Hox genes, specifically the bithorax complex (ubx,abdominal-a, andabdominal-b), play a crucial role in specifying posterior abdominal development and serve as key regulators of germline gene development in insects. However, the function of the bithorax complex in the pine caterpillar moth,Dendrolimus punctatus, a major pine tree defoliator in China, remains largely unknown. Specifically,Abdominal-A(Abd-a) controls regional variation in abdominal segmentation in model insects such asDrosophilaandTribolium; however, its role inD. Punctatusremains unexplored. In this study, CRISPR/Cas9 was used to functionally characterizeAbd-ainD. punctatus. Two target sites were selected, and the genotypes and phenotypes of the G0 and G1 generations were evaluated. Our findings indicate that knocking outAbd-aled to an abnormality in the posterior segments A2–A7, as well as the loss of appendages, mainly prolegs, and affected the thoracic T3 segmentation as well as wing development. Moreover, mutation inAbd-aalso impacted anal and reproductive development. Taken together, these results demonstrate thatDpAbd-ais essential for embryonic and reproductive development inD. punctatusand could be a promising target for genetic control of this devastating conifer defoliator.
Jian Yang,Jia Wang,Xiaowei Peng,Chengfeng Lei,Xiulian Sun,Jia Hu
Dendrolimus punctatuscauses great damage to pine forests worldwide. Dendrolimus punctatus cypovirus 1 (DpCPV-1) is an important pathogen ofD. punctatus. However, the mechanism of DpCPV-1 cell entry has not been elucidated. In this study, we revealed that both GTase and MTase domains of VP3 (B-spike) and VP4 (A-spike) of DpCPV-1 interacted with the midgut proteins ofBombyx mori. Binding and competition assays revealed that GTase, MTase and VP4 played roles as viral attachment proteins. Far-Western blotting and LC-MS/MS analyses identified that heat shock protein 70 (BmHSP70), glutamate dehydrogenase (BmGDH), and angiotensin-converting enzyme (BmACE) in the midgut proteins as ligand candidates of the viral attachment proteins, and this was further verified by co-immunoprecipitation and fluorescence co-localization assays. Viral binding to the host midgutin vitrowas inhibited by pre-treatingB. morimidgut proteins with anti-BmHSP70, anti-BmGDH, anti-BmACE antibodies singly and in combination. Incubating DpCPV-1 virions with prokaryotically expressed BmHSP70, BmGDH, and BmACE also decreased viral attachment to the host midgut.In vivobioassays revealed that viral infection inHelicoverpa armigerawas partially neutralized by BmHSP70, BmGDH, and BmACE. Taking together, we concluded that HSP70, GDH, and ACE mediate DpCPV attachment and entry via binding to the viral attachment proteins, VP3 and VP4. The findings provide foundation for further understanding the entry mechanisms of cypoviruses.
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