Functional differentiation of three pheromone binding proteins in Orthaga achatina using mixed-type sex pheromones

Si, Yu-Xiao; Guo, Jin‐Meng; Liao, Hui; Liu, Yu; Ma, Yu; Zhu, Yu-Wei; Wei, Zhiqiang; Dong, Shuang-Lin; Yan, Qi

doi:10.1016/j.pestbp.2022.105097

Cited by 6 publications

(5 citation statements)

References 52 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Consistent with the fluorescence binding assays, the docking results showed lower binding energies between CbuqPBP2 with three tested compounds, suggesting strong binding strength. It was proved that PBPs commonly bind ligands in hydrophobic cavities, such as PBPs from Orthaga achatina [45]. In our study, it was also found that hydrophobic interactions were the prevailing forces within the binding cavities of CbuqPBP2.…”

Section: Discussionsupporting

confidence: 65%

Molecular and Functional Characterization of Pheromone Binding Protein 2 from Cyrtotrachelus buqueti (Coleoptera: Curculionidae)

Liu,

Wang,

Yang

et al. 2023

IJMS

View full text Add to dashboard Cite

Pheromone-binding proteins (PBPs) play important roles in binding and transporting sex pheromones. However, the PBP genes identified in coleopteran insects and their information sensing mechanism are largely unknown. Cyrtotrachelus buqueti (Coleoptera: Curculionidae) is a major insect pest of bamboo plantations. In this study, a novel PBP gene, CbuqPBP2, from C. buqueti was functionally characterized. CbuqPBP2 was more abundantly expressed in the antennae of both sexes than other body parts, and its expression level was significantly male-biased. Fluorescence competitive binding assays showed that CbuqPBP2 exhibited the strongest binding affinity to dibutyl phthalate (Ki = 6.32 μM), followed by styrene (Ki = 11.37 μM), among twelve C. buqueti volatiles. CbuqPBP2, on the other hand, showed high binding affinity to linalool (Ki = 10.55), the main volatile of host plant Neosinocalamus affinis. Furthermore, molecular docking also demonstrated the strong binding ability of CbuqPBP2 to dibutyl phthalate, styrene, and linalool, with binding energy values of −5.7, −6.6, and −6.0 kcal/mol, respectively, and hydrophobic interactions were the prevailing forces. The knockdown of CbuqPBP2 expression via RNA interference significantly reduced the electroantennography (EAG) responses of male adults to dibutyl phthalate and styrene. In conclusion, these results will be conducive to understanding the olfactory mechanisms of C. buqueti and promoting the development of novel strategies for controlling this insect pest.

show abstract

Section: Discussionsupporting

confidence: 65%

Molecular and Functional Characterization of Pheromone Binding Protein 2 from Cyrtotrachelus buqueti (Coleoptera: Curculionidae)

Liu,

Wang,

Yang

et al. 2023

IJMS

View full text Add to dashboard Cite

show abstract

“…The PCR product was purified from an agarose gel, ligated into the pEASY-Blunt 3 vector (TransGen Biotech, Beijing, China), and then transferred into Trans-T 1 cells. The expression and purification of recombinant proteins were conducted using previously described methods [ 18 ].…”

Section: Methodsmentioning

confidence: 99%

“…The qPCR reaction was performed in 20 μL: containing 10 μL of SYBR Green qPCR Master Mix, 0.4 μL of ROX Reference Dye II, 0.4 μL of each primer (10 μM), 1 μL of cDNA (10 ng/μL) template, and 7.8 μL of nuclease-free water. The reaction procedure was same as that used in previous studies [ 18 ]. GAPDH and β-actin ( Table S1 ) were used as the reference gene to normalize the target gene expression (GenBank KT361883) [ 20 ].…”

Section: Methodsmentioning

confidence: 99%

Identification and Functional Characterization of General Odorant Binding Proteins in Orthaga achatina

