Cloning, expression and binding specificity analysis of odorant binding protein 3 of the lucerne plant bug, Adelphocoris lineolatus (Goeze)

Abstract

Credible evidence shows that odorant binding proteins (OBPs) are required for insect olfaction perception and play a key role in transporting hydrophobic odorants across the sensillum lymph to the olfactory receptors (ORs). In the present study, a novel OBP (AlinOBP3) gene from the lucerne plant bug, Adelphocoris lineolatus, was cloned and expressed. The expression pattern of AlinOBP3 was evaluated by qPCR, which indicated that AlinOBP3 was dominantly expressed in antennae. The binding properties of AlinOBP3 with 9 cotton volatiles and 5 sex pheromone analogs were measured by fluorescence competitive binding assays with the fluorescence probe 1-NPN. The results revealed that of 9 cotton volatiles, Myrcene, β-Ocimene and α-Phellandrene can bind with AlinOBP3. α-Phellandrene especially bound to AlinOBP3 with a high binding affinity, with a dissociation constant of 56.68 μmol/L. Of the 5 sex pheromone analogs, Hexyl butyrate had the strongest binding affinity with AlinOBP3, with a dissociation constant as 59.53 μmol/L. Butyl butyrate, trans-2-Hexenyl butyrate and Ethyl butyrate had medium binding affinities with AlinOBP3, with dissociation constants of 227.39, 108.77 and 143.47 μmol/L, respectively. The results suggest that AlinOBP3 might be a pheromone binding protein (PBP) with a dual-function for the perception of sex pheromones and plant volatiles. Adelphocoris lineolatus (Goeze), odorant binding protein, gene expression, Western blot, expression pattern, fluorescence competitive binding assays Citation: Gu S H, Sun Y, Ren L Y, et al. Cloning, expression and binding specificity analysis of odorant binding protein 3 of the lucerne plant bug, Adelphocoris lineolatus (Goeze).

Specific involvement of two amino acid residues in cis‐nerolidol binding to odorant‐binding protein 5 AlinOBP5 in the alfalfa plant bug, Adelphocoris lineolatus (Goeze)

“…). The inclusion body of AlinOBP5 was solubilized, refolded and purified by His‐Trap affinity columns (GE Healthcare, Waukesha, WI, USA), followed by removal of the His‐tag with enterokinase as described previously (Gu et al ., , ). The SDS‐PAGE analysis indicated that the molecular weight of the final purified AlinOBP5 without the His‐tag was about 13 kDa (Fig.…”

Cloning and expression profiling of odorant‐binding proteins in the tarnished plant bug, Lygus lineolaris

“…Furthermore, trans‐2‐hexenyl butyrate has been shown to elicit a strong antennal electrophysiological response in male L. lineolaris (Chinta et al ., ). AdlinOBP3, in contrast, was found to only weakly bind α‐phellandrene, a cotton volatile (Gu et al ., ). That study was constrained by a limited number of compounds assayed, which probably contributed to the poor determination of a bona fide AdlinOBP3 substrate.…”

Functional analysis of female‐biased odorant binding protein 6 for volatile and nonvolatile host compounds in Adelphocoris lineolatus (Goeze)

“…In A. lineolatus, 14 OBP genes have been identified (Gu et al, 2011b). The antennae-enriched OBPs, including AlinOBP1, 2, 3, 5 10 and 13, all displayed preferential binding properties to host plant volatiles and/or bugproduced compounds (Gu et al, 2010(Gu et al, , 2011aSun et al, 2013Sun et al, , 2014bWang et al, 2013). In particular, AlinOBP1 and AlinOBP13 are specifically expressed in the lymph of the multiporous sensilla trichodea, the basiconica and the spoke channel peg and function in exclusively olfactory roles in response to biologically relevant odorants (Gu et al, 2011c;Sun et al, 2014b).…”