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Insect Biochem Mol Biol

Title:		CRISPR/Cas9 mediated gene knockout reveals a more important role of PBP1 than PBP2 in the perception of female sex pheromone components in Spodoptera litura
Author(s):		Zhu GH; Zheng MY; Sun JB; Khuhro SA; Yan Q; Huang Y; Syed Z; Dong SL;
Address:		"Education Ministry Key Laboratory of Integrated Management of Crop Disease and Pests, College of Plant Protection, Nanjing Agricultural University, Nanjing, 210095, China; Department of Entomology, University of Kentucky, Lexington, KY, 40546, USA. Education Ministry Key Laboratory of Integrated Management of Crop Disease and Pests, College of Plant Protection, Nanjing Agricultural University, Nanjing, 210095, China. Key Laboratory of Insect Developmental and Evolutionary Biology, Institute of Plant Physiology and Ecology, Chinese Academy of Sciences, Shanghai Institutes for Biological Sciences, Shanghai, 200032, China. Department of Entomology, University of Kentucky, Lexington, KY, 40546, USA. Education Ministry Key Laboratory of Integrated Management of Crop Disease and Pests, College of Plant Protection, Nanjing Agricultural University, Nanjing, 210095, China. Electronic address: sldong@njau.edu.cn"
Journal Title:		Insect Biochem Mol Biol
Year:		2019
Volume:		20190924
Issue:
Page Number:		103244 -
DOI:		10.1016/j.ibmb.2019.103244
ISSN/ISBN:		1879-0240 (Electronic) 0965-1748 (Linking)
Abstract:		"Three different pheromone binding proteins (PBPs) can typically be found in the sensilla lymph of noctuid moth antennae, but their relative contributions in perception of the sex pheromone is rarely verified in vivo. Previously, we demonstrated that SlitPBP3 plays a minor role in the sex pheromone detection in Spodoptera litura using the CRISPR/Cas9 system. In the present study, the roles of two other SlitPBPs (SlitPBP1 and SlitPBP2) are further verified using the same system. First, by co-injection of Cas9 mRNA/sgRNA into newly laid eggs, a high rate of target mutagenesis was induced, 51.5% for SlitPBP1 and 46.8% for SlitPBP2 as determined by restriction enzyme assay. Then, the homozygous SlitPBP1 and SlitPBP2 knockout lines were obtained by cross-breeding. Finally, using homozygous knockout male moths, we performed electrophysiological (EAG recording) and behavioral analyses. Results showed that knockout of either SlitPBP1 or SlitPBP2 in males decreased EAG response to each of the 3 sex pheromone components (Z9,E11-14:Ac, Z9,E12-14:Ac and Z9-14:Ac) by 53%, 60% and 63% (for SlitPBP1 knockout) and 40%, 43% and 46% (for SlitPBP2 knockout), respectively. These decreases in EAG responses were similar among 3 pheromone components, but were more pronounced in SlitPBP1 knockout males than in SlitPBP2 knockout males. Consistently, behavioral assays with the major component (Z9,E11-14:Ac) showed that SlitPBP1 knockout males responded in much lower percentages than SlitPBP2 knockout males in terms of orientation to the pheromone, along with reduction in close range behaviors such as hairpencil display and mating attempt. Taken together, this study provides direct functional evidence for the roles of SlitPBP1 and SlitPBP2, as well as their relative importance (SlitPBP1?ª+>?ª+SlitPBP2) in the sex pheromone perception. This information is valuable in understanding mechanisms of sex pheromone perception and may facilitate the development of PBP-targeted pest control techniques"
Keywords:		*Animal Communication Animals Arthropod Antennae/*physiology Base Sequence CRISPR-Cas Systems Carrier Proteins/*physiology Female Insect Proteins/*physiology Male Mutation *Olfactory Perception Sex Attractants Spodoptera/*physiology CRISPR/Cas9 Genome edi;
Notes:		"MedlineZhu, Guan-Heng Zheng, Mei-Yan Sun, Jia-Bin Khuhro, Sajjad Ali Yan, Qi Huang, Yongping Syed, Zainulabeuddin Dong, Shuang-Lin eng Research Support, Non-U.S. Gov't England 2019/09/29 Insect Biochem Mol Biol. 2019 Dec; 115:103244. doi: 10.1016/j.ibmb.2019.103244. Epub 2019 Sep 24"