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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 evaluated by RT-PCR.MA: male antennae; FA: female antennae; H: heads; T: thoraxes; A: abdomens; L: legs; W: wings. 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. The RT-PCR for all SlitOBPs are run under the same experimental conditions. The display for each SlitOBPs are cropped figures from the gels. The full-length gels are presented in Supplementary Figure 1.
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f8: S. litura OBP transcript levels in different tissues as evaluated by RT-PCR.MA: male antennae; FA: female antennae; H: heads; T: thoraxes; A: abdomens; L: legs; W: wings. 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. The RT-PCR for all SlitOBPs are run under the same experimental conditions. The display for each SlitOBPs are cropped figures from the gels. The full-length gels are presented in Supplementary Figure 1.

Mentions: The expression of the 38 identified SlitOBP genes in different tissue types were examined using RT-PCR (Fig. 8). The results indicated that 17 of the 38 SlitOBP genes (SlitPBP1-3, SlitGOBP1-2, SlitOBP1-5, SlitOBP7-9, SlitOBP14-15, SlitOBP17 and SlitOBP21) were uniquely or primarily expressed in the male and female antennae; three OBPs SlitOBP19, SlitOBP20 and SlitOBP33 were only detected in the abdomen; the remaining 18 OBPs (SlitOBP6, SlitOBP10-13, SlitOBP16, SlitOBP18, SlitOBP22-32) were expressed not only in the antennae but also in other tissues such as heads, thoraxes, abdomens, legs and wings (Fig. 8). Equal amount cDNA (200 ng) were used in the RT-PCR reactions, the intensity of the PCR bands in antennae of some SlitOBPs was very weak or undetectable, such as SlitOBP9, SlitOBP10, SlitOBP19-20, SlitOBP22-33, the reason for this may be the relatively low expression levels of these SlitOBPs in the antennae, which consistent with the results of the low abundance of these SlitOBPs (RKKM value < 20) (Table 2).


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 evaluated by RT-PCR.MA: male antennae; FA: female antennae; H: heads; T: thoraxes; A: abdomens; L: legs; W: wings. 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. The RT-PCR for all SlitOBPs are run under the same experimental conditions. The display for each SlitOBPs are cropped figures from the gels. The full-length gels are presented in Supplementary Figure 1.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f8: S. litura OBP transcript levels in different tissues as evaluated by RT-PCR.MA: male antennae; FA: female antennae; H: heads; T: thoraxes; A: abdomens; L: legs; W: wings. 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. The RT-PCR for all SlitOBPs are run under the same experimental conditions. The display for each SlitOBPs are cropped figures from the gels. The full-length gels are presented in Supplementary Figure 1.
Mentions: The expression of the 38 identified SlitOBP genes in different tissue types were examined using RT-PCR (Fig. 8). The results indicated that 17 of the 38 SlitOBP genes (SlitPBP1-3, SlitGOBP1-2, SlitOBP1-5, SlitOBP7-9, SlitOBP14-15, SlitOBP17 and SlitOBP21) were uniquely or primarily expressed in the male and female antennae; three OBPs SlitOBP19, SlitOBP20 and SlitOBP33 were only detected in the abdomen; the remaining 18 OBPs (SlitOBP6, SlitOBP10-13, SlitOBP16, SlitOBP18, SlitOBP22-32) were expressed not only in the antennae but also in other tissues such as heads, thoraxes, abdomens, legs and wings (Fig. 8). Equal amount cDNA (200 ng) were used in the RT-PCR reactions, the intensity of the PCR bands in antennae of some SlitOBPs was very weak or undetectable, such as SlitOBP9, SlitOBP10, SlitOBP19-20, SlitOBP22-33, the reason for this may be the relatively low expression levels of these SlitOBPs in the antennae, which consistent with the results of the low abundance of these SlitOBPs (RKKM value < 20) (Table 2).

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.