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      Expression and functional characterization of odorant‐binding protein 2 in the predatory mite Neoseiulus barkeri

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          Abstract

          Olfaction plays a crucial role for arthropods in foraging, mating, and oviposition. The odorant‐binding protein (OBP) gene is considered one of the most important olfactory genes. However, little is known about its functions in predatory mites. Here, we used Neoseiulus barkeri, an important commercialized natural pest control, to explore the chemosensory characteristics of OBP. In this study, N. barkeri was attracted by methyl salicylate (MeSA) and showed higher crawling speeds under MeSA treatment. Then, we identified and cloned an OBP gene named Nbarobp2 and analyzed its expression profiles in the predatory mite. Nbarobp2 was 663 bp, was highly expressed in larval and nymphal stages, and was significantly upregulated in N. barkeri under MeSA treatment. Nbarobp2 encoded 202 amino acid residues with a molecular weight of 23 kDa (after removing the signal peptide). Sequence comparisons revealed that the OBPs in Arachnida shared 6 conserved cysteine sites, but were distinguishable from the OBPs of Insecta on the phylogenetic tree. RNA interference, Western blotting, and binding affinity assays further proved that Nbarobp2 was involved in volatile perception in predatory mites. This study shed light on the functional characteristics of OBPs in predatory mites, providing a new insight for better biological control.

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          Odorant reception in insects: roles of receptors, binding proteins, and degrading enzymes.

          Our knowledge of the molecular basis of odorant reception in insects has grown exponentially over the past decade. Odorant receptors (ORs) from moths, fruit flies, mosquitoes, and the honey bees have been deorphanized, odorant-degrading enzymes (ODEs) have been isolated, and the functions of odorant-binding proteins (OBPs) have been unveiled. OBPs contribute to the sensitivity of the olfactory system by transporting odorants through the sensillar lymph, but there are competing hypotheses on how they act at the end of the journey. A few ODEs that have been demonstrated to degrade odorants rapidly may act in signal inactivation alone or in combination with other molecular traps. Although ORs in Drosophila melanogaster respond to multiple odorants and seem to work in combinatorial code involving both periphery and antennal lobes, reception of sex pheromones by moth ORs suggests that their labeled lines rely heavily on selectivity at the periphery.
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            Pheromone binding and inactivation by moth antennae

            The antennae of male silk moths are extremely sensitive to the female sex pheromone such that a male moth can find a female up to 4.5 km away. This remarkable sensitivity is due to both the morphological and biochemical design of these antennae. Along the branches of the plumose antennae are the sensilla trichodea, each consisting of a hollow cuticular hair containing two unbranched dendrites bathed in a fluid, the receptor lymph ,3. The dendrites and receptor lymph are isolated from the haemolymph by a barrier of epidermal cells which secreted the cuticular hair. Pheromone molecules are thought to diffuse down 100 A-wide pore tubules through the cuticular wall and across the receptor lymph space to receptors located in the dendritic membrane. To prevent the accumulation of residual stimulant and hence sensory adaptation, the pheromone molecules are subsequently inactivated in an apparent two-step process of rapid 'early inactivation' followed by much slower enzymatic degradation. The biochemistry involved in this sequence of events is largely unknown. We report here the identification of three proteins which interact with the pheromone of the wild silk moth Antheraea polyphemus: a pheromone-binding protein and a pheromone-degrading esterase, both uniquely located in the pheromone-sensitive sensilla; and a second esterase common to all cuticular tissues except the sensilla.
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              Discrimination of alarm pheromone (E)-beta-farnesene by aphid odorant-binding proteins.

              OBPs have been recently demonstrated to be required for odour perception in insects and directly involved in odour discrimination. In aphids they might represent new interesting targets for the control of their population in agriculture. Based on sequence information available in the EST database, we have cloned four genes encoding odorant-binding proteins (OBP) in Acyrthosiphon pisum and homologous genes in other aphid species. Unlike OBPs from other orders of insects, that are greatly divergent, in aphids these proteins have been found to be highly conserved, with differences between species limited to only few amino acid substitutions. On the contrary, similarities between OBP sequences of the same species are poor with 31% or less of identical amino acids. Three selected OBPs (OBP1, OBP3 and OBP8) have been expressed in bacteria and purified. Ligand-binding experiments have shown similar behaviour of the three proteins towards several organic compounds, but also some significant selectivities. In particular, (E)-beta-farnesene, the alarm pheromone and its related compound farnesol exhibited good affinity to OBP3, but did not bind the other two proteins. We suggest that OBP3 could mediate response of aphids to the alarm pheromone.
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                Author and article information

                Contributors
                Journal
                Insect Science
                Insect Science
                Wiley
                1672-9609
                1744-7917
                October 2023
                January 09 2023
                October 2023
                : 30
                : 5
                : 1493-1506
                Affiliations
                [1 ] Key Laboratory of Entomology and Pest Control Engineering, College of Plant Protection Southwest University Beibei District Chongqing 400715 China
                Article
                10.1111/1744-7917.13156
                36458978
                e779d17b-d98a-4556-bad8-bae081ac6b72
                © 2023

                http://onlinelibrary.wiley.com/termsAndConditions#vor

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