Molecular cloning, mRNA expression and biological activity of the pheromone biosynthesis activating neuropeptide (PBAN) from the European corn borer, Ostrinia nubilalis

Abstract

Pheromone biosynthesis activating neuropeptide (PBAN) is a member of the pyrokinin (FXPRLamide) insect neuropeptides. Here, we report the cloning of the gene Ostnu-PBAN from the E and Z pheromone strains of the European corn borer (ECB), Ostrinia nubilalis (Lepidoptera: Crambidae), a major pest of maize. The Ostnu-PBAN genomic sequence is > 5 kb in length and consists of six exons. The deduced amino acid sequence revealed a 200-residue precursor protein including a signal peptide, a 24-amino acid (aa) diapause hormone, a 37-aa PBAN and three other FXPRLamide neuropeptides. Our in vivo assays suggest that the 37-aa synthetic Ostnu-PBAN is hormonally active in the pheromone gland. It restores sex pheromone production to normal levels in mated females and decapitated virgins of both E and Z cultures. The results of a real-time PCR analysis indicated that Ostnu-PBAN mRNA levels reached a plateau in the brain-suboesophageal ganglion complexes 1 day after eclosion, and mating did not affect the mRNA expression. Three size classes of Ostnu-PBAN mRNA (1.9, 2.0 and 2.1 kb) were obtained, differing only in the length of the 3' untranslated region. However, there was no correlation between sequence divergence and the pheromone composition, voltinism or geographical origin (Hungary, Slovenia, Sweden, Turkey) of ECB moths.

Sex-specific spatial and temporal gene expressions of Pheromone biosynthesis activating neuropeptide (PBAN) and binding proteins (PBP/OBP) in Spoladea recurvalis

“…Notably, the cDNA encoding all five neuropeptides (Spre-DH homologue, Spre-α-NP, Spre-β-NP, Spre-PBAN, and Spre-γ-NP) in this study were identical to the members of Lepidoptera 17,18,27 . The presence of 6 exons interrupted by five introns of DH-PBAN has been reported for other genera, including Marvi-DH-PBAN, since these gene products were the precursor of sex pheromone biosynthesis activating neuropeptide 16,18 (Fig. 1).…”

“…The endoproteolytic cleavage, and glycine usually provide the amino group for amidation (Fig. 1) 38–40 , as reported for other DH-PBAN or PBAN that shared conserved pentapeptide sequence FXPR/KL at the C-termini 16–18 . Thus, amidation and physiological activity of DH and PBAN were essential, mainly for the members of Lepidoptera, Hymenoptera, Diptera, and Coleoptera 41 .…”

“…However, a variant C-terminal PBAN sequence showed asparagine substituted with serine in M. vitrata , revealing that this substitution was not commonly found in Pyraloidea super family (including this current study) (Fig. 1), which was associated with the pheromonotropic activity 16,18 (Fig. 1).…”

“…In Helicoverpa zea , a 33 amino acid C-terminal amidated peptide of PBAN was first identified 8 , whereas the determination of its amino acid sequence was done by direct isolation and purification from the Bombyx mori ’s head 14 . Later, PBAN encoding genes were identified in Aedes aegypti 15 , Ostrinia nubilalis 16 , Chlumentia transversa 17 , Maruca vitrata 18 , Manduca sexta 19 , and B. mori 20 . As a result, DH-PBAN proteins have been reported for more than 30 insect species spanning four orders including Lepidoptera and Hymenoptera 21 .…”

Male accessory gland factors increased sex pheromone titres in the glands ofSpodoptera liturafemale moths by inhibiting calling behavior

“…Molecular studies revealed that 64 sex pheromone production in female moths is regulated by the Pheromone 65 Biosynthesis Activating Neuropeptide (PBAN) (Raina & Kempe, 1990). Mating 66 seems will not affect the the mRNA expression of PBAN (Fodor et al, 2017), while 67 may inhibit the release of PBAN to the hemolymph and will result in a low 68 pheromone titre in the PG (Nagalakshimi et al, 2007). Nevertheless, the possible 69 links between MAG factors and PBAN are still unclear.…”