Identification and Gene Expression Analysis of the Pheromone Biosynthesis Activating Neuropeptide Receptor (PBANR) From the Ostrinia furnacalis (Lepidoptera: Pyralidae).

Pheromonal communication is important in insect mate finding and reproduction. Identifying components of pest insect pheromone system is a first step to disrupt pest insect reproduction. In this study, we identified and cloned the pheromone biosynthesis activating neuropeptide receptor (PBANR) from the Asian corn borer, Ostrinia furnacalis (Guenée) (Lepidoptera: Pyralidae), which is one of the most damaging pests of corn and other crops in parts of Asia and Australia. The O. furnacalis PBANR (OstfuPBANR) gene has an ORF of 1,086 bp and encoded 362 amino acids with seven transmembrane domains and had a high sequence identity to known lepidopteran PBANRs. Expression analysis showed that OstfuPBANR was highly expressed in the pheromone glands compared with other tissues, consistent with other studies. Interestingly, OstfuPBANR was expressed higher in the larval stages compared to the pupal or adult stages, suggesting that OstfuPBANR may have broad functions in larva beyond adult pheromone synthesis.

Luo M ，Zhou XC ，Wang Z ，Chen JX ，Chung H ，Wei HY ... -  《JOURNAL OF INSECT SCIENCE》

RNA interference of pheromone biosynthesis-activating neuropeptide receptor suppresses mating behavior by inhibiting sex pheromone production in Plutella xylostella (L.).

Sex pheromone production is regulated by pheromone biosynthesis-activating neuropeptide (PBAN) in many lepidopteran species. We cloned a PBAN receptor (Plx-PBANr) gene from the female pheromone gland of the diamondback moth, Plutella xylostella (L.). Plx-PBANr encodes 338 amino acids and has conserved structural motifs implicating in promoting G protein coupling and tyrosine-based sorting signaling along with seven transmembrane domains, indicating a typical G protein-coupled receptor. The expression of Plx-PBANr was found only in the pheromone gland of female adults among examined tissues and developmental stages. Heterologous expression in human uterus cervical cancer cells revealed that Plx-PBANr induced significant calcium elevation when challenged with Plx-PBAN. Female P. xylostella injected with double-stranded RNA specific to Plx-PBANr showed suppression of the receptor gene expression and exhibited significant reduction in pheromone biosynthesis, which resulted in loss of male attractiveness. Taken together, the identified PBAN receptor is functional in PBAN signaling via calcium secondary messenger, which leads to activation of pheromone biosynthesis and male attraction.

Lee DW ，Shrestha S ，Kim AY ，Park SJ ，Yang CY ，Kim Y ，Koh YH ... -  《-》

Expression of pheromone biosynthesis activating neuropeptide and its receptor (PBANR) mRNA in adult female Spodoptera exigua (Lepidoptera: Noctuidae).

The full-length cDNA of pheromone biosynthesis activating neuropeptide receptor (PBANR) was cloned from the beet armyworm, Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae); it included an open reading frame of 1,053 bp encoding 350 amino acids. The PBANR of S. exigua (SePBANR) was structurally characteristic of G protein-coupled receptor and its amino acid sequence shared 98% identity with the PBANR of Spodoptera littoralis. Both pheromone biosynthesis activating neuropeptide (PBAN) and PBANR mRNA abundance were measured in the brain-subesophageal ganglion complex, pheromone gland, ventral nerve cord, and ovary of S. exigua female moths by real-time RT-PCR. The abundance of PBAN mRNA in brain-subesophageal ganglion complex and PBANR mRNA in pheromone gland was significantly greater compared to other tissues, suggesting that the ligand-receptor relationship of PBAN and PBANR exists quantitatively in S. exigua. Both PBAN and PBANR expression displayed a remarkable diurnal rhythm, for they were low and stable during the photophase (07:00-21:00) and increased markedly during the scotophase (with a maximum abundance at 23:30) in 3-day-old female moths. The abundance of PBAN and PBANR increased steadily from the 1st day to the 5th day of the adult female life. The pattern of both diurnal and daily expression of PBAN and PBANR mRNA were coincident with enhanced capacity of sex pheromone release and mating of S. exigua moths during the same period. We infer from these results that pheromone biosynthesis and release in S. exigua is regulated by PBAN via up-regulating synthesis.

