CRISPR/Cas9 mediated knockout of the abdominal-A homeotic gene in the global pest, diamondback moth (Plutella xylostella)

被引:99
作者
Huang, Yuping [1 ,2 ,3 ]
Chen, Yazhou [4 ]
Zeng, Baosheng [4 ]
Wang, Yajun [1 ,2 ,3 ]
James, Anthony A. [5 ,6 ]
Gurr, Geoff M. [1 ,2 ,3 ,7 ]
Yang, Guang [1 ,2 ,3 ]
Lin, Xijian [1 ,2 ,3 ]
Huang, Yongping [4 ]
You, Minsheng [1 ,2 ,3 ]
机构
[1] Fujian Agr & Forestry Univ, Inst Appl Ecol, Fuzhou 350002, Peoples R China
[2] Fujian Agr & Forestry Univ, Fujian Taiwan Joint Ctr Ecol Control Crop Pests, Fuzhou 350002, Peoples R China
[3] Minist Agr, Key Lab Integrated Pest Management Fujian Taiwan, Fuzhou 350002, Peoples R China
[4] Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Plant Physiol & Ecol, Key Lab Insect Dev & Evolutionary Biol, Shanghai 200032, Peoples R China
[5] Univ Calif Irvine, Dept Microbiol & Mol Genet, Irvine, CA 92697 USA
[6] Univ Calif Irvine, Dept Mol Biol & Biochem, Irvine, CA 92697 USA
[7] Charles Sturt Univ, Graham Ctr, Orange, NSW 2800, Australia
基金
中国国家自然科学基金;
关键词
Pest; Lepidoptera; Germline stability; Phylogenetic analysis; Gene mutation; BITHORAX-COMPLEX; ABD-A; BOMBYX-MORI; DROSOPHILA-MELANOGASTER; HUMAN-CELLS; GENOME; NUCLEASE; PROTEINS; REQUIRES; SYSTEM;
D O I
10.1016/j.ibmb.2016.06.004
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The diamondback moth, Plutella xylostella (L.), is a worldwide agricultural pest that has developed resistance to multiple classes of insecticides. Genetics-based approaches show promise as alternative pest management approaches but require functional studies to identify suitable gene targets. Here we use the CRISPR/Cas9 system to target a gene, abdominal-A, which has an important role in determining the identity and functionality of abdominal segments. We report that P. xylostella abdominal-A (Pxabd-A) has two structurally-similar splice isoforms (A and B) that differ only in the length of exon II, with 15 additional nucleotides in isoform A. Pxabd-A transcripts were detected in all developmental stages, and particularly in pupae and adults. CRISPR/Cas9-based mutagenesis of Pxabd-A exon I produced 91% chimeric mutants following injection of 448 eggs. Phenotypes with abnormal prolegs and malformed segments were visible in hatched larvae and unhatched embryos, and various defects were inherited by the next generation (G(1)). Genotyping of mutants demonstrated several mutations at the Pxabd-A genomic locus. The results indicate that a series of insertions and deletions were induced in the Pxabd-A locus, not only in Go survivors but also in G1 individuals, and this provides a foundation for genome editing. Our study demonstrates the utility of the CRISPR/Cas9 system for targeting genes in an agricultural pest and therefore provides a foundation the development of novel pest management tools. (C) 2016 Elsevier Ltd. All rights reserved.
引用
收藏
页码:98 / 106
页数:9
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