Wei

et al. 2023

Insects

Self Cite

View full text Add to dashboard Cite

The olfactory system in insects are crucial for recognition of host plants and oviposition sites. General odorant binding proteins (GOBPs) are thought to be involved in detecting odorants released by host plants. Orthaga achatina (Lepidoptera: Pyralidae) is one of the most serious pests of camphor trees, Cinnamomum camphora (L.) Presl, an important urban tree species in southern China. In this study, we study the GOBPs of O. achatina. Firstly, two full-length GOBP genes (OachGOBP1 and OachGOBP2) were successfully cloned according to transcriptome sequencing results, and real-time quantitative PCR measurements showed that both GOBP genes were specifically expressed in the antennae of both sexes, proposing their important roles in olfaction. Then, both GOBP genes were heterologous expressed in Escherichia coli and fluorescence competitive binding assays were conducted. The results showed that OachGOBP1 could bind Farnesol (Ki = 9.49 μM) and Z11-16: OH (Ki = 1.57 μM). OachGOBP2 has a high binding affinity with two camphor plant volatiles (Farnesol, Ki = 7.33 μM; α-Phellandrene, Ki = 8.71 μM) and two sex pheromone components (Z11-16: OAc, Ki = 2.84 μM; Z11-16: OH, Ki = 3.30 μM). These results indicate that OachGOBP1 and OachGOBP2 differ in terms of odorants and other ligands. Furthermore, key amino acid residues that bind to plant volatiles were identified in GOBPs using 3-D structure modeling and ligand molecular docking, predicting the interactions between the GOBPs and the host plant volatiles.

show abstract

“…The total reaction volume was 20 μL: containing 10 μL of 2× Cham Q Universal SYBR qPCR Master Mix, 0.4 μL of each primer (10 μM), 8.2 μL of ddH 2 O, and 1 μL of cDNA. The reaction procedure was same as a previous study Table S1) were used as the reference genes to normalize the target gene expression .…”

Section: Methodsmentioning

confidence: 99%

“…The reaction procedure was same as a previous study. 16 GAPDH and βactin (Table S1) were used as the reference genes to normalize the target gene expression. 17 Gene expression levels were analyzed using the 2 −ΔCT method, and data were analyzed by GraphPad Prism 8.0.…”

Section: Tissue Expression Profiles By Qpcrmentioning

confidence: 99%

The Odorant Receptor Recognizing Camphor in a Camphor Tree Specialist Orthaga achatina (Lepidoptera: Pyralidae)

Ma,

Yang,

et al. 2024

J. Agric. Food Chem.

Self Cite

View full text Add to dashboard Cite

Camphor has been used as an effective repellent and pesticide to stored products for a long history, but Orthaga achatina (Lepidoptera: Pyralidae) has evolved to specifically feed on the camphor tree Cinnamomum camphora. However, the behavioral response of O. achatina to camphor and the molecular basis of camphor perception are totally unknown. Here, we demonstrated that both male and female adults were behaviorally attracted to camphor, suggesting the adaptation of O. achatina to and utilization of camphor as a signal of C. camphora. Second, in 40 O. achatina OR genes obtained by analyzing antenna transcriptomes, only OachOR16/Orco significantly responded to camphor in the Xenopus oocyte system. Finally, by molecular docking analysis and site-directed mutagenesis, the Ser209 residue is confirmed to be essential for binding of the oachOR16 with camphor. This study not only reveals the camphor-based host plant choice and olfactory mechanisms of O. achatina but also provides a molecular target for screening more potential insect repellents.

show abstract

Functional differentiation of three pheromone binding proteins in Orthaga achatina using mixed-type sex pheromones

Cited by 6 publications

References 52 publications

Molecular and Functional Characterization of Pheromone Binding Protein 2 from Cyrtotrachelus buqueti (Coleoptera: Curculionidae)

Molecular and Functional Characterization of Pheromone Binding Protein 2 from Cyrtotrachelus buqueti (Coleoptera: Curculionidae)

Identification and Functional Characterization of General Odorant Binding Proteins in Orthaga achatina

The Odorant Receptor Recognizing Camphor in a Camphor Tree Specialist Orthaga achatina (Lepidoptera: Pyralidae)

Contact Info

Product

Resources

About