Cheng Y ，Luo L ，Jiang X ，Zhang L ，Niu C ... -  《-》

Identification and functional characterization of the pheromone biosynthesis activating neuropeptide receptor isoforms from Mamestra brassicae.

In most moth species, including Mamestra brassicae, pheromone biosynthesis activating neuropeptide (PBAN) regulates pheromone production. Generally, PBAN acts directly on the pheromone gland (PG) cells via its specific G protein-coupled receptor (i.e. PBANR) with Ca2+ as a second messenger. In this study, we identified cDNAs encoding three variants (A, B and C) of the M. brassicae PBANR (Mambr-PBANR). The full-length coding sequences were transiently expressed in cultured Trichoplusia ni cells and Sf9 cells for functional characterization. All three isoforms dose-dependently mobilized extracellular Ca2+ in response to PBAN analogs with Mambr-PBANR-C exhibiting the greatest sensitivity. Fluorescent confocal microscopy imaging studies demonstrated binding of a rhodamine red-labeled ligand (RR10CPBAN) to all three Mambr-PBANR isoforms. RR10CPBAN binding did not trigger ligand-induced internalization in cells expressing PBANR-A, but did in cells expressing the PBANR-B and -C isoforms. Furthermore, activation of the PBANR-B and -C isoforms with the 18 amino acid Mambr-pheromonotropin resulted in co-localization with a Drosophila melanogaster arrestin homolog (Kurtz), whereas stimulation with an unrelated peptide had no effect. PCR-based profiling of the three transcripts revealed a basal level of expression throughout development with a dramatic increase in PG transcripts from the day of adult emergence with PBANR-C being the most abundant.

Fodor J ，Hull JJ ，Köblös G ，Jacquin-Joly E ，Szlanka T ，Fónagy A ... -  《-》

Transcriptome Analysis of Ostrinia furnacalis Female Pheromone Gland: Esters Biosynthesis and Requirement for Mating Success.

Female moths use sex pheromones to attract males, and corresponding regulatory mechanism underlying sex pheromone biosynthesis is species-dependent. However, the detailed mechanism involved in sex pheromone biosynthesis in Ostrinia furnacalis has not yet been fully addressed. In the present study, transcriptome sequencing of O. furnacalis pheromone glands screened a serials of candidate genes involved in sex pheromone biosynthesis. Our analysis showed that sex pheromone release in O. furnacalis females arrives its peak at the 2nd scotophase, consistent with its mating behavior. Pheromone biosynthesis-activating neuropeptide (PBAN) was confirmed to regulate sex pheromone biosynthesis, and Ca2+ is the secondary messenger of PBAN signaling in O. furnacalis. The functional analysis of candidate genes demonstrated that the decreased mRNA levels or activities of calcineurin (CaN) and acetyl-CoA carboxylase (ACC) led to significant decrease in sex pheromone production and female capability to attract males, as demonstrated by RNAi-mediated knockdown and pharmacological inhibitor assay. Most importantly, the activities of CaN and ACC depend on the activation of PBAN/PBANR/Ca2+. Furthermore, fatty-acyl reductase 14 was involved in PBAN-mediated sex pheromone biosynthesis. Altogether, our results demonstrated that PBAN regulates sex pheromone biosynthesis through PBANR/Ca2+/CaN/ACC pathway to promote sex pheromone biosynthesis in O. furnacalis and provided a reference for non-model organism to study neuropeptide signal transduction.

Yao S ，Zhou S ，Li X ，Liu X ，Zhao W ，Wei J ，Du M ，An S ... -  《Frontiers in Endocrinology》