Abstract
The pine caterpillar moth, Dendrolimus punctatus, is a devastating forest pest. Genetic manipulation of this insect pest is limited due to the lack of genomic and functional genomic toolsets. Recently, CRISPR/Cas9 technology has been demonstrated to be a promising approach to modify the genome. To investigate gene functions during the embryogenesis, we introduced CRISPR/Cas9 system in D. punctatus to precisely and effectively manipulate gene expressions inmutant embryos. Compared to controls, knocking out of DpWnt-1, a gene well known for its role in the early body planning, led to high embryonic mortality. Among these mutants, 32.9% of the embryos and larvae showed an abnormal development. DpWnt-1 mutants predominantly exhibited abnormal posterior segments. In addition, multiple phenotypes were observed, including the loss of limbs and the head deformation, suggesting that DpWnt-1 signaling pathway is necessary for anterior segmentation and appendage development. Overall, our results demonstrate that CRISPR/Cas9 system is feasible and efficient in inducing mutations at a specific locus in D. punctatus. This study not only lays the foundation for characterizing gene functions in a non-model species, but also facilitates the future development of pest control alternatives for a major defoliator.
Highlights
The pine caterpillar moth Dendrolimus punctatus (Lepidoptera: Lasiocampidae) is one of the most destructive forest pests in China and Southeast Asia, where it attacks a variety of pine species and causes extensive forest damages (Billings, 1991; Zeng et al, 2010)
The expression of DpWnt-1 peaked at the very beginning, declined during the development, and reached the minimum level at the end of embryogenesis (Figure 2), suggesting that DpWnt-1 may play a vital role in D. punctatus during the early embryogenesis
Understanding the function of Wnt Family Member 1 (Wnt-1) is critical for exploring its potential role in pest management
Summary
The pine caterpillar moth Dendrolimus punctatus (Lepidoptera: Lasiocampidae) is one of the most destructive forest pests in China and Southeast Asia, where it attacks a variety of pine species and causes extensive forest damages (Billings, 1991; Zeng et al, 2010). The sterile insect technique (SIT) is an environmentally friendly insect control technology that relies on the release of large numbers of sterile males to mate with wild females to suppress pest population (Benedict and Robinson, 2003). The application of this method, is limited by the production of undesired females which need to be separated and eliminated. A modified SIT technique, the release of insects carrying a conditional dominant lethal gene (RIDL) can overcome this issue by inducing
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