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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

Neighbor-joining tree of candidate odorant receptor (OR) and gustatory receptor (GR) from G. molesta and other Lepidoptera.The tree was drawn using Adobe Photoshop CS5, based on an unrooted tree constructed using the BioNJ algorithm in Seaview v.4. The unrooted tree was constructed based on the sequence alignment produced using MAFFT version 6. Gmol, Grapholita molesta, Msex, Manduca sexta, Cpom, Cydia pomonella, Slit, Spodoptera Iittoralis, Bmor, Bombyx mori.
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pone.0142193.g008: Neighbor-joining tree of candidate odorant receptor (OR) and gustatory receptor (GR) from G. molesta and other Lepidoptera.The tree was drawn using Adobe Photoshop CS5, based on an unrooted tree constructed using the BioNJ algorithm in Seaview v.4. The unrooted tree was constructed based on the sequence alignment produced using MAFFT version 6. Gmol, Grapholita molesta, Msex, Manduca sexta, Cpom, Cydia pomonella, Slit, Spodoptera Iittoralis, Bmor, Bombyx mori.

Mentions: Bioinformatic analysis identified 48 different sequences encoding putative ORs and four sequences encoding putative GRs. GmolORs and GmolGRs were named according to their similarities with previously annotated Lepidoptera ORs and the topology predictions from TMpred as observed from other insect ORs [45]. Twelve of these sequences appeared to contain full lengths genes since they had full length ORFs with 5–8 transmembrane domains (Table 5). The co-receptor of G.molesta showed 95% identity to C. pomonella co-receptor, CpomOR2, one of the most conserved co-receptors in insect species. While similar with other insect ORs, most GmolORs were highly divergent and shared low similarity with other insect ORs, except for closely related species such as Cydia pomonella. Four candidate GmolORs sequences (GmolOR1, GmolOR4, GmolOR6, and GmolOR11) tended to be pheromone receptors (PRs) as they are highly conserved with CpomOR fragments and BmorPRs amino acid sequences from other species [46]. These four sequences were clusterd into one subgroup in the phylogenetic tree (Fig 8). The gustatory receptors that we identified (GmolGR2, GmolGR3, and GmolGR4) were found in a clade with sugar receptors, which included gustatory receptors identified from other moth antennae; these gustatory receptors were also clustered in this clade (Fig 8) [39,47,48]. Another putative GR, named GmolGR1, was clustered with putative CO2 receptors. The information of ORs and GRs are given in Table 5, 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)

Neighbor-joining tree of candidate odorant receptor (OR) and gustatory receptor (GR) from G. molesta and other Lepidoptera.The tree was drawn using Adobe Photoshop CS5, based on an unrooted tree constructed using the BioNJ algorithm in Seaview v.4. The unrooted tree was constructed based on the sequence alignment produced using MAFFT version 6. Gmol, Grapholita molesta, Msex, Manduca sexta, Cpom, Cydia pomonella, Slit, Spodoptera Iittoralis, Bmor, Bombyx mori.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0142193.g008: Neighbor-joining tree of candidate odorant receptor (OR) and gustatory receptor (GR) from G. molesta and other Lepidoptera.The tree was drawn using Adobe Photoshop CS5, based on an unrooted tree constructed using the BioNJ algorithm in Seaview v.4. The unrooted tree was constructed based on the sequence alignment produced using MAFFT version 6. Gmol, Grapholita molesta, Msex, Manduca sexta, Cpom, Cydia pomonella, Slit, Spodoptera Iittoralis, Bmor, Bombyx mori.
Mentions: Bioinformatic analysis identified 48 different sequences encoding putative ORs and four sequences encoding putative GRs. GmolORs and GmolGRs were named according to their similarities with previously annotated Lepidoptera ORs and the topology predictions from TMpred as observed from other insect ORs [45]. Twelve of these sequences appeared to contain full lengths genes since they had full length ORFs with 5–8 transmembrane domains (Table 5). The co-receptor of G.molesta showed 95% identity to C. pomonella co-receptor, CpomOR2, one of the most conserved co-receptors in insect species. While similar with other insect ORs, most GmolORs were highly divergent and shared low similarity with other insect ORs, except for closely related species such as Cydia pomonella. Four candidate GmolORs sequences (GmolOR1, GmolOR4, GmolOR6, and GmolOR11) tended to be pheromone receptors (PRs) as they are highly conserved with CpomOR fragments and BmorPRs amino acid sequences from other species [46]. These four sequences were clusterd into one subgroup in the phylogenetic tree (Fig 8). The gustatory receptors that we identified (GmolGR2, GmolGR3, and GmolGR4) were found in a clade with sugar receptors, which included gustatory receptors identified from other moth antennae; these gustatory receptors were also clustered in this clade (Fig 8) [39,47,48]. Another putative GR, named GmolGR1, was clustered with putative CO2 receptors. The information of ORs and GRs are given in Table 5, 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