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How functional genomics will impact fruit fly pest control: the example of the Mediterranean fruit fly, Ceratitis capitata.

Scolari F, Gomulski LM, Gabrieli P, Manni M, Savini G, Gasperi G, Malacrida AR - BMC Genet. (2014)

Bottom Line: Extensive transcriptome resources and the recently released genome sequence are making it possible to unravel several aspects of the medfly reproductive biology and behaviour, opening new opportunities for comparative genomics and barcoding for species identification.New genes, promotors and regulatory sequences are becoming available for the development/improvement of highly competitive sexing strains, for the monitoring of sterile males released in the field and for determining the mating status of wild females.The tools developed in this species have been transferred to other tephritids that are also the subject of SIT programmes.

View Article: PubMed Central - HTML - PubMed

ABSTRACT
The highly invasive agricultural insect pest Ceratitis capitata (Diptera: Tephritidae) is the most thoroughly studied tephritid fruit fly at the genetic and molecular levels. It has become a model for the analysis of fruit fly invasions and for the development of area-wide integrated pest management (AW-IPM) programmes based on the environmentally-friendly Sterile Insect Technique (SIT). Extensive transcriptome resources and the recently released genome sequence are making it possible to unravel several aspects of the medfly reproductive biology and behaviour, opening new opportunities for comparative genomics and barcoding for species identification. New genes, promotors and regulatory sequences are becoming available for the development/improvement of highly competitive sexing strains, for the monitoring of sterile males released in the field and for determining the mating status of wild females. The tools developed in this species have been transferred to other tephritids that are also the subject of SIT programmes.

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Transgenic sperm can be easily traced in the reproductive tract of laboratory wild-type females. Mechanically opened spermatheca isolated from a laboratory wild-type female mated with a transgenic male with green fluorescent sperm [32], three days after death (A). Spermathecal duct dissected from a laboratory wild-type female mated to a transgenic male with green fluorescent sperm [32] 24 hours after mating (B). Images were captured using an epifluorescence Zeiss Axioplan microscope at 400x magnification with the Zeiss filters set 13.
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Figure 3: Transgenic sperm can be easily traced in the reproductive tract of laboratory wild-type females. Mechanically opened spermatheca isolated from a laboratory wild-type female mated with a transgenic male with green fluorescent sperm [32], three days after death (A). Spermathecal duct dissected from a laboratory wild-type female mated to a transgenic male with green fluorescent sperm [32] 24 hours after mating (B). Images were captured using an epifluorescence Zeiss Axioplan microscope at 400x magnification with the Zeiss filters set 13.

Mentions: A major issue in the monitoring activities for evaluating SIT effectiveness is the difficulty in assessing the capacity of released sterile males to inseminate wild-type females [60-62]. The availability of the testes- and sperm-specific Ccβ2-tubulin gene has allowed the use of its promotor for fluorescent protein marking of the spermatozoa, and hence to detect females that have mated with released males [32]. Using this marking system, strains have been generated and evaluated for their ability to transfer green or red fluorescent sperm to the female spermathecae. It has been proven that these sperm remain viable and fluorescent for a long time within the spermathecae, also after female death [32] (Figure 3). The transgene previously inserted in one of these lines, namely 1260_F-3_m-1, was then efficiently modified by the use of the site-specific integration system from phage phiC31 [34]. Post-integrational excision of one of the piggyBac inverted terminal repeats resulted in stably integrated transgene insertions that, being inert to the piggyBac transposase, could not be remobilized. This allowed the development of an optimized strain for pest control that minimizes environmental concerns (stab_1260_F-3_m-1)[34]. Once integrated into the medfly GSS Vienna-8 strain, this sperm marking system may offer valuable alternatives to the currently used fluorescent powders [63] that are detected in trapped flies using UV light. Moreover, this sperm marking system can also be integrated into strains carrying diverse transgenes in tandem, for example with conditional embryonic lethality [33] and sexing systems [31].


