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Identification of Putative Olfactory Genes from the Oriental Fruit Moth Grapholita molesta via an Antennal Transcriptome Analysis.

Li G, Du J, Li Y, Wu J - PLoS ONE (2015)

Bottom Line: The identified genes were further confirmed through semi-quantitative reverse transcription PCR for transcripts coding for 26 OBPs and 17 CSPs.OBP transcripts showed an obvious antenna bias, whereas CSP transcripts were detected in different tissues.Antennal transcriptome data derived from the oriental fruit moth constituted an abundant molecular resource for the identification of genes potentially involved in the olfaction process of the species.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Plant Protection Resources and Pest Management (Northwest A&F University), Ministry of Education, Yangling, Shaanxi, People's Republic of China.

ABSTRACT

Background: The oriental fruit moth, Grapholita molesta, is an extremely important oligophagous pest species of stone and pome fruits throughout the world. As a host-switching species, adult moths, especially females, depend on olfactory cues to a large extent in locating host plants, finding mates, and selecting oviposition sites. The identification of olfactory genes can facilitate investigation on mechanisms for chemical communications.

Methodology/principal finding: We generated transcriptome of female antennae of G.molesta using the next-generation sequencing technique, and assembled transcripts from RNA-seq reads using Trinity, SOAPdenovo-trans and Abyss-trans assemblers. We identified 124 putative olfactory genes. Among the identified olfactory genes, 118 were novel to this species, including 28 transcripts encoding for odorant binding proteins, 17 chemosensory proteins, 48 odorant receptors, four gustatory receptors, 24 ionotropic receptors, two sensory neuron membrane proteins, and one odor degrading enzyme. The identified genes were further confirmed through semi-quantitative reverse transcription PCR for transcripts coding for 26 OBPs and 17 CSPs. OBP transcripts showed an obvious antenna bias, whereas CSP transcripts were detected in different tissues.

Conclusion: Antennal transcriptome data derived from the oriental fruit moth constituted an abundant molecular resource for the identification of genes potentially involved in the olfaction process of the species. This study provides a foundation for future research on the molecules involved in olfactory recognition of this insect pest, and in particular, the feasibility of using semiochemicals to control this pest.

No MeSH data available.


Related in: MedlinePlus

Sequences alignment of candidate GmolOBPs.The six conserved cysteine residues were marked with “☆”. As GmolOBP17, 18, 19, 20, 21, 22 and 23 are not intact sequences, those sequences are not included in the multisequence alignment.
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pone.0142193.g004: Sequences alignment of candidate GmolOBPs.The six conserved cysteine residues were marked with “☆”. As GmolOBP17, 18, 19, 20, 21, 22 and 23 are not intact sequences, those sequences are not included in the multisequence alignment.

Mentions: First, we used motif scanning to detect the conserved six cysteine residues pattern (C1-X20-66-C2-X3-C3-X21-43-C4-X8-14-C5-X8-C6 or C1-X15-39-C2-X3-C3-X21-44-C4-X7-12-C5-X8-C6, where X is any amino acid) of the candidate odorant-binding proteins [42]. We then used keyword searching and PSI-Blast. Twenty-eight sequences encoding putative odorant-binding proteins were identified, including two GOBPs and three PBPs. Of the 28 sequences, 21 had full ORFs, four unigenes had full-length ORFs, but without a signal peptide. Sequence alignment showed that almost all the putative OBPs shared the classic six-cysteine motif, except GmolOBP14, which was grouped into the “minus-C” subgroup with the second and fifth cysteine residues missing [43] (Fig 4). In the phylogenetic tree, as expected, the PBP and GOBP sequences were clustered into separate clades away from other OBPs. All the candidate OBP sequences with at least one lepidopteran ortholog were clustered in congruence with the BLAST results (Fig 5). Comparing our candidate OBPs with previously recorded OBPs of G.molesta in NCBI, 23 sequences as new genes, including GmolPBP1, GmolOBP1, GmolOBP2, and GmolOBP4 to GmolOBP23. The information on the OBPs is listed in Table 3. The nucleotide sequences are listed in S2 File.


