Anastrepha fraterculus is a cryptic species complex with at least eight morphotypes distributed across the Americas.Among them,A.fraterculus sp.1,present in Argentina,is a major pest impacting fresh fruit production.I...Anastrepha fraterculus is a cryptic species complex with at least eight morphotypes distributed across the Americas.Among them,A.fraterculus sp.1,present in Argentina,is a major pest impacting fresh fruit production.Integrated pest management strategies,including chemical control and trapping,are currently employed to mitigate its effects.Genetic sexing strains of A.fraterculus sp.1 are being evaluated for use in sterile insect technique programs.To support traditional and emerging control methods,this study aimed to enhance the genomic understanding of this morphotype.Individual female and male samples were sequenced using long-and short-read technologies.The female genome(760 Mb)was de novo assembled into 58 scaffolds and the male genome(750 Mb)into 68 scaffolds,with BUSCO completeness scores of 98.8%and 98.7%,respectively.Synteny analysis revealed complete scaffolds of the five autosomes and enabled near-complete reconstruction of the X and Y chromosomes.Gene prediction identified 17751 and 16535 protein-coding genes(for female and male genomes,respectively),with repetitive regions representing 46%of both genomes.Additionally,the mitochondrial genome was fully assembled and annotated.This comprehensive genomic resource reveals candidate genes for functional studies,including gene editing and RNA interference,as successfully applied in related tephritid species.These findings lay the foundation for innovative,complementary biocontrol tools against A.fraterculus.展开更多
The sterile insect technique(SIT)is a highly effective biologically-based method for the suppression of many insect pest populations.SIT efficacy could be improved by methods of male sterilization that avoid the use o...The sterile insect technique(SIT)is a highly effective biologically-based method for the suppression of many insect pest populations.SIT efficacy could be improved by methods of male sterilization that avoid the use of irradiation that can result in diminished fitness and mating competitiveness.Alternative sterilization methods include conditional disruption of genes for male fertility,or using their sperm-specific promoters to drive the expression of genes for lethal effectors.Testing has begun for the testis-specificβ2-tubulin gene,though additional male fertility genes are required for redundancy or replacement,and for species where theβ2-tubulin isoform does not exist or is not testis-specific.Here we had the goal of identifying and characterizing the sequence and transcriptional expression of two genes in the caribfly,Anastrepha suspensa,that are cognates of D.melanogaster spermatocyte-specific male fertility genes.In Drosophila,wampa encodes a coiled-coil dynein subunit required for axonemal assembly essential to microtubule-based sperm motility,while Prosα6T is a proteasome subunit gene required for spermatid individualization and nuclear maturation.In A.suspensa a cognate to wampa exhibited testis-specific transcript expression,which was minimal in both male and female body tissue.A Prosα6T cognate was not apparent in A.suspensa,but its constitutive isoform,Prosα6,expresses in male testes,but also in male and female body tissue.Thus,for A.suspensa,wampa remains a strong candidate gene for male sterility strategies for SIT including a direct target for gene-editing knockout and use of its promoter for testisspecific toxicity or cell death in conditional expression systems.展开更多
Bactrocera zonata is a highly invasive agricultural pest that causes extensive damage to fruit crops.The Sterile Insect Technique(SIT),a species-specific and environmentally friendly pest control method,significantly ...Bactrocera zonata is a highly invasive agricultural pest that causes extensive damage to fruit crops.The Sterile Insect Technique(SIT),a species-specific and environmentally friendly pest control method,significantly benefits from the availability of Genetic Sexing Strains(GSSs)that enable efficient mass production of males for sterile release.However,no GSS currently exists for B.zonata limiting SIT applications targeting this important invasive pest.Here,we report two key advancements toward GSS development in this species.First,we present a high-quality,chromosome-level genome assembly from male B.zonata,identifying two scaffolds derived from the Y chromosome,which represent potential targets for future male-specific genetic engineering.Second,we demonstrate the feasibility of CRISPR/Cas9 genome editing in B.zonata by generating stable,homozygous white-eye mutants through targeted disruption of the conserved white-eye gene.This visible,recessive phenotype serves as a proof-of-concept for developing selectable markers in this species.Together,these results provide foundational genomic and genetic tools to support the development of GSSs in B.zonata,advancing the potential for sustainable,genetics-based pest control strategies.展开更多
基金supported by the grant PICT 20210491(Agencia Nacional de Promoción Científica y Técnica)PDI087(Instituto Nacional de Tecnología Agropecuaria)the International Atomic Energy Agency Research Contract no.23402 as part of the Coordinated Research Project“Generic approach for the development of genetic sexing strains for SIT applications.Publication costs for this study were provided by the International Atomic Energy Agency as part of the Coordinated Research Project”“Generic approach for the development of genetic sexing strains for SIT applications”。
文摘Anastrepha fraterculus is a cryptic species complex with at least eight morphotypes distributed across the Americas.Among them,A.fraterculus sp.1,present in Argentina,is a major pest impacting fresh fruit production.Integrated pest management strategies,including chemical control and trapping,are currently employed to mitigate its effects.Genetic sexing strains of A.fraterculus sp.1 are being evaluated for use in sterile insect technique programs.To support traditional and emerging control methods,this study aimed to enhance the genomic understanding of this morphotype.Individual female and male samples were sequenced using long-and short-read technologies.The female genome(760 Mb)was de novo assembled into 58 scaffolds and the male genome(750 Mb)into 68 scaffolds,with BUSCO completeness scores of 98.8%and 98.7%,respectively.Synteny analysis revealed complete scaffolds of the five autosomes and enabled near-complete reconstruction of the X and Y chromosomes.Gene prediction identified 17751 and 16535 protein-coding genes(for female and male genomes,respectively),with repetitive regions representing 46%of both genomes.Additionally,the mitochondrial genome was fully assembled and annotated.This comprehensive genomic resource reveals candidate genes for functional studies,including gene editing and RNA interference,as successfully applied in related tephritid species.These findings lay the foundation for innovative,complementary biocontrol tools against A.fraterculus.
