期刊
INSECT BIOCHEMISTRY AND MOLECULAR BIOLOGY
卷 93, 期 -, 页码 19-26出版社
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.ibmb.2017.12.003
关键词
Planthopper; CRISPR/Cas9; Cinnabar; White; Hemiptera
资金
- National Science Found for Excellent Young Scholars [31522047]
- National Natural Science Foundation of China [31772158]
- Zhejiang Provincial Natural Science Found for Distinguished Young Scholars [LR16C140001]
- Fundamental Research Funds for the Central Universities [2015XZZX004-34]
The brown planthopper Nilaparvata lugens is one of the most destructive insect pests in Asia, demonstrating high fertility and causing huge crop losses by sucking sap of rice as well as transmitting viruses. However, functional genomic studies on N. lugens are seriously constrained by lack of genetic tools. Here, we employed two eye pigmentation genes to generate germ-line mutations in N. lugens using the CRISPR/Cas9 (clustered regularly interspaced palindromic repeats/CRISPR-associated) system. We showed that injection of single guide RNA of the cinnabar gene of N. lugens (Nl-cn) into pre-blastoderm eggs induced insertion and deletion (indels) in the founder generation (Go), which were heritably transmitted to the following G(1) generation, leading to bright red compound eyes and ocelli. Mutations of N. lugens white (Nl-w) generated a high mutant rate of up to 27.3%, resulting in mosaic eyes consisting of white and lightly pigmented ommatidia in both G(0) and G(1) individuals. The specificity of CRISPR/Cas9-mediated mutagenesis was further bolstered by PCR and RNA interference-based knockdown analysis. These results show that CRISPR/Cas9-mediated gene editing is achievable in a hemipteran insect, offering a valuable tool for the study of functional genomics and pest management in this planthopper species. (C) 2017 Elsevier Ltd. All rights reserved.
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