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Identification and comparative expression analysis of odorant binding protein genes in the tobacco cutworm Spodoptera litura.

Gu SH, Zhou JJ, Gao S, Wang DH, Li XC, Guo YY, Zhang YJ - Sci Rep (2015)

Bottom Line: Some regular patterns and key conserved motifs of OBPs in genus Spodoptera are identified by MEME, and their putative roles in detecting odorants are discussed here.The motif-patterns between Lepidoptera OBPs and CSPs are also compared.The SlitOBPs identified here provide a starting point to facilitate functional studies of insect OBPs at the molecular level both in vivo and in vitro.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, China.

ABSTRACT
Insect odorant binding proteins (OBPs) are thought to involve in insects' olfaction perception. In the present study, we identified 38 OBP genes from the antennal transcriptomes of Spodoptera litura. Tissue expression profiles analysis revealed that 17 of the 38 SlitOBP transcripts were uniquely or primarily expressed in the antennae of both sexes, suggesting their putative role in chemoreception. The RPKM value analysis revealed that seven OBPs (SlitPBP1-3, SlitGOBP1-2, SlitOBP3 and SlitOBP5) are highly abundant in male and female antennae. Most S. litura antennal unigenes had high homology with Lepidoptera insects, especially genes of the genus Spodoptera. Phylogenetic analysis of the Lepidoptera OBPs demonstrated that the OBP genes from the genus Spodoptera (S. litura, Spodoptera littoralis and Spodoptera exigua) had a relatively close evolutionary relationship. Some regular patterns and key conserved motifs of OBPs in genus Spodoptera are identified by MEME, and their putative roles in detecting odorants are discussed here. The motif-patterns between Lepidoptera OBPs and CSPs are also compared. The SlitOBPs identified here provide a starting point to facilitate functional studies of insect OBPs at the molecular level both in vivo and in vitro.

No MeSH data available.


S. litura OBP transcript levels in different tissues as measured by RT-qPCR.MA: male antennae; FA: female antennae. The internal controls β-actin and ribosomal protein L31 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 L31 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).
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f9: S. litura OBP transcript levels in different tissues as measured by RT-qPCR.MA: male antennae; FA: female antennae. The internal controls β-actin and ribosomal protein L31 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 L31 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).

Mentions: In order to confirm the RT-PCR results, real-time quantitative PCR (RT-qPCR) analyses were performed to measure quantitatively expression levels of the 38 OBP genes in the male antennae, female antennae and body parts (mixture of heads, thoraxes, abdomens, legs, wings) (Fig. 9). The results confirmed the transcript expression level of 17 of the 38 SlitOBP genes (SlitPBP1-3, SlitGOBP1-2, SlitOBP1-5, SlitOBP7-9, SlitOBP14-15, SlitOBP17 and SlitOBP21) was approximately 30 to 50000 times higher in both the male and female antennae than in the body parts. Furthermore, the three antennae-specific PBPs (SlitPBP1-3) showed an expression level of 6.8, 7.9 and 6.4 times higher in the male antennae than in the female antennae, respectively (p < 0.01), and the expression level of 7 antennae-specific OBPs (SlitGOBP1-2, SlitOBP2, SlitOBP8, SlitOBP9, SlitOBP17, SlitOBP21) was 4.0, 3.4, 2.8, 2.4, 1.6, 1.6 and 2.3 times higher in the female antennae than in the male antennae (p < 0.01). Seven OBPs (SlitOBP1, SlitOBP3-7, SlitOBP15) were found to be mainly expressed in the antennae and showed a similar expression level between the sexes (p > 0.01). Three OBPs (SlitOBP19, SlitOBP20 and SlitOBP33) were detected mainly expressed in the body, with the expression levels 23 to 200 times higher than in the antennae. The remaining 18 OBPs (SlitOBP6, SlitOBP10-13, SlitOBP16, SlitOBP18, SlitOBP22-32) showed similar expression levels in the male antennae, female antennae and body parts (p > 0.01).