How functional genomics will impact fruit fly pest control: the example of the Mediterranean fruit fly, Ceratitis capitata.

Scolari F, Gomulski LM, Gabrieli P, Manni M, Savini G, Gasperi G, Malacrida AR - BMC Genet. (2014)

Transgenic sperm can be easily traced in the reproductive tract of laboratory wild-type females. Mechanically opened spermatheca isolated from a laboratory wild-type female mated with a transgenic male with green fluorescent sperm [32], three days after death (A). Spermathecal duct dissected from a laboratory wild-type female mated to a transgenic male with green fluorescent sperm [32] 24 hours after mating (B). Images were captured using an epifluorescence Zeiss Axioplan microscope at 400x magnification with the Zeiss filters set 13.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4255779&req=5

Figure 3: Transgenic sperm can be easily traced in the reproductive tract of laboratory wild-type females. Mechanically opened spermatheca isolated from a laboratory wild-type female mated with a transgenic male with green fluorescent sperm [32], three days after death (A). Spermathecal duct dissected from a laboratory wild-type female mated to a transgenic male with green fluorescent sperm [32] 24 hours after mating (B). Images were captured using an epifluorescence Zeiss Axioplan microscope at 400x magnification with the Zeiss filters set 13.
Mentions: A major issue in the monitoring activities for evaluating SIT effectiveness is the difficulty in assessing the capacity of released sterile males to inseminate wild-type females [60-62]. The availability of the testes- and sperm-specific Ccβ2-tubulin gene has allowed the use of its promotor for fluorescent protein marking of the spermatozoa, and hence to detect females that have mated with released males [32]. Using this marking system, strains have been generated and evaluated for their ability to transfer green or red fluorescent sperm to the female spermathecae. It has been proven that these sperm remain viable and fluorescent for a long time within the spermathecae, also after female death [32] (Figure 3). The transgene previously inserted in one of these lines, namely 1260_F-3_m-1, was then efficiently modified by the use of the site-specific integration system from phage phiC31 [34]. Post-integrational excision of one of the piggyBac inverted terminal repeats resulted in stably integrated transgene insertions that, being inert to the piggyBac transposase, could not be remobilized. This allowed the development of an optimized strain for pest control that minimizes environmental concerns (stab_1260_F-3_m-1)[34]. Once integrated into the medfly GSS Vienna-8 strain, this sperm marking system may offer valuable alternatives to the currently used fluorescent powders [63] that are detected in trapped flies using UV light. Moreover, this sperm marking system can also be integrated into strains carrying diverse transgenes in tandem, for example with conditional embryonic lethality [33] and sexing systems [31].

Bottom Line: Extensive transcriptome resources and the recently released genome sequence are making it possible to unravel several aspects of the medfly reproductive biology and behaviour, opening new opportunities for comparative genomics and barcoding for species identification.New genes, promotors and regulatory sequences are becoming available for the development/improvement of highly competitive sexing strains, for the monitoring of sterile males released in the field and for determining the mating status of wild females.The tools developed in this species have been transferred to other tephritids that are also the subject of SIT programmes.

View Article: PubMed Central - HTML - PubMed

ABSTRACT
The highly invasive agricultural insect pest Ceratitis capitata (Diptera: Tephritidae) is the most thoroughly studied tephritid fruit fly at the genetic and molecular levels. It has become a model for the analysis of fruit fly invasions and for the development of area-wide integrated pest management (AW-IPM) programmes based on the environmentally-friendly Sterile Insect Technique (SIT). Extensive transcriptome resources and the recently released genome sequence are making it possible to unravel several aspects of the medfly reproductive biology and behaviour, opening new opportunities for comparative genomics and barcoding for species identification. New genes, promotors and regulatory sequences are becoming available for the development/improvement of highly competitive sexing strains, for the monitoring of sterile males released in the field and for determining the mating status of wild females. The tools developed in this species have been transferred to other tephritids that are also the subject of SIT programmes.

Show MeSH
Related in: MedlinePlus