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
作者： Zhao, Jing 1 ; Jiang, Yiping 1 ; Hoffmann, Ary 3 ; Tan, Yongan 1 ; Xiao, Liubin 1 ;
作者机构： 1.Jiangsu Acad Agr Sci, Inst Plant Protect, Nanjing, Peoples R China
2.Jiangsu Acad Agr Sci, Key Lab Food Qual, Safety Jiangsu Prov, State Key Lab Breeding Base, Nanjing, Peoples R China
3.Univ Melbourne, Inst Bio21, Fac Sci, Sch Biosci,Pest & Environm Adaptat Res Grp, Melbourne, Vic, Australia
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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