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Chemosensillum immunolocalization and ligand specificity of chemosensory proteins in the alfalfa plant bug Adelphocoris lineolatus (Goeze).

Sun L, Zhou JJ, Gu SH, Xiao HJ, Guo YY, Liu ZW, Zhang YJ - Sci Rep (2015)

Bottom Line: Further cellular investigation of their immunolocalization revealed their dynamic protein expression profiles among different types of antennal sensilla.In ad`dition, a cooperative interaction was observed between two co-expressed CSPs resulting in an increase of the binding affinities by a mixture of AlinCSP5 and AlinCSP6 to terpenoids which do not bind to individual CSPs.These findings in combination with our previous data for AlinCSP1-3 indicate a possible functional differentiation of CSPs in the A. lineolatus olfactory system.

View Article: PubMed Central - PubMed

Affiliation: 1] State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, 100193, China [2] Key Laboratory of Tea Plants Biology and Resources Utilization of Agriculture Ministry, Tea Research Institute, Chinese Academy of Agricultural Sciences, Hangzhou, 310008, China [3] Key Laboratory of Integrated Management of Crop Diseases and Pests (Ministry of Education), College of Plant Protection, Nanjing Agricultural University, Nanjing, 210095, China.

ABSTRACT
Insect chemosensory proteins (CSPs) are a family of small soluble proteins. To date, their physiological functions in insect olfaction remain largely controversial in comparison to odorant binding proteins (OBPs). In present study, we reported the antenna specific expression of three CSPs (AlinCSP4-6) from Adelphocoris lineolatus, their distinct chemosensillum distribution as well as ligand binding capability thus providing the evidence for the possible roles that they could play in semiochemical detection of the plant bug A. lineolatus. The results of qRT-PCR and western blot assay clearly showed that all of these three CSPs are highly expressed in the adult antennae, the olfactory organ of insects. Further cellular investigation of their immunolocalization revealed their dynamic protein expression profiles among different types of antennal sensilla. In a fluorescence competitive binding assay, the selective ligand binding was observed for AlinCSP4-6. In ad`dition, a cooperative interaction was observed between two co-expressed CSPs resulting in an increase of the binding affinities by a mixture of AlinCSP5 and AlinCSP6 to terpenoids which do not bind to individual CSPs. These findings in combination with our previous data for AlinCSP1-3 indicate a possible functional differentiation of CSPs in the A. lineolatus olfactory system.

No MeSH data available.


SDS-PAGE and protein expression profiles among different adult tissues of both sexes measured by western blot analysis.From left to right, SDS-PAGE of crude extracts of different adult tissues of both sexes (1st panel) and western blot analysis (2nd–4th panels).
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f3: SDS-PAGE and protein expression profiles among different adult tissues of both sexes measured by western blot analysis.From left to right, SDS-PAGE of crude extracts of different adult tissues of both sexes (1st panel) and western blot analysis (2nd–4th panels).

Mentions: To characterize the immunolocalization of AlinCSP4-6 proteins among different antennae sensilla of A. lineolatus, polyclonal antisera against each of recombinant AlinCSP4-6 proteins were produced. A western blot assay of the purified recombinant proteins suggested that each antiserum is specific and sufficient to distinguish from one another without cross reactions because anti-AlinCSP4, anti-AlinCSP5, and anti-AlinCSP6 antibody specifically reacted with recombinant AlinCSP4, AlinCSP5, and AlinCSP6, respectively (Figure S3). The tissue specific distributions of AlinCSP4-6 proteins were then analyzed by the western blot of crude extracts from different tissues. The results showed that both AlinCSP4 and AlinCSP5 were antennae-specific in male and female, whereas AlinCSP6 was highly expressed in the antennae and legs in both sexes and weakly expressed in the male wings (Figure 3). There was no detectable AlinCSP4-6 protein expression in the stylets and any of other non-olfactory tissues.


