The sterile insect technique,which consists of the mass production and release of sterile insects to control populations of pests and disease vectors,has been effectively used for decades.An important component of ste...The sterile insect technique,which consists of the mass production and release of sterile insects to control populations of pests and disease vectors,has been effectively used for decades.An important component of sterile insect technique field applications is the availability of sex separation systems that reliably and economically eliminate females from mass-reared sterile insect populations destined for field release.Genetic sexing strains are important for the effectiveness and cost-efficiency of insect population control programs,including sterile insect technique.Classical approaches to generate genetic sexing strains,such as irradiation-induced chromosomal translocations,have yielded stable strains for species like the Mediterranean fruit fly,Ceratitis capitata.However,significant efforts are needed to establish genetic sexing strains using classical genetic methods,as large-scale random mutagenesis and screening are needed.We introduce here a neoclassical genetic approach,leveraging CRISPR-based gene-editing to target known genes to develop selectable genetic markers,followed by genetic rescue in a male-specific manner to speed up the development of genetic sexing strains and enhance their precision,stability,and adaptability.The integration of molecular tools,genetic markers like the white pupae and temperature-sensitive lethal,and strategies for maintaining genetic stability are discussed.We also review the challenges and opportunities in applying classical,transgenic,and neoclassical genetic approaches to improve genetic sexing strains for pest management.展开更多
The sterile insect technique(SIT)plays an important role in environmentally sustainable pest management.Its effectiveness hinges on specialized genetic tools called genetic sexing strains(GSSs),which enable the produc...The sterile insect technique(SIT)plays an important role in environmentally sustainable pest management.Its effectiveness hinges on specialized genetic tools called genetic sexing strains(GSSs),which enable the production and release of sterile male insects while excluding females.A collaborative initiative by the International Atomic Energy Agency(IAEA),designated as D44003“A generic approach for the development of genetic sexing strains for Sterile Insect Technique applications,”aimed to establish a universal framework for developing GSSs applicable to various pest species.This overview summarizes findings from 17 research studies,including laboratory models and field pests.The studies focused on enhancing genetic markers,genome editing,understanding sex determination,creating temperature-sensitive lethal systems,refining radiation protocols,and improving strain components.Collectively,these efforts demonstrate that a versatile,species-independent approach is both feasible and practical in real-world pest control efforts.This progress paves the way for the broader adoption of the SIT,promising a significant advance in integrated pest management.展开更多
The Bactrocera genus includes highly invasive fruit and vegetable pest species such as Bactrocera dorsalis,Bactrocera correcta,and Bactrocera oleae.The sterile insect technique(SIT)is a biological control method used ...The Bactrocera genus includes highly invasive fruit and vegetable pest species such as Bactrocera dorsalis,Bactrocera correcta,and Bactrocera oleae.The sterile insect technique(SIT)is a biological control method used to suppress populations of the invasive Bactrocera fruit flies by releasing sterilized male insects that compete with wild males for mates,reducing reproduction and eventually pest numbers.The effectiveness of the SIT against insect pests can be enhanced through male-only releases,achieved via genetic sexing strains(GSS)that enable early-stage sex separation.To overcome limitations faced when developing a new GSS through the classical genetic approach,a novel“neoclassical approach”has been proposed,focusing on the identification of genetic markers,the induction of desired phenotypes through genome editing,and the linkage of selectable markers to male sex.In this study,we evaluated the white pupae gene as a selectable marker for GSS development in 3 Bactrocera species.The white pupae orthologous genes have been identified,and,through CRISPR/Cas9 mutagenesis,the 3rd exon of the white pupae gene was knocked out resulting in white pupae lines in Bactrocera dorsalis,Bactrocera correcta,and Bactrocera oleae species.These results demonstrate the applicability of CRISPR/Cas9-mediated disruption of the conserved white pupae gene as a selectable marker in multiple Bactrocera species,supporting the development of genetic sexing systems for SIT-based pest management.展开更多
Methoprene （an analogue of juvenile hormone） application and feeding on a protein diet is known to enhance male melon fly, Bactrocera cucurbitae Coquillett （Diptera： Tephritidae）, mating success. In this study, w...Methoprene （an analogue of juvenile hormone） application and feeding on a protein diet is known to enhance male melon fly, Bactrocera cucurbitae Coquillett （Diptera： Tephritidae）, mating success. In this study, we investigated the effect of these treatments on male B. cucurbitae＇s ability to inhibit female remating. While 14-d-old females were fed on protein diet, 6-d-old males were exposed to one of the following treatments： （i） topical application of methoprene and fed on a protein diet; （ii） no methoprene but fed on a protein diet; （iii） methoprene and sugar-fed only; and （iv） sugar-fed, 14-d-old males acted as controls. Treatments had no effect on a male＇s ability to depress the female remating receptivity in comparison to the control. Females mated with protein-deprived males showed higher remating receptivity than females first mated with protein-fed males. Methoprene and protein diet interaction had a positive effect on male mating success during the first and second mating of females. Significantly more females first mated with sugar-fed males remated with protein-fed males and females first mated with methoprene treated and protein-fed males were more likely to remate with similarly treated males. Females mating latency （time to start mating） was significantly shorter with protein-fed males, and mating duration was significantly longer with protein-fed males compared with protein-deprived males. These results are discussed in the context of methoprene and/or dietary protein as prerelease treatment of sterile males in area-wide control of melon fly integrating the sterile insect technique （SIT）.展开更多
