Two general odorant binding proteins display high bindings to both host plant volatiles and sex pheromones in a pyralid moth Chilo suppressalis (Lepidoptera: Pyralidae)

Khuhro, Sajjad Ali; Liao, Hui; Dong, Xiao-Tong; Yu, Qi; Yan, Qi; Dong, Shuang-Lin

doi:10.1016/j.aspen.2017.02.015

Cited by 47 publications

(55 citation statements)

References 54 publications

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“…This might be due to compensation by other PBPs and even OBPs and CSPs. In vitro ligand binding assays suggested that general odorant binding proteins (GOBPs, particularly GOBP2) may function to transport sex pheromones in C. suppressalis (Gong et al, 2009;Khuhro et al, 2017) and some other moths (Jacquinjoly et al, 2000;Zhou et al, 2009;He et al, 2010;Liu et al, 2012b); an antenna-predominant and male-biased CSP19 in Sesamian inferens showed high binding affinities to female sex pheromones with Ki values comparable to that of PBPs, suggesting its involvement in sex pheromone perception (Zhang et al, 2014). Furthermore, interactions among OBPs in ligand binding were found in Holotrichia oblita (Wang et al, 2013a).…”

Section: Discussionmentioning

confidence: 99%

CRISPR/Cas9‐mediated PBP1 and PBP3 mutagenesis induced significant reduction in electrophysiological response to sex pheromones in male Chilo suppressalis

et al. 2017

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Pheromone-binding proteins (PBPs) are thought to bind and transport sex pheromones onto the olfactory receptors on the dendrite membrane of olfactory neurons, and thus play a vital role in sex pheromone perception. However, the function of PBPs has rarely been demonstrated in vivo. In this study, two PBPs (PBP1 and PBP3) of Chilo suppressalis, one of the most notorious pyralid pests, were in vivo functionally characterized using insects with the PBP gene knocked out by the CRISPR/Cas9 system. First, through direct injection of PBP-single guide RNA (sgRNA)/Cas9 messenger RNA into newly laid eggs, a high rate of target-gene editing (checked with polled eggs) was induced at 24 h after injection, 21.3% for PBP1-sgRNA injected eggs and 19.5% for PBP3-sgRNA injected eggs. Second, by an in-crossing strategy, insects with mutant PBP1 or PBP3 (both with a premature stop codon) were screened, and homozygous mutants were obtained in the G3 generation. Third, the mutant insects were measured for electroantennogram (EAG) response to female sex pheromones. As a result, both PBP mutant males displayed significant reduction in EAG response, and this reduction in PBP1 mutants was higher than that in PBP3 mutants, indicating a more important role of PBP1. Finally, the relative importance of two PBPs and the possible off target effect induced by sgRNA-injection are discussed. Taken together, our study provides a deeper insight into the function of and interaction between different PBP genes in sex pheromone perception of C. suppressalis, as well as a valuable reference in methodology for gene functional study in other genes and other moth species.

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Section: Discussionmentioning

confidence: 99%

CRISPR/Cas9‐mediated PBP1 and PBP3 mutagenesis induced significant reduction in electrophysiological response to sex pheromones in male Chilo suppressalis

et al. 2017

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“…The collected tissue samples (200 antennae, 40 heads, 80 legs, and 20 abdomens of each sex) were separately placed in liquid nitrogen and stored at −80°C until use. Total RNA isolation and cDNA synthesis were performed as our reported protocols ( Khuhro et al 2017a , b ).…”

Section: Methodsmentioning

confidence: 99%

“…The recombinant proteins were expressed by bacterial expression system and protein purification was accomplished by previously described protocols ( Khuhro et al 2017a , b ). To avoid the possible effects of His-tags on the structures of CSPs, tags were removed by the treatment with a cleaving enzyme (enterokinase), and stored at – 80°C for further use.…”

Section: Methodsmentioning

confidence: 99%

“…The binding assays were conducted using previously reported protocols ( Liu et al 2012 ; Liu et al 2013 ; Khuhro et al 2017a , b ) on a Spectra Max M5 Fluorescence Spectrophotometer (Molecular Devices, California, USA) with Greiner Microlon 96-well plates (Greiner Bio-One, Frickenhausen, Germany). The solutions were excited at 337 nm and emission spectra were recorded between 400 and 460 nm.…”

Section: Methodsmentioning

confidence: 99%

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Expression Profile and Functional Characterization Suggesting the Involvement of Three Chemosensory Proteins in Perception of Host Plant Volatiles in Chilo suppressalis (Lepidoptera: Pyralidae)

