caa a22 4500 445844728 CHVBK 20180317145355.0 cr unu---uuuuu 170323e20110101xx s 000 0 eng 10.1007/s10709-010-9484-6 doi (NATIONALLICENCE)springer-10.1007/s10709-010-9484-6 Development of transgenic strains for the biological control of the Mexican fruit fly, Anastrepha ludens [Elektronische Daten] [J. Meza, Xavier Nirmala, Grazyna Zimowska, C. Zepeda-Cisneros, Alfred Handler] The Mexican fruit fly, Anastrepha ludens, is a highly significant agricultural pest species that has been genetically transformed with a piggyBac-based transposon vector system using independent vector and transposase helper plasmids. Minimum estimated germ-line transformation frequencies were approximately 13-21% per fertile G0 individual, similar to previously reported frequencies using single vector-helper plasmids. Two vector constructs were tested with potential importance to transgenic strain development for mexfly biological control. The first allows post-integration stabilization of a transposon-vector by deletion of a terminal sequence necessary for mobilization. The complete pB[L1-EGFP-L2-DsRed-R1] vector was integrated into the Chiapas wild type strain with subsequent deletion of the L2-DsRed-R1 sub-vector carrying the piggyBac 3′ terminal sequence. Quality control tests for three of the stabilization vector lines (previous to stabilization) assessed viability at all life stages, fertility, adult flight ability, and adult male sexual competitiveness. All three transgenic lines were less fit compared to the wild strain by approximately 5-10% in most tests, however, there was no significant difference in sexual competitiveness which is the major prerequisite for optimal strain release. The second vector, pB[XL-EGFP, Asß2-tub-DsRed.T3], has the DsRed.T3 fluorescent protein reporter gene regulated by the A. suspensa Asß2-tubulin promoter, that resulted in testis and sperm-specific DsRed fluorescence in transgenic male mexflies. Fluorescent sperm bundles were unambiguously observed in the spermathecae of non-transgenic females mated to transgenic males. One transgenic line apparently had a male-specific Y-chromosome insertion, having potential use for sexing by fluorescent-embryo sorting. All transgenic lines expressed easily detectable and stable fluorescence in adults allowing their identification after trapping in the field. Springer Science + Business Media B.V., 2010 Anastrepha ludens nationallicence Tephritid fruit flies nationallicence Insect transgenesis nationallicence Vector stabilization nationallicence Sperm marker nationallicence Sterile insect technique nationallicence Meza J. Campaña Nacional Moscas de la Fruta, DGSV-SAGARPA, Central Poniente No. 14 Altos, C.P. 30700, Tapachula, Chiapas, México aut Nirmala Xavier U.S. Department of Agriculture, Agricultural Research Service, Center for Medical, Agricultural, and Veterinary Entomology, 1700 SW 23rd Drive, 32608, Gainesville, FL, USA aut Zimowska Grazyna U.S. Department of Agriculture, Agricultural Research Service, Center for Medical, Agricultural, and Veterinary Entomology, 1700 SW 23rd Drive, 32608, Gainesville, FL, USA aut Zepeda-Cisneros C. Campaña Nacional Moscas de la Fruta, DGSV-SAGARPA, Central Poniente No. 14 Altos, C.P. 30700, Tapachula, Chiapas, México aut Handler Alfred U.S. Department of Agriculture, Agricultural Research Service, Center for Medical, Agricultural, and Veterinary Entomology, 1700 SW 23rd Drive, 32608, Gainesville, FL, USA aut Genetica Springer Netherlands 139/1(2011-01-01), 53-62 0016-6707 139:1<53 2011 139 10709 https://doi.org/10.1007/s10709-010-9484-6 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10709-010-9484-6 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Meza J. Campaña Nacional Moscas de la Fruta, DGSV-SAGARPA, Central Poniente No. 14 Altos, C.P. 30700, Tapachula, Chiapas, México aut NATIONALLICENCE 700 1- Nirmala Xavier U.S. Department of Agriculture, Agricultural Research Service, Center for Medical, Agricultural, and Veterinary Entomology, 1700 SW 23rd Drive, 32608, Gainesville, FL, USA aut NATIONALLICENCE 700 1- Zimowska Grazyna U.S. Department of Agriculture, Agricultural Research Service, Center for Medical, Agricultural, and Veterinary Entomology, 1700 SW 23rd Drive, 32608, Gainesville, FL, USA aut NATIONALLICENCE 700 1- Zepeda-Cisneros C. Campaña Nacional Moscas de la Fruta, DGSV-SAGARPA, Central Poniente No. 14 Altos, C.P. 30700, Tapachula, Chiapas, México aut NATIONALLICENCE 700 1- Handler Alfred U.S. Department of Agriculture, Agricultural Research Service, Center for Medical, Agricultural, and Veterinary Entomology, 1700 SW 23rd Drive, 32608, Gainesville, FL, USA aut NATIONALLICENCE 773 0- Genetica Springer Netherlands 139/1(2011-01-01), 53-62 0016-6707 139:1<53 2011 139 10709 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer