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CRISPR/Cas9-based functional analysis of yellow gene in the diamondback moth,Plutella xylostella |
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| Authors: | Yajun Wang Yuping Huang Xuejiao Xu Zhaoxia Liu Jianyu Li Xue Zhan Guang Yang Minsheng You Shijun You |
| Institution: | 1. State Key Laboratory of Ecological Pest Control for Fujian-Taiwan Crops, Institute of Applied Ecology, Fujian Agriculture and Forestry University, Fuzhou, 350002 China
Joint International Research Laboratory of Ecological Pest Control, Ministry of Education, Fuzhou, 350002 China Key Laboratory of Integrated Pest Management for Fujian-Taiwan Crops, Ministry of Agriculture, Fuzhou, 350002 China These authors contribute equally to this work.;2. State Key Laboratory of Ecological Pest Control for Fujian-Taiwan Crops, Institute of Applied Ecology, Fujian Agriculture and Forestry University, Fuzhou, 350002 China Department of Physiology & Neurobiology, University of Connecticut, Storrs, CT, 06269 USA These authors contribute equally to this work.;3. State Key Laboratory of Ecological Pest Control for Fujian-Taiwan Crops, Institute of Applied Ecology, Fujian Agriculture and Forestry University, Fuzhou, 350002 China Joint International Research Laboratory of Ecological Pest Control, Ministry of Education, Fuzhou, 350002 China Key Laboratory of Integrated Pest Management for Fujian-Taiwan Crops, Ministry of Agriculture, Fuzhou, 350002 China;4. State Key Laboratory of Ecological Pest Control for Fujian-Taiwan Crops, Institute of Applied Ecology, Fujian Agriculture and Forestry University, Fuzhou, 350002 China Joint International Research Laboratory of Ecological Pest Control, Ministry of Education, Fuzhou, 350002 China Key Laboratory of Integrated Pest Management for Fujian-Taiwan Crops, Ministry of Agriculture, Fuzhou, 350002 China Fujian Key Laboratory for Monitoring and Integrated Management of Crop Pests, Institute of Plant Protection, Fujian Academy of Agricultural Sciences, Fuzhou, 350013 China;5. State Key Laboratory of Ecological Pest Control for Fujian-Taiwan Crops, Institute of Applied Ecology, Fujian Agriculture and Forestry University, Fuzhou, 350002 China |
| Abstract: | The diamondback moth, Plutella xylostella (L.), is an economically important pest of cruciferous crops worldwide. This pest is notorious for rapid evolution of the resistance to different classes of insecticides, making it increasingly difficult to control. Genetics-based control approaches, through manipulation of target genes, have been reported as promising supplements or alternatives to traditional methods of pest management. Here we identified a gene of pigmentation (yellow) in P. xylostella, Pxyellow, which encodes 1674 bp complementary DNA sequence with four exons and three introns. Using the clustered regularly interspersed palindromic repeats (CRISPR)/CRISPR-associated protein 9 system, we knocked out Pxyellow, targeting two sites in Exon III, to generate 272 chimeric mutants (57% of the CRISPR-treated individuals) with color-changed phenotypes of the 1st to 3rd instar larvae, pupae, and adults, indicating that Pxyellow plays an essential role in the body pigmentation of P. xylostella. Fitness analysis revealed no significant difference in the oviposition of adults, the hatchability of eggs, and the weight of pupae between homozygous mutants and wildtypes, suggesting that Pxyellow is not directly involved in regulation of growth, development, or reproduction. This work advances our understanding of the genetic and insect science molecular basis for body pigmentation of P. xylostella, and opens a wide avenue for development of the genetically based pest control techniques using Pxyellow as a screening marker. |
| Keywords: | CRISPR/Cas9 diamondback moth genetically based control novel marker yellow gene |