期刊
INSECT BIOCHEMISTRY AND MOLECULAR BIOLOGY
卷 140, 期 -, 页码 -出版社
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.ibmb.2021.103697
关键词
Bactrocera dorsalis; sNPFR; CRISPR; Cas9; Olfaction sensitivity; Foraging
资金
- National Natural Science Foundation of China [32072491, 31772233]
- National Key R&D Program of China [2021YFC2600100]
- Chongqing Natural Science Foundation [cstc2019jcyj-zdxmX0034]
- 111 Project [B18044]
- Chongqing Young Talents Support Program
- China Agriculture Research System of MOF
- China Agriculture Research System of MARA
By using the CRISPR/Cas9 system, a null sNPFR mutant was successfully generated to investigate its physiological roles in the oriental fruit fly. The mutants showed significant differences in foraging behavior and responses to odorants, as well as inhibition of several olfactory receptor genes. The data suggest that sNPFR plays a regulatory role in olfaction-mediated foraging behavior by interacting with specific cells and odorant receptor neurons.
The short neuropeptide F (sNPF) signaling system, consisting of sNPF and its receptor (sNPFR), influences many physiological processes in insects, including feeding, growth and olfactory memory. We previously showed that sNPF regulates olfactory sensitivity in the oriental fruit fly Bactrocera dorsalis (Hendel) during starvation. However, the functional analysis of sNPFR is constrained by the failure of RNA interference in this species. Here, we generated a null sNPFR mutant using the CRISPR/Cas9 system to investigate the physiological roles of this receptor in more detail. G0 adults were produced at a frequency of 60.8%, and sNPFR-/- mutants were obtained after several generations of backcrossing followed by self-crossing among heterozygous flies. We found that the mutants were significantly less successful at foraging for certain foods and showed increased foraging latency. Electroantennogram (EAG) assays indicated that the mutants had significantly lower electrophysiological responses to three tested odorants. Furthermore, qPCR data revealed the inhibition of several olfactory receptor genes, including Orco. Immunohistochemistry showed that BdsNPFR was localized in cells under the sensillum on the antennae. Based on their shape and size, the BdsNPFR+ cells differ from odorant receptor neurons (ORNs), which were labeled using a Drosophila melanogaster Orco antibody. Our data suggest that sNPFR regulates olfaction-mediated foraging behavior by mediating interactions between BdsNPFR+ cells and selected ORNs.
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