Molecular and in vitro biochemical assessment of chemosensory protein 10 from brown planthopper Nilaparvata lugens at acidic pH

Chemosensory proteins(CSPs)are important molecular components of the insect olfactory system,which are involved in capturing,binding,and transporting hydrophobic odour molecules across the sensillum in sensillar lymph in regulating insect behavior.This protein family(CSPs)is also involved in many other systems that are not linked to olfactory receptors in olfactory sensilla.The brown planthopper(BPH)is a monophagous pest of rice that causes damage by sucking phloem sap and transmitting a number of diseases caused by viruses.In this study,fluorescence competitive binding assay and fluorescence quenching assay at acidic p H were performed as well as homology modelling to describe the binding affinity of Nlug CSP10.Fluorescence competitive binding assay(FCBA)demonstrated that Nlug CSP10 bound strongly to nonadecane,farnesene,and 2-tridecanone at acidic p H.The results of FCBA indicated that Nlug CSP10 bound different ligands at the physiological p H(5.0)of the bulk sensillum lymph.Fluorescence quenching assay demonstrated that Nlug CSP10 generated a stable complex with 2-tridecanone,while two ligands nonadecane and farnesene collided due to molecular collisions.The interaction of selected ligands with the modelled structure of Nlug CSP10 was also analyzed,which found the key amino acids(Gln23,Gln24,Gln25,Asn27,Met33,Ser34,Ile35,Tyr36,Asn42,Met43,Val45,Asn46,Asn93,Arg96,Ala97,Lys99,and Ala100)in Nlug CSP10 that were involved in binding of volatile compounds.The present study contributes to the binding profile of Nlug CSP10 that promotes the development of behaviorally active ligands based on BPH olfactory system.

The role of chemosensory protein 10 in the detection of behaviorally active compounds in brown planthopper, Nilaparvata lugens

Chemosensory proteins(CSPs)play important roles in insects’chemoreception,although their specific functional roles have not been fully elucidated.In this study,we conducted the developmental expression patterns and competitive binding assay as well as knock‐down assay by RNA interference both in vitro and in vivo to reveal the function of NlugCSP10 from the brown planthopper(BPH),Nilaparvata lugens(Stål),a major pest in rice plants.The results showed that NlugCSP10 messenger RNA was significantly higher in males than in females and correlated to gender,development and wing forms.The fluorescence binding assays revealed that NlugCSP10 exhibited the highest binding affinity with cis‐3‐hexenyl acetate,eicosane,and(+)‐β‐pinene.Behavioral assay revealed that eicosane displayed attractant activity,while cis‐3‐hexenyl acetate,similar to(+)‐β‐pinene significantly repelled N.lugens adults.Silencing of NlugCSP10,which is responsible for cis‐3‐hexenyl acetate binding,significantly disrupted cis‐3‐hexenyl acetate communication.Overall,findings of the present study showed that NlugCSP10 could selectively interrelate with numerous volatiles emitted from host plants and these ligands could be designated to develop slow‐release mediators that attract/repel N.lugens and subsequently improve the exploration of plans to control this insect pest.

Appraisal of MsepCSP14 for chemosensory functions in Mythimna separata

Chemosensory proteins (CSPs) have great contributions in performing diverse functions in insects. However, physiological appraisal of chemosensory protein genes still remains elusive in insects. We studied expression patterns and binding affinities of MsepCSP14, a chemosensory protein, in Mythimna separata. The distinct functions of MsepCSP14 were validated by employing different molecular techniques. The MsepCSP14 had high resemblance of sequence with chemosensory proteins of other insect family members. The MsepCSP14 expression was higher in antennal tissues of females than other tissues. Fluorescence binding assay validated that binding of nine out of 21 ligands to MsepCSP14 was higher at pH 7.4 than at pH 5.0. Three dimensional modeling (3D) and docking analysis predicted that amino acid residues of MsepCSP14 were involved in binding of compounds, and behavior assay displayed that adults of M. separata considerably responded to four volatiles from compounds demonstrating strong binding ability to MsepCSP14. Results of the present study suggest that MsepCSP14 is likely to mediate chemosensory functions in M. separata.