Khuhro

Yan

Liao

et al. 2018

Journal of Insect Science

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The high sensitivity of the olfactory system is essential for feeding and oviposition in moth insects, and some chemosensory proteins (CSPs) are thought to play roles in this system by binding and carrying hydrophobic odorants across the aqueous sensillar lymph. In this study, to identify the olfactory CSPs from a repertoire of 21 CSP members in the notorious rice pest Chilo suppressalis (Walker) (Lepidoptera: Pyralidae), tissue expression patterns were firstly examined by quantitative real-time polymerase chain reaction (qPCR). It showed that CSP2 was antennae specific and seven more CSPs (CSP1, 3, 4, 6, 15, 16, and 17) were antennae biased in expression, suggesting their olfactory roles; while other CSPs were multiple-tissue expressed and non-antennae biased, suggesting other functions for these genes. To further determine the ligand binding specificity, three putative olfactory genes (CSP1–3) were expressed in Escherichia coli cells, and binding affinity of these three recombinant CSP proteins were measured for 35 plant volatiles by the ligand binding assays. CSP1 and CSP2 exhibited high binding affinities (Ki ≤ 10.00 µM) for four (2-tridecanone, benzaldehyde, laurinaldehyde and 2-pentadecanone) and two (2-heptanol and (+)-cedrol) host plant volatiles, respectively; the three CSPs also showed moderate binding affinity (Ki = 10.01–20.00 µM) for 16 plant volatiles. Our study suggests that the three CSPs play essential roles in the perception of host plant volatiles, providing bases for the elucidation of olfactory mechanisms in this important pyralid pest.

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“…They bound to most alcohols, esters and ketones tested, but not to any of the alkanes or the aldehyde (Table ), suggesting the importance of functional groups of odorant molecules when they interact with SmosOBPs. Similar results have also been found in OBPs of many other insects, such as OBP8, 11 and 15 in Grapholita molesta (Li et al ., ), OBP1 in Adelphocoris lineolatus (Gu et al ., ), OBP1 and 2 in Culex quinquefasciatus (Yin et al ., ) and Chilo suppressalis (Khuhro et al ., ). Consistent with the binding assay results, most of alcohols and esters elicited significant EAG responses from female adults, whereas all alkanes and the aldehyde tested aroused no or weak responses (Fig.…”

Section: Discussionmentioning

confidence: 97%

Molecular and functional characterization of three odorant‐binding proteins from the wheat blossom midge, Sitodiplosis mosellana

et al. 2019

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Sitodiplosis mosellana, a periodic but devastating wheat pest, relies on wheat spike volatiles as a cue in selecting hosts for oviposition. Insect odorant-binding proteins (OBPs) are thought to play essential roles in filtering, binding and transporting hydrophobic odorant molecules to specific receptors. To date, the molecular mechanisms underlying S. mosellana olfaction are poorly understood. Here, three S. mosellana antenna-specific OBP genes, SmosOBP11, 16 and 21, were cloned and bacterially expressed. Binding properties of the recombinant proteins to 28 volatiles emitted from wheat spikes were investigated using fluorescence competitive binding assays. Sequence analysis suggested that these SmosOBPs belong to the Classic OBP subfamily. Ligand-binding analysis showed that all three SmosOBPs preferentially bound alcohol, ester and ketone compounds, and SmosOBP11 and 16 also selectively bound terpenoid compounds. In particular, the three SmosOBPs had high binding affinities (K i < 20 μmol/L) to 3-hexanol and cis-3-hexenylacetate that elicited strong electroantennogram (EAG) response from female antennae. In addition, SmosOBP11 displayed significantly higher binding (K i < 8 μmol/L) than SmosOBP16 and 21 to 1-octen-3-ol, D-panthenol, α-pinene and heptyl acetate which elicited significant EAG response, suggesting that SmosOBP11 plays a major role in recognition and transportation of these volatiles. These findings have provided important insight into the molecular mechanism by which S. mosellana specifically recognizes plant volatiles for host selection, and have facilitated identification of effective volatile attractants that are potentially useful for pest monitoring and trapping.

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Two general odorant binding proteins display high bindings to both host plant volatiles and sex pheromones in a pyralid moth Chilo suppressalis (Lepidoptera: Pyralidae)

Cited by 47 publications

References 54 publications

CRISPR/Cas9‐mediated PBP1 and PBP3 mutagenesis induced significant reduction in electrophysiological response to sex pheromones in male Chilo suppressalis

CRISPR/Cas9‐mediated PBP1 and PBP3 mutagenesis induced significant reduction in electrophysiological response to sex pheromones in male Chilo suppressalis

Expression Profile and Functional Characterization Suggesting the Involvement of Three Chemosensory Proteins in Perception of Host Plant Volatiles in Chilo suppressalis (Lepidoptera: Pyralidae)

Molecular and functional characterization of three odorant‐binding proteins from the wheat blossom midge, Sitodiplosis mosellana

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