Genetic pest control strategies based on precise sex separation and only releasing sterile males can be accomplished by site-specific genome editing.In the current study,we showed that the mutation of single-allele Px...Genetic pest control strategies based on precise sex separation and only releasing sterile males can be accomplished by site-specific genome editing.In the current study,we showed that the mutation of single-allele Pxfl(2)d can significantly impair the normal mating behavior and testis development in male adults of the notorious cruciferous insect pest Plutella xylostella,in addition to its known functions in the ovarian development in female adults and egg hatching.Subsequent CRISPR/Cas9-based knock-in experiments revealed that site-specific integration of an exogenous green fluorescent protein(GFP)gene into autosomal Pxfl(2)d for labelling mutants could be achieved.However,this gene is not a suitable target for GFP insertion to establish a genetically stable knock-in strain because of the severe decline in reproductive capacity.We further screened for the W-chromosome-linked and Z-chromosome-linked regions to test the knock-in efficiency mediated by CRISPR/Cas9.The results verified that both types of chromosomes can be targeted for the site-specific insertion of exogenous sequences.We ultimately obtained a homozygous knock-in strain with the integration of both Cas9 and cyan fluorescent protein(CFP)expression cassettes on a Z-linked region in P.xylostella,which can also be used for early sex detection.By injecting the sgRNA targeting Pxfl(2)d alone into the eggs laid by female adults of the Z-Cas9-CFP strain,the gene editing efficiency reached 29.73%,confirming the success of expressing a functional Cas9 gene.Taken together,we demonstrated the feasibility of the knock-in of an exogenous gene to different genomic regions in P.xylostella,while the establishment of a heritable strain required the positioning of appropriate sites.This study provides an important working basis and technical support for further developing genetic strategies for insect pest control.展开更多
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 diferent classes of insecticides,making it ...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 diferent classes of insecticides,making it increasingly dificult 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 Ist to 3rd instar larvae,pupae,and adults,indicating that Pxyellow plays an essential role in the body pigmentation of P xlostella.Fitness analysis revealed no significant difference in the oviposition of adults,the hatchability of eggs,and the weight of pupac 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 genctcally based pest control techniques using Pxyellow as a screening marker.展开更多
Tea green leafhopper(TGL),Empoasca onukii,is of biological and economic interest.Despite numerous studies,the mechanisms underlying its adaptation and evolution remain enigmatic.Here,we use previously untapped genome ...Tea green leafhopper(TGL),Empoasca onukii,is of biological and economic interest.Despite numerous studies,the mechanisms underlying its adaptation and evolution remain enigmatic.Here,we use previously untapped genome and population genetics approaches to examine how the pest adapted to different environmental variables and thus has expanded geographically.We complete a chromosome-level assembly and annotation of the E.onukii genome,showing notable expansions of gene families associated with adaptation to chemoreception and detoxification.Genomic signals indicating balancing selection highlight metabolic pathways involved in adaptation to a wide range of tea varieties grown across ecologically diverse regions.Patterns of genetic variations among 54 E.onukii samples unveil the population structure and evolutionary history across different tea-growing regions in China.Our results demonstrate that the genomic changes in key pathways,including those linked to metabolism,circadian rhythms,and immune system functions,may underlie the successful spread and adaptation of E.onukii.This work highlights the genetic and molecular basis underlying the evolutionary success of a species with broad economic impacts,and provides insights into insect adaptation to host plants,which will ultimately facilitate more sustainable pest management.展开更多
基金supported by the National Natural Science Foundation of China(32172503 and 32260721)the Natural Science Foundation of Fujian Province,China(2023J01069)+2 种基金the State Key Laboratory of Ecological Pest Control for Fujian and Taiwan Crops,China(SKL2022001)the Innovation Fund of Fujan Agriculture and Forestry University,China(KFB23014A)the Undergraduate Training Program for Innovation and Entrepreneurship of Fujian Province,China(S202210389101).
文摘Genetic pest control strategies based on precise sex separation and only releasing sterile males can be accomplished by site-specific genome editing.In the current study,we showed that the mutation of single-allele Pxfl(2)d can significantly impair the normal mating behavior and testis development in male adults of the notorious cruciferous insect pest Plutella xylostella,in addition to its known functions in the ovarian development in female adults and egg hatching.Subsequent CRISPR/Cas9-based knock-in experiments revealed that site-specific integration of an exogenous green fluorescent protein(GFP)gene into autosomal Pxfl(2)d for labelling mutants could be achieved.However,this gene is not a suitable target for GFP insertion to establish a genetically stable knock-in strain because of the severe decline in reproductive capacity.We further screened for the W-chromosome-linked and Z-chromosome-linked regions to test the knock-in efficiency mediated by CRISPR/Cas9.The results verified that both types of chromosomes can be targeted for the site-specific insertion of exogenous sequences.We ultimately obtained a homozygous knock-in strain with the integration of both Cas9 and cyan fluorescent protein(CFP)expression cassettes on a Z-linked region in P.xylostella,which can also be used for early sex detection.By injecting the sgRNA targeting Pxfl(2)d alone into the eggs laid by female adults of the Z-Cas9-CFP strain,the gene editing efficiency reached 29.73%,confirming the success of expressing a functional Cas9 gene.Taken together,we demonstrated the feasibility of the knock-in of an exogenous gene to different genomic regions in P.xylostella,while the establishment of a heritable strain required the positioning of appropriate sites.This study provides an important working basis and technical support for further developing genetic strategies for insect pest control.
基金the National Natural Science Foundation of China(31972271)the Strait Postdoctoral Exchange Program of Fujian(2018B002)Fujian Science and Technology Major Program(2018NZ01010013).
文摘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 diferent classes of insecticides,making it increasingly dificult 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 Ist to 3rd instar larvae,pupae,and adults,indicating that Pxyellow plays an essential role in the body pigmentation of P xlostella.Fitness analysis revealed no significant difference in the oviposition of adults,the hatchability of eggs,and the weight of pupac 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 genctcally based pest control techniques using Pxyellow as a screening marker.
基金supported by the National Key R&D Program of China(Grant No.2019YFD1002100)the Natural Science Foundation of Fujian Province,China(Grant No.2020J01525)+1 种基金the Fujian Agriculture and Forestry University Construction Project for Technological Innovation and Service System of Tea Industry,China(Grant No.K1520005A03)the Key International Science and Technology cooperation Project of China(Grant No.2016YFE0102100).
文摘Tea green leafhopper(TGL),Empoasca onukii,is of biological and economic interest.Despite numerous studies,the mechanisms underlying its adaptation and evolution remain enigmatic.Here,we use previously untapped genome and population genetics approaches to examine how the pest adapted to different environmental variables and thus has expanded geographically.We complete a chromosome-level assembly and annotation of the E.onukii genome,showing notable expansions of gene families associated with adaptation to chemoreception and detoxification.Genomic signals indicating balancing selection highlight metabolic pathways involved in adaptation to a wide range of tea varieties grown across ecologically diverse regions.Patterns of genetic variations among 54 E.onukii samples unveil the population structure and evolutionary history across different tea-growing regions in China.Our results demonstrate that the genomic changes in key pathways,including those linked to metabolism,circadian rhythms,and immune system functions,may underlie the successful spread and adaptation of E.onukii.This work highlights the genetic and molecular basis underlying the evolutionary success of a species with broad economic impacts,and provides insights into insect adaptation to host plants,which will ultimately facilitate more sustainable pest management.
