Campana Nacional Moscas de la Fruta

Dolores Hidalgo Cuna de la Independencia Nacional, Mexico

Campana Nacional Moscas de la Fruta

Dolores Hidalgo Cuna de la Independencia Nacional, Mexico
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Ibanez-Palacios J.,Campana Nacional Moscas de la Fruta | Garcia-Velazquez A.,Colegio de Mexico | Zepeda-Cisneros C.S.,Campana Nacional Moscas de la Fruta | Corona-Torres T.,Colegio de Mexico
Agrociencia | Year: 2010

The genus Anastrepha Schiner, endemic to tropical and subtropical América, includes the species Anastrepha ludens, A. obliqua, A. striata and A. serpentina, which are polyphagous and parasite a wide range of plants. These species were studied because of their broad geographic distribution and because they parasite different fruit species. Larvae in their third stage reared in the laboratory of the Programa Moscafrut in Metapa de Domínguez, Chiapas, México, were used. The cerebral ganglia were dissected in an aqueous solution of 1 % (w/v) sodium citrate in which they were kept for 20 min, then transferred to methanol:acetic acid (3:1 (v/v) and macerated in 60 % acetic acid. One drop of this tissue suspension was placed on a slide on a hot plate (45-50 °C) and observed with a microscope (Zeiss Axioskop 40) with phase contrast, and the cells with disperse chromosomes were selected. These were photographed with an AxioCam camera and Axiovision 4.0 software, and measurements were taken with Image Tool 3.0. Twenty cells/sex/species were analyzed. The centromere was observed without pretreatment. Karyotypes were determined in each species: A. ludens {square with filled S-E corner}, 2n=12, Xg Xg+VII telo, n. f. 12, {black star}, 2n=12, Xg Yp+VII