Chemosensillum immunolocalization and ligand specificity of chemosensory proteins in the alfalfa plant bug Adelphocoris lineolatus (Goeze).

Sun L, Zhou JJ, Gu SH, Xiao HJ, Guo YY, Liu ZW, Zhang YJ - Sci Rep (2015)

SDS-PAGE and protein expression profiles among different adult tissues of both sexes measured by western blot analysis.From left to right, SDS-PAGE of crude extracts of different adult tissues of both sexes (1st panel) and western blot analysis (2nd–4th panels).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4308698&req=5

f3: SDS-PAGE and protein expression profiles among different adult tissues of both sexes measured by western blot analysis.From left to right, SDS-PAGE of crude extracts of different adult tissues of both sexes (1st panel) and western blot analysis (2nd–4th panels).
Mentions: To characterize the immunolocalization of AlinCSP4-6 proteins among different antennae sensilla of A. lineolatus, polyclonal antisera against each of recombinant AlinCSP4-6 proteins were produced. A western blot assay of the purified recombinant proteins suggested that each antiserum is specific and sufficient to distinguish from one another without cross reactions because anti-AlinCSP4, anti-AlinCSP5, and anti-AlinCSP6 antibody specifically reacted with recombinant AlinCSP4, AlinCSP5, and AlinCSP6, respectively (Figure S3). The tissue specific distributions of AlinCSP4-6 proteins were then analyzed by the western blot of crude extracts from different tissues. The results showed that both AlinCSP4 and AlinCSP5 were antennae-specific in male and female, whereas AlinCSP6 was highly expressed in the antennae and legs in both sexes and weakly expressed in the male wings (Figure 3). There was no detectable AlinCSP4-6 protein expression in the stylets and any of other non-olfactory tissues.

Bottom Line: Further cellular investigation of their immunolocalization revealed their dynamic protein expression profiles among different types of antennal sensilla.In ad`dition, a cooperative interaction was observed between two co-expressed CSPs resulting in an increase of the binding affinities by a mixture of AlinCSP5 and AlinCSP6 to terpenoids which do not bind to individual CSPs.These findings in combination with our previous data for AlinCSP1-3 indicate a possible functional differentiation of CSPs in the A. lineolatus olfactory system.

View Article: PubMed Central - PubMed

Affiliation: 1] State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, 100193, China [2] Key Laboratory of Tea Plants Biology and Resources Utilization of Agriculture Ministry, Tea Research Institute, Chinese Academy of Agricultural Sciences, Hangzhou, 310008, China [3] Key Laboratory of Integrated Management of Crop Diseases and Pests (Ministry of Education), College of Plant Protection, Nanjing Agricultural University, Nanjing, 210095, China.

ABSTRACT
Insect chemosensory proteins (CSPs) are a family of small soluble proteins. To date, their physiological functions in insect olfaction remain largely controversial in comparison to odorant binding proteins (OBPs). In present study, we reported the antenna specific expression of three CSPs (AlinCSP4-6) from Adelphocoris lineolatus, their distinct chemosensillum distribution as well as ligand binding capability thus providing the evidence for the possible roles that they could play in semiochemical detection of the plant bug A. lineolatus. The results of qRT-PCR and western blot assay clearly showed that all of these three CSPs are highly expressed in the adult antennae, the olfactory organ of insects. Further cellular investigation of their immunolocalization revealed their dynamic protein expression profiles among different types of antennal sensilla. In a fluorescence competitive binding assay, the selective ligand binding was observed for AlinCSP4-6. In ad`dition, a cooperative interaction was observed between two co-expressed CSPs resulting in an increase of the binding affinities by a mixture of AlinCSP5 and AlinCSP6 to terpenoids which do not bind to individual CSPs. These findings in combination with our previous data for AlinCSP1-3 indicate a possible functional differentiation of CSPs in the A. lineolatus olfactory system.

No MeSH data available.