Germline transformation of the spotted wing drosophilid, Drosophila suzukii, with a piggyBac transposon vector

Genetica ◽  
2013 ◽  
Vol 141 (4-6) ◽  
pp. 189-193 ◽  
Author(s):  
Marc F. Schetelig ◽  
Alfred M. Handler
Keyword(s):  
Drosophila Suzukii ◽  
Piggybac Transposon ◽  
Germline Transformation

scholarly journals Improvement on the genetic engineering of an invasive agricultural pest insect, the cherry vinegar fly, Drosophila suzukii

BMC Genetics ◽  
2020 ◽  
Vol 21 (S2) ◽  
Author(s):  
Hassan M. M. Ahmed ◽  
Fabienne Heese ◽  
Ernst A. Wimmer
Keyword(s):  
Genetic Engineering ◽  
Control Strategies ◽  
Landing Site ◽  
Drosophila Suzukii ◽  
French Alps ◽  
Germline Transformation ◽  
Φc31 Integrase ◽  
Transgenic Lines ◽  
Vinegar Fly ◽  
Driver Line

Abstract Background The invasive fly Drosophila suzukii has become an established fruit pest in Europe, the USA, and South America with no effective and safe pest management. Genetic engineering enables the development of transgene-based novel genetic control strategies against insect pests and disease vectors. This, however, requires the establishment of reliable germline transformation techniques. Previous studies have shown that D. suzukii is amenable to transgenesis using the transposon-based vectors piggyBac and Minos, site-specific recombination (lox/Cre), and CRISPR/Cas9 genome editing. Results We experienced differences in the usability of piggyBac-based germline transformation in different strains of D. suzukii: we obtained no transgenic lines in a US strain, a single rare transgenic line in an Italian strain, but observed a reliable transformation rate of 2.5 to 11% in a strain from the French Alps. This difference in efficiency was confirmed by comparative examination of these three strains. In addition, we used an attP landing site line to successfully established φC31-integrase-mediated plasmid integration at a rate of 10% and generated landing site lines with two attP sequences to effectively perform φC31-Recombinase Mediated Cassette Exchange (φC31-RMCE) with 11% efficiency. Moreover, we isolated and used the endogenous regulatory regions of Ds nanos to express φC31 integrase maternally to generate self-docking lines for φC31-RMCE. Besides, we isolated the promoter/enhancer of Ds serendipity α to drive the heterologous tetracycline-controlled transactivator (tTA) during early embryonic development and generated a testes-specific tTA driver line using the endogenous beta-2-tubulin (β2t) promoter/enhancer. Conclusion Our results provide evidence that the D. suzukii strain AM derived from the French Alps is more suitable for piggyBac germline transformation than other strains. We demonstrated the feasibility of using φC31-RMCE in the cherry vinegar fly and generated a set of lines that can be used for highly efficient integration of larger constructs. The φC31-based integration will facilitate modification and stabilization of previously generated transgenic lines that carry at least one attP site in the transgene construction. An early embryo-specific and a spermatogenesis-specific driver line were generated for future use of the binary expression system tet-off to engineer tissue- and stage-specific effector gene expression for genetic pest control strategies.

Germline transformation of the silkworm Bombyx mori L. using a piggyBac transposon-derived vector

10.1038/71978 ◽  
2000 ◽  
Vol 18 (1) ◽  
pp. 81-84 ◽  
Author(s):  
Toshiki Tamura ◽  
Chantal Thibert ◽  
Corinne Royer ◽  
Toshio Kanda ◽  
Abraham Eappen ◽  
...  
Keyword(s):  
Bombyx Mori ◽  
Piggybac Transposon ◽  
Germline Transformation ◽  
Bombyx Mori L ◽  
Silkworm Bombyx Mori

Effects of insecticides on adults and eggs of Drosophila suzukii (Diptera, Drosophilidae)

2017 ◽  
Vol 43 (2) ◽  
pp. 208 ◽  
Author(s):  
Daniele Cristine Hoffmann Schlesener ◽  
Jutiane Wollmann ◽  
Juliano De Bastos Pazini ◽  
Anderson Dionei Grützmacher ◽  
Flávio Roberto Mello Garcia
Keyword(s):  
North America ◽  
South America ◽  
Chemical Control ◽  
Stone Fruits ◽  
Drosophila Suzukii ◽  
Ovicidal Effect ◽  
Mere Presence ◽  
South Of Brazil ◽  
Different Cultures ◽  
First Time

Drosophila suzukii (Diptera, Drosophilidae) is an exotic species, endemic to Asia and currently a pest to small and stone fruits in several countries of North America and Europe. It was detected in 2013 for the first time in South America, in the south of Brazil. Unlike most drosophilids, this species deserves special attention, because the females are capable of oviposit inside healthy fruits, rendering their sale and export prohibited. Despite the confirmed existence of this species in different states of Brazil, this insect is yet been to be given the pest status. Nevertheless, the mere presence of this species is enough to cause concern to producers of small fruits and to justify further investigation for it’s control, especially chemical control for a possible change in status. Therefore, the goal of this work was to evaluate, in laboratory, mortality of D. suzukii adults and ovicidal effect when exposed to different insecticides registered for species of the Tephritidae and Agromyzidae families in different cultures. The insecticides deltamethrin, dimethoate, spinosad, fenitrothion, phosmet, malathion, methidathion, and zeta-cypermethrin resulted in mortality to 100 % of the subjects three days after the treatment (DAT). Regarding the effects over eggs, it was  established that the insecticides fenitrothion, malathion, and methidathion deemed 100 % of the eggs not viable, followed by phosmet and diflubenzuron, which also caused elevated reduction in the eclosion of larvae two DAT.

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