Identification of Putative Olfactory Genes from the Oriental Fruit Moth Grapholita molesta via an Antennal Transcriptome Analysis.

Li G, Du J, Li Y, Wu J - PLoS ONE (2015)

Sequences alignment of candidate GmolOBPs.The six conserved cysteine residues were marked with “☆”. As GmolOBP17, 18, 19, 20, 21, 22 and 23 are not intact sequences, those sequences are not included in the multisequence alignment.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4635014&req=5

pone.0142193.g004: Sequences alignment of candidate GmolOBPs.The six conserved cysteine residues were marked with “☆”. As GmolOBP17, 18, 19, 20, 21, 22 and 23 are not intact sequences, those sequences are not included in the multisequence alignment.
Mentions: First, we used motif scanning to detect the conserved six cysteine residues pattern (C1-X20-66-C2-X3-C3-X21-43-C4-X8-14-C5-X8-C6 or C1-X15-39-C2-X3-C3-X21-44-C4-X7-12-C5-X8-C6, where X is any amino acid) of the candidate odorant-binding proteins [42]. We then used keyword searching and PSI-Blast. Twenty-eight sequences encoding putative odorant-binding proteins were identified, including two GOBPs and three PBPs. Of the 28 sequences, 21 had full ORFs, four unigenes had full-length ORFs, but without a signal peptide. Sequence alignment showed that almost all the putative OBPs shared the classic six-cysteine motif, except GmolOBP14, which was grouped into the “minus-C” subgroup with the second and fifth cysteine residues missing [43] (Fig 4). In the phylogenetic tree, as expected, the PBP and GOBP sequences were clustered into separate clades away from other OBPs. All the candidate OBP sequences with at least one lepidopteran ortholog were clustered in congruence with the BLAST results (Fig 5). Comparing our candidate OBPs with previously recorded OBPs of G.molesta in NCBI, 23 sequences as new genes, including GmolPBP1, GmolOBP1, GmolOBP2, and GmolOBP4 to GmolOBP23. The information on the OBPs is listed in Table 3. The nucleotide sequences are listed in S2 File.

Bottom Line: The identified genes were further confirmed through semi-quantitative reverse transcription PCR for transcripts coding for 26 OBPs and 17 CSPs.OBP transcripts showed an obvious antenna bias, whereas CSP transcripts were detected in different tissues.Antennal transcriptome data derived from the oriental fruit moth constituted an abundant molecular resource for the identification of genes potentially involved in the olfaction process of the species.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Plant Protection Resources and Pest Management (Northwest A&F University), Ministry of Education, Yangling, Shaanxi, People's Republic of China.

ABSTRACT

Background: The oriental fruit moth, Grapholita molesta, is an extremely important oligophagous pest species of stone and pome fruits throughout the world. As a host-switching species, adult moths, especially females, depend on olfactory cues to a large extent in locating host plants, finding mates, and selecting oviposition sites. The identification of olfactory genes can facilitate investigation on mechanisms for chemical communications.

Methodology/principal finding: We generated transcriptome of female antennae of G.molesta using the next-generation sequencing technique, and assembled transcripts from RNA-seq reads using Trinity, SOAPdenovo-trans and Abyss-trans assemblers. We identified 124 putative olfactory genes. Among the identified olfactory genes, 118 were novel to this species, including 28 transcripts encoding for odorant binding proteins, 17 chemosensory proteins, 48 odorant receptors, four gustatory receptors, 24 ionotropic receptors, two sensory neuron membrane proteins, and one odor degrading enzyme. The identified genes were further confirmed through semi-quantitative reverse transcription PCR for transcripts coding for 26 OBPs and 17 CSPs. OBP transcripts showed an obvious antenna bias, whereas CSP transcripts were detected in different tissues.

Conclusion: Antennal transcriptome data derived from the oriental fruit moth constituted an abundant molecular resource for the identification of genes potentially involved in the olfaction process of the species. This study provides a foundation for future research on the molecules involved in olfactory recognition of this insect pest, and in particular, the feasibility of using semiochemicals to control this pest.

No MeSH data available.


Related in: MedlinePlus