基金supported by the Biotechnology Risk Assessment Research Grants Program,grant no.2020-33522-32271,from the USDA National Institute of Food and Agriculture.
文摘The sterile insect technique(SIT)is a highly effective biologically-based method for the suppression of many insect pest populations.SIT efficacy could be improved by methods of male sterilization that avoid the use of irradiation that can result in diminished fitness and mating competitiveness.Alternative sterilization methods include conditional disruption of genes for male fertility,or using their sperm-specific promoters to drive the expression of genes for lethal effectors.Testing has begun for the testis-specificβ2-tubulin gene,though additional male fertility genes are required for redundancy or replacement,and for species where theβ2-tubulin isoform does not exist or is not testis-specific.Here we had the goal of identifying and characterizing the sequence and transcriptional expression of two genes in the caribfly,Anastrepha suspensa,that are cognates of D.melanogaster spermatocyte-specific male fertility genes.In Drosophila,wampa encodes a coiled-coil dynein subunit required for axonemal assembly essential to microtubule-based sperm motility,while Prosα6T is a proteasome subunit gene required for spermatid individualization and nuclear maturation.In A.suspensa a cognate to wampa exhibited testis-specific transcript expression,which was minimal in both male and female body tissue.A Prosα6T cognate was not apparent in A.suspensa,but its constitutive isoform,Prosα6,expresses in male testes,but also in male and female body tissue.Thus,for A.suspensa,wampa remains a strong candidate gene for male sterility strategies for SIT including a direct target for gene-editing knockout and use of its promoter for testisspecific toxicity or cell death in conditional expression systems.
基金Funding was provided by the European Union’s Horizon Europe Research and Innovation Program(REACTgrant agreement number 101059523 to PAP,GP and KM)the US-Israel Binational Agricultural Research and Development Fund(BARD-grant agreement number IS-5590-23 to PAP,GP,AH)+3 种基金the Hebrew University of Jerusalem and Zelman Cowen Academic Initiatives(ZCAI Joint Project 2021 number 0456 to PAP and SWB)Initial support was generously provided in the form of an International Fellowship to FK from the Research Fund for International Cooperation,Robert H.Smith Faculty of Agriculture,Food and Environment,HUJI and HUJI startup funds to PAP.Genome sequencing was financially supported by the Insect Pest Control Subprogramme of the Joint FAO/IAEA Centre of Nuclear Techniques in Food and Agriculture,the German Research Foundation through the Middle East Cooperation project 491548882(PAP)the Canada Foundation for Innovation grants 40104 and 35444(to Ioannis Ragousis)Publication cost for this study was provided by the International Atomic Energy Agency as part of the Coordinated Research Project“Generic approach for the development of genetic sexing strains for SIT applications”。
文摘Bactrocera zonata is a highly invasive agricultural pest that causes extensive damage to fruit crops.The Sterile Insect Technique(SIT),a species-specific and environmentally friendly pest control method,significantly benefits from the availability of Genetic Sexing Strains(GSSs)that enable efficient mass production of males for sterile release.However,no GSS currently exists for B.zonata limiting SIT applications targeting this important invasive pest.Here,we report two key advancements toward GSS development in this species.First,we present a high-quality,chromosome-level genome assembly from male B.zonata,identifying two scaffolds derived from the Y chromosome,which represent potential targets for future male-specific genetic engineering.Second,we demonstrate the feasibility of CRISPR/Cas9 genome editing in B.zonata by generating stable,homozygous white-eye mutants through targeted disruption of the conserved white-eye gene.This visible,recessive phenotype serves as a proof-of-concept for developing selectable markers in this species.Together,these results provide foundational genomic and genetic tools to support the development of GSSs in B.zonata,advancing the potential for sustainable,genetics-based pest control strategies.
