Identification and expression profiles of novel odorant binding proteins and functional analysis of OBP99a in Bactrocera dorsalis.

Odorant-binding proteins (OBPs) in insects are essential for mating and oviposition host selection. How these OBPs respond to different hosts at the mRNA level and their effects on behavior remain poorly characterized. The oriental fruit fly Bactrocera dorsalis is a highly invasive agricultural pest with an extremely broad host range and high fecundity. Based on our previously constructed B. dorsalis transcriptome, six OBPs that were differentially expressed during three different physiological adult stages were identified. A phylogenetic tree was constructed to illustrate the relationships of these six OBPs with OBP sequences from three other dipteran species (Drosophila melanogaster, Anopheles gambiae, and Ceratitis capitata). The spatiotemporal expression profiles of the six OBPs were analyzed using quantitative real-time PCR. Our results revealed that OBP19c, OBP44a, OBP99a, and OBP99d were abundantly expressed from the prepupa stage to the adult stage, and most of the OBPs were mainly expressed in the head, wings, and antennae. The expression levels of these OBPs were upregulated when female flies were exposed to their preferred hosts. Silencing OBP99a resulted fewer eggs being laid compared with the control group when the females were exposed to their preferred host, that is, banana, whereas more eggs were laid when a non-preferred host, that is, tomato, was used. Furthermore, silencing OBP99a led to sexually dimorphic mating behavior. dsOBP99a-injected males dramatically reduced courtship, whereas enhanced courtship was observed in the treated females. These data indicate that OBPs may participate in different biological processes of B. dorsalis. Our study will provide insight into the molecular mechanism of chemoreception and help develop ecologically friendly pest-control strategies.

RNA sequencing to characterize transcriptional changes of sexual maturation and mating in the female oriental fruit fly Bactrocera dorsalis.

BACKGROUND: Female reproductive potential plays a significant role in the survival and stability of species, and sexual maturation and mating processes are crucial. However, our knowledge of the reproductive genes involved in sexual maturation and mating has been largely limited to model organisms. The oriental fruit fly Bactrocera dorsalis is a highly invasive agricultural pest, known to cause major economic losses; thus, it is of great value to understand the transcriptional changes involved in sexual maturation and mating processes as well as the related genes. Here, we used a high-throughput sequencing method to identify multiple genes potentially involved in sexual maturation and mating in female B. dorsalis. RESULTS: We sequenced 39,999 unique genes with an average length of 883 bp. In total, 3264 differentially expressed genes (DEGs) were detected between mature virgin and immature Bactrocera dorsalis libraries, whereas only 83 DEGs were identified between flies that had mated or were mature virgins. These DEGs were functionally annotated using the GO and KEGG pathway annotation tools. Results showed that the main GO terms associated with the DEGs from the mature virgin vs. immature groups were primarily assigned to the metabolic and developmental processes, which we focused on, whereas those from the mated vs. mature virgin group largely belonged to the response to stimulus and immune system processes. Additionally, we identified multiple DEGs during sexual maturation that are involved in reproduction, and expression pattern analysis revealed that the majority DEGs detected were highly enriched in those linked to the ovaries or fat bodies. Several mating responsive genes differentially expressed after mating were also identified, and all antimicrobial peptides detected were highly enriched in fat body and significantly up-regulated approximately 2- to 10-fold at 24 h after mating. CONCLUSION: This study supplied female reproductive genes involved in sexual maturation and the post-mating response in B. dorsalis, based on RNA-seq. Our data will facilitate molecular research related to reproduction and provide abundant target genes for effective control of this agricultural pest.