Methyl-eugenol （ME） （1,2-dimethoxy-4-（2-propenyl）benzene）, a natural phy- tochemical, did enhance male Bactrocera carambolae Drew ＆ Hancock （Diptera： Tephri- tidae） mating competitiveness 3 d after ingestion...Methyl-eugenol （ME） （1,2-dimethoxy-4-（2-propenyl）benzene）, a natural phy- tochemical, did enhance male Bactrocera carambolae Drew ＆ Hancock （Diptera： Tephri- tidae） mating competitiveness 3 d after ingestion. Enhanced male mating competitiveness can significantly increase the effectiveness of the sterile insect technique （SIT）. ME appli- cation to mass reared sterile flies by feeding is infeasible. ME application by aromatherapy however, would be a very practical way of ME application in fly emergence and release facilities. This approach was shown to enhance mating competitiveness ofB. carambolae 3 d posttreatment （DPT）. Despite this added benefit, every additional day of delaying re- lease will reduce sterile fly quality and will add cost to SIT application. The present study was planned to assess the effects of ME-aromatherapy on male B. carambolae mating competitiveness 1DPT and 2DPT. ME aromatherapy 1DPT or 2DPT did enhance mating competitiveness of B. carambolae males whereas ME feeding 1DPT and 2DPT did not. Male mating competitiveness was enhanced by the ME aromatherapy irrespective if they received 1DPT, 2DPT or 3DPT. ME aromatherapy, being a viable approach for its applica- tion, did enhance mating competitiveness of male B. carambolae 1 d posttreatrnent as ME feeding did 3 d after ingestion.展开更多
基金This study benefited from discussions at meetings for the Coordinated Research Project D44003 on the“Generic approach for the development of genetic sexing strains for SIT applications,”funded by the International Atomic Energy Agency(IAEA)Research work carried out at the Insect Pest Control Laboratory was supported by the Insect Pest Control Subprogramme of the Joint FAO/IAEA Centre of Nuclear Techniques in Food and Agriculture and the US State Department in the frame of the“Enhance Agency’s Capacity to Provide Support to Member States to Control Aedes Mosquitoes as Vectors of Human Pathogens,Particularly Zika Virus,Using Integrated Vector Management Approaches with a Sterile Insect Technique Component”project+2 种基金Additional funding was provided by the German-Israeli Project Cooperation of the German Research Foundation(SCHE 1833/7-1 and SCHE 1833/7-2 to MFS and PP)the European Union’s Horizon Europe Research and Innovation Program REACT(grant agreement number 101059523 to MFS and PP)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The sterile insect technique,which consists of the mass production and release of sterile insects to control populations of pests and disease vectors,has been effectively used for decades.An important component of sterile insect technique field applications is the availability of sex separation systems that reliably and economically eliminate females from mass-reared sterile insect populations destined for field release.Genetic sexing strains are important for the effectiveness and cost-efficiency of insect population control programs,including sterile insect technique.Classical approaches to generate genetic sexing strains,such as irradiation-induced chromosomal translocations,have yielded stable strains for species like the Mediterranean fruit fly,Ceratitis capitata.However,significant efforts are needed to establish genetic sexing strains using classical genetic methods,as large-scale random mutagenesis and screening are needed.We introduce here a neoclassical genetic approach,leveraging CRISPR-based gene-editing to target known genes to develop selectable genetic markers,followed by genetic rescue in a male-specific manner to speed up the development of genetic sexing strains and enhance their precision,stability,and adaptability.The integration of molecular tools,genetic markers like the white pupae and temperature-sensitive lethal,and strategies for maintaining genetic stability are discussed.We also review the challenges and opportunities in applying classical,transgenic,and neoclassical genetic approaches to improve genetic sexing strains for pest management.
基金This study benefited from discussions held during meetings of the Coordinated Research Project D44003,“Generic approach for the development of genetic sexing strains for SIT applications,”funded by the International Atomic Energy Agency(IAEA).
文摘The sterile insect technique(SIT)plays an important role in environmentally sustainable pest management.Its effectiveness hinges on specialized genetic tools called genetic sexing strains(GSSs),which enable the production and release of sterile male insects while excluding females.A collaborative initiative by the International Atomic Energy Agency(IAEA),designated as D44003“A generic approach for the development of genetic sexing strains for Sterile Insect Technique applications,”aimed to establish a universal framework for developing GSSs applicable to various pest species.This overview summarizes findings from 17 research studies,including laboratory models and field pests.The studies focused on enhancing genetic markers,genome editing,understanding sex determination,creating temperature-sensitive lethal systems,refining radiation protocols,and improving strain components.Collectively,these efforts demonstrate that a versatile,species-independent approach is both feasible and practical in real-world pest control efforts.This progress paves the way for the broader adoption of the SIT,promising a significant advance in integrated pest management.