Identification and comparative expression analysis of odorant binding protein genes in the tobacco cutworm Spodoptera litura.

Gu SH, Zhou JJ, Gao S, Wang DH, Li XC, Guo YY, Zhang YJ - Sci Rep (2015)

S. litura OBP transcript levels in different tissues as measured by RT-qPCR.MA: male antennae; FA: female antennae. The internal controls β-actin and ribosomal protein L31 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 L31 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).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4561897&req=5

f9: S. litura OBP transcript levels in different tissues as measured by RT-qPCR.MA: male antennae; FA: female antennae. The internal controls β-actin and ribosomal protein L31 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 L31 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).
Mentions: In order to confirm the RT-PCR results, real-time quantitative PCR (RT-qPCR) analyses were performed to measure quantitatively expression levels of the 38 OBP genes in the male antennae, female antennae and body parts (mixture of heads, thoraxes, abdomens, legs, wings) (Fig. 9). The results confirmed the transcript expression level of 17 of the 38 SlitOBP genes (SlitPBP1-3, SlitGOBP1-2, SlitOBP1-5, SlitOBP7-9, SlitOBP14-15, SlitOBP17 and SlitOBP21) was approximately 30 to 50000 times higher in both the male and female antennae than in the body parts. Furthermore, the three antennae-specific PBPs (SlitPBP1-3) showed an expression level of 6.8, 7.9 and 6.4 times higher in the male antennae than in the female antennae, respectively (p < 0.01), and the expression level of 7 antennae-specific OBPs (SlitGOBP1-2, SlitOBP2, SlitOBP8, SlitOBP9, SlitOBP17, SlitOBP21) was 4.0, 3.4, 2.8, 2.4, 1.6, 1.6 and 2.3 times higher in the female antennae than in the male antennae (p < 0.01). Seven OBPs (SlitOBP1, SlitOBP3-7, SlitOBP15) were found to be mainly expressed in the antennae and showed a similar expression level between the sexes (p > 0.01). Three OBPs (SlitOBP19, SlitOBP20 and SlitOBP33) were detected mainly expressed in the body, with the expression levels 23 to 200 times higher than in the antennae. The remaining 18 OBPs (SlitOBP6, SlitOBP10-13, SlitOBP16, SlitOBP18, SlitOBP22-32) showed similar expression levels in the male antennae, female antennae and body parts (p > 0.01).

Bottom Line: Some regular patterns and key conserved motifs of OBPs in genus Spodoptera are identified by MEME, and their putative roles in detecting odorants are discussed here.The motif-patterns between Lepidoptera OBPs and CSPs are also compared.The SlitOBPs identified here provide a starting point to facilitate functional studies of insect OBPs at the molecular level both in vivo and in vitro.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, China.

ABSTRACT
Insect odorant binding proteins (OBPs) are thought to involve in insects' olfaction perception. In the present study, we identified 38 OBP genes from the antennal transcriptomes of Spodoptera litura. Tissue expression profiles analysis revealed that 17 of the 38 SlitOBP transcripts were uniquely or primarily expressed in the antennae of both sexes, suggesting their putative role in chemoreception. The RPKM value analysis revealed that seven OBPs (SlitPBP1-3, SlitGOBP1-2, SlitOBP3 and SlitOBP5) are highly abundant in male and female antennae. Most S. litura antennal unigenes had high homology with Lepidoptera insects, especially genes of the genus Spodoptera. Phylogenetic analysis of the Lepidoptera OBPs demonstrated that the OBP genes from the genus Spodoptera (S. litura, Spodoptera littoralis and Spodoptera exigua) had a relatively close evolutionary relationship. Some regular patterns and key conserved motifs of OBPs in genus Spodoptera are identified by MEME, and their putative roles in detecting odorants are discussed here. The motif-patterns between Lepidoptera OBPs and CSPs are also compared. The SlitOBPs identified here provide a starting point to facilitate functional studies of insect OBPs at the molecular level both in vivo and in vitro.

No MeSH data available.