Abstract
Field population surveillance of a targeted insect pest species is critical in evaluating management programs such as the sterile insect technique. Fluorescent powder dyes currently used to distinguish released tephritids from the field population are not optimal in terms of reliability and human health issues. Genetically transformed tephritid species present the possibility of using fluorescent transgenes for marking. Here we studied the stability of DsRed fluorescence in transgenic flies maintained in aqueous torula yeast borax and propylene glycol. DsRed was stable in both solutions for three weeks by visual microscopic observations and could be used to unambiguously distinguish them from non-fluorescent wild type flies. To compensate for any potential ambiguity in visual identification a diagnostic PCR was developed that could specifically amplify the exotic heterologous marker gene. Therefore, the use of sterile transgenic insect strains carrying stably integrated fluorescent protein marker genes in biologically-based control programs could greatly improve released fly identification in pest management programs.
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References
Anwar M, Chambers DL, Ohinata K, Kobayashi RM (1971) Radiation-sterilization of the Mediterranean fruit fly (Diptera: Tephritidae): comparison of spermatogenesis in flies treated as pupae or adults. Ann Entomol Soc Am 64:627–633
Bevis BJ, Glick BS (2002) Rapidly maturing variants of the Discosoma red fluorescent protein (DsRed). Nat Biotechnol 20:83–87
Dominiak BC, Schinag L, Nicol H (2000) Impact of fluorescent marker dyes on emergence of sterile Queensland fruit fly Bactrocera tryoni (Froggatt) (Diptera: Tephritidae). Gen Appl Entomol 29:45–47
Dominiak BC, Sundraralingham S, Jessup AJ, Barchia IM (2002) Pupal weight as a key parameter for the quality of mass produced adult Queensland Fruit Fly Bactrocera tryoni (Froggatt) (Diptera: Tephritidae) in 1997/98. Gen Appl Entomol 31:17–24
Enkerlin W (2007) Guidance for packing, shipping, holding and release of sterile flies in area-wide fruit fly control programmes. Food and Agriculture Organization of the United Nations, Rome
Florida Fruit Fly Detection Manual (2004) Revision 6. Florida Department of Agriculture and Consumer Services and United States Department of Agriculture. Cooperative Florida Fruit Fly Detection Program
Hagler JR, Jackson CG (2001) Methods for marking insects: current techniques and future prospects. Annu Rev Entomol 46:511–543
Handler AM (2002) Prospects for using genetic transformation for improved SIT and new biocontrol methods. Genetica 116:137–149
Handler AM, Harrell RA (2001) Polyubiquitin-regulated DsRed marker for transgenic insects. Biotechniques 31:820–828
Heath RR, Epsky ND, Landolt PJ, Sivinski J (1993) Development of attractants for monitoring Caribbean fruit flies (Diptera: Tephritideae). Fla Entomol 76:233–244
Horn C, Wimmer EA (2000) A versatile vector set for animal transgenesis. Dev Genes Evol 210:630–637
Horn C, Schmid BG, Pogoda FS, Wimmer EA (2002) Fluorescent transformation markers for insect transgenesis. Insect Biochem Mol Biol 32:1221–1235
IAEA (2003) Trapping guidelines for area-wide fruit fly programmes. IAEA/FAO, TG/FFP. IAEA, Vienna
Klassen W, Curtis C (2005) History of the sterile insect technique. In: Dyck V, Hendrichs J, Robinson AS (eds) Sterile insect technique. Springer Netherlands, Dordrecht, pp 3–36
Knipling EF (1955) Possibilities of insect control or eradication through the use of sexually sterile males. J Econ Entomol 48:459–462
Lee HS, Simon JA, Lis JT (1988) Structure and expression of ubiquitin genes of Drosophila melanogaster. Mol Cell Biol 8:4727–4735
McInnis DO (1993) Size differences between normal and irradiated sperm heads in mated female Mediterranean fruit flies (Diptera: Tephniridae). Ann Entomol Soc Am 86:305–308
McInnis DO, Tam S, Grace C, Miyashita D (1994) Population suppression and sterility rates induced by variable sex ratio, sterile insect releases of Ceratitis capitata (Diptera: Tephritidae) in Hawaii. Ann Entomol Soc Am 87:231–240
Meza JS, Nirmala X, Zimowska GJ, Zepeda-Cisneros CS, Handler AM (2011) Development of transgenic strains for the biological control of the Mexican fruit fly, Anastrepha ludens. Genetica 139:53–62
Nirmala X, Zimowska GJ, Handler AM (2009) Characterization of the proteasome beta 2 subunit gene and its mutant allele in the tephritid fruit fly pest, Anastrepha suspensa. Insect Mol Biol 18:333–340
Parker A (2005) Mass-rearing for sterile insect release. In: Dyck V, Hendrichs J, Robinson AS (eds) Sterile insect technique. Springer Netherlands, Dordrecht, pp 209–232
Peloquin JJ, Miller TA (2000) Detection of enhanced green fluorescent protein DNA in pink bollworm through polymerase chain reaction. J Cotton Sci 4:28–33
Robinson AS, Franz G, Fisher K (1999) Genetic sexing strains in the medfly, Ceratitis capitata: development, mass rearing and field applications. Trends Entomol 2:81–104
Robinson AS, Franz G, Atkinson PW (2004) Insect transgenesis and its potential role in agriculture and human health. Insect Biochem Mol Biol 34:113–120
Schetelig MF, Caceres C, Zacharopoulou A, Franz G, Wimmer EA (2009) Conditional embryonic lethality to improve the sterile insect technique in Ceratitis capitata (Diptera: Tephritidae). BMC Biol 7:4
Scolari F, Schetelig MF, Bertin S, Malacrida AR, Gasperi G, Wimmer EA (2008) Fluorescent sperm marking to improve the fight against the pest insect Ceratitis capitata (Wiedemann; Diptera: Tephritidae). New Biotechnol 25:76–84
Sivinski J, Pereira R (2005) Do wing markings in fruit flies (Diptera: Tephritidae) have sexual significance? Fla Entomol 88:321–324
Thomas DB (2003) Nontarget insects captured in fruit fly (Diptera: Tephritidae) surveillance traps. J Econ Entomol 96:1732–1737
Thomas DB, Holler TC, Heath RR, Salinas EJ, Moses AL (2001) Trap-lure combinations for surveillance of Anastrepha fruit flies (Diptera: Tephritidae). Fla Entomol 84:344–351
Weldon CW (2005) Marking Queensland fruit fly, Bactrocera tryoni (Froggatt) (Diptera: Tephritidae) with fluorescent pigments: pupal emergence, adult mortality, and visibility and persistence of marks. Gen Appl Entomol 34:7–13
Zimowska GJ, Nirmala X, Handler AM (2009) The beta 2-tubulin gene from three tephritid fruit fly species and use of its promoter for sperm marking. Insect Biochem Mol Biol 39:508–515
Acknowledgments
The authors thank Drs. John Sivinski and Kenneth Bloem for critical comments on the manuscript and Jennifer Mestas and Alice Miano for technical assistance. Funding from the USDA-NIFA-Agriculture and Food Research Initiative is gratefully acknowledged.
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Nirmala, X., Olson, S.R., Holler, T.C. et al. A DsRed fluorescent protein marker under polyubiquitin promoter regulation allows visual and amplified gene detection of transgenic Caribbean fruit flies in field traps. BioControl 56, 333–340 (2011). https://doi.org/10.1007/s10526-011-9347-9
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DOI: https://doi.org/10.1007/s10526-011-9347-9