SeBLOS2 knockout via CRISPR/Cas9 leads to the loss of larval integument coloration in Spodoptera exigua (Lepidoptera: Noctuidae)

文献类型： 外文期刊

第一作者： Zhao, Jing

作者： Zhao, Jing;Jiang, Yiping;Tan, Yongan;Xiao, Liubin;Zhao, Jing;Hoffmann, Ary

作者机构：

关键词： CRISPR/Cas9; integument coloration; SeBLOS2; Spodoptera exigua

期刊名称：ARCHIVES OF INSECT BIOCHEMISTRY AND PHYSIOLOGY （ 影响因子：2.2； 五年影响因子：2.1 ）

ISSN： 0739-4462

年卷期： 2023 年

页码：

收录情况： SCI

摘要： CRISPR/Cas9 technology is a precise and powerful tool for functionally exploring insect genes. The present study tested CRISPR/Cas9 as a way of undertaking effective gene mutagenesis in an important agricultural pest, the beet armyworm Spodoptera exigua. Based on a S. exigua transcriptome database, the entire complementary DNA sequence of SeBLOS2 encoding 140 amino acid residues was cloned. The gene was highly expressed in late larval stages (L3-L5). Using the CRISPR/Cas9 method, SeBLOS2 was knocked out by altering two sites in the coding region. This resulted in 70%-74% of the G0 generation (L4-L5) larvae displaying mosaic translucent integument. Four different mutations occurred at SeBLOS2-specific target sites, as demonstrated by further polymerase chain reaction-based genotypic analysis. Homozygote mutant L3 larvae were obtained in the G1 generation, with complete loss of white stripes and spots on their larval integument. These results demonstrate a crucial role of SeBLOS2 in integument pigmentation and suggest that the gene can act as a suitable nonlethal marker for functional research on genes in S. exigua and other Lepidopteran pests.

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