基金supported by the Insect Pest Control Subprogramme of the Joint FAO/IAEA Centre of Nuclear Techniques in Food and Agriculture.Additional funding was provided by the European Union’s Horizon Europe Research and Innovation Program REACT(grant agreement number 101059523 to MFS and KDM)Further support was provided by the two postgraduate programs of the Department of Biochemistry and Biotechnology of the University of Thessaly(“Biotechnology-Quality Assessment in Nutrition and the Environment”and“Applications of Molecular Biology-Genetics-Diagnostic Biomarkers”)+2 种基金also by the project“Synthetic Biology:from omics technologies to genomic engineering(OMIC-ENGINE)”(MIS 5002636)which is implemented under the Action“Reinforcement of the Research and Innovation Infrastructure,”funded by the Operational Programme“Competitiveness,Entrepreneurship and Innovation”(NSRF 2014-2020)co-financed by Greece and the European Union(European Regional Development Fund)(to M-EG,and KM)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The Bactrocera genus includes highly invasive fruit and vegetable pest species such as Bactrocera dorsalis,Bactrocera correcta,and Bactrocera oleae.The sterile insect technique(SIT)is a biological control method used to suppress populations of the invasive Bactrocera fruit flies by releasing sterilized male insects that compete with wild males for mates,reducing reproduction and eventually pest numbers.The effectiveness of the SIT against insect pests can be enhanced through male-only releases,achieved via genetic sexing strains(GSS)that enable early-stage sex separation.To overcome limitations faced when developing a new GSS through the classical genetic approach,a novel“neoclassical approach”has been proposed,focusing on the identification of genetic markers,the induction of desired phenotypes through genome editing,and the linkage of selectable markers to male sex.In this study,we evaluated the white pupae gene as a selectable marker for GSS development in 3 Bactrocera species.The white pupae orthologous genes have been identified,and,through CRISPR/Cas9 mutagenesis,the 3rd exon of the white pupae gene was knocked out resulting in white pupae lines in Bactrocera dorsalis,Bactrocera correcta,and Bactrocera oleae species.These results demonstrate the applicability of CRISPR/Cas9-mediated disruption of the conserved white pupae gene as a selectable marker in multiple Bactrocera species,supporting the development of genetic sexing systems for SIT-based pest management.
文摘Methoprene （an analogue of juvenile hormone） application and feeding on a protein diet is known to enhance male melon fly, Bactrocera cucurbitae Coquillett （Diptera： Tephritidae）, mating success. In this study, we investigated the effect of these treatments on male B. cucurbitae＇s ability to inhibit female remating. While 14-d-old females were fed on protein diet, 6-d-old males were exposed to one of the following treatments： （i） topical application of methoprene and fed on a protein diet; （ii） no methoprene but fed on a protein diet; （iii） methoprene and sugar-fed only; and （iv） sugar-fed, 14-d-old males acted as controls. Treatments had no effect on a male＇s ability to depress the female remating receptivity in comparison to the control. Females mated with protein-deprived males showed higher remating receptivity than females first mated with protein-fed males. Methoprene and protein diet interaction had a positive effect on male mating success during the first and second mating of females. Significantly more females first mated with sugar-fed males remated with protein-fed males and females first mated with methoprene treated and protein-fed males were more likely to remate with similarly treated males. Females mating latency （time to start mating） was significantly shorter with protein-fed males, and mating duration was significantly longer with protein-fed males compared with protein-deprived males. These results are discussed in the context of methoprene and/or dietary protein as prerelease treatment of sterile males in area-wide control of melon fly integrating the sterile insect technique （SIT）.
文摘Methyl-eugenol （ME） （1,2-dimethoxy-4-（2-propenyl）benzene）, a natural phy- tochemical, did enhance male Bactrocera carambolae Drew ＆ Hancock （Diptera： Tephri- tidae） mating competitiveness 3 d after ingestion. Enhanced male mating competitiveness can significantly increase the effectiveness of the sterile insect technique （SIT）. ME appli- cation to mass reared sterile flies by feeding is infeasible. ME application by aromatherapy however, would be a very practical way of ME application in fly emergence and release facilities. This approach was shown to enhance mating competitiveness ofB. carambolae 3 d posttreatment （DPT）. Despite this added benefit, every additional day of delaying re- lease will reduce sterile fly quality and will add cost to SIT application. The present study was planned to assess the effects of ME-aromatherapy on male B. carambolae mating competitiveness 1DPT and 2DPT. ME aromatherapy 1DPT or 2DPT did enhance mating competitiveness of B. carambolae males whereas ME feeding 1DPT and 2DPT did not. Male mating competitiveness was enhanced by the ME aromatherapy irrespective if they received 1DPT, 2DPT or 3DPT. ME aromatherapy, being a viable approach for its applica- tion, did enhance mating competitiveness of male B. carambolae 1 d posttreatrnent as ME feeding did 3 d after ingestion.
