protein S3 as the reference gene. The standard error is represented by the error bar, and the different letters (a, b, c) above each bar denote significant differences (p<0.05).
Figure 9
Figure 9. Neighbor-joining tree of candidate odorant receptor proteins from A. ipsilon (red), B. mori (green) and H. virescens (blue).
The protein names and sequences of ORs that were used in this analysis are listed in Table S5.
Figure 10
Figure 10. A. ipsilon IR, SNMP and GR transcript levels in different tissues as measured by RT-qPCR.
MA: male antennae; FA: female antennae; Bo: body. The internal controls β-actin and ribosomal protein S3 were used to normalize transcript levels in each sample. This figure was presented using β-actin as the reference gene to normalize the target gene expression and to correct sample-to-sample variation; similar results were obtained with ribosomal protein S3 as the reference gene. The standard error is represented by the error bar, and the different letters (a, b, c) above each bar denote significant differences (p<0.05).
Figure 11
Figure 11. Neighbor-joining tree of candidate ionotropic receptor proteins from different insect species.
The protein names and sequences of IRs that were used in this analysis are listed in Table S6.

References

    1. Kaissling KE (1986) Chemo-electrical transduction in insect olfaction receptors. Annu Rev Neurosci 9: 121–145. - PubMed
    1. Steinbrecht RA (1997) Pore structures in insect olfactory sensilla: a review of data and concepts. Int J Insect Morphol Embryol 26: 229–245.
    1. Leal WS (2013) Odorant reception in insects: roles of receptors, binding proteins, and degrading enzymes. Annu Rev Entomol 58: 373–391. - PubMed
    1. Vogt RG, Riddiford LM (1981) Pheromone binding and inactivation by moth antennae. Nature 293: 161–163. - PubMed
    1. Klein U (1987) Sensillum-lymph proteins from antennal olfactory hairs of the moth Antheraea polyphemus (Saturniidae). Insect Biochem 17: 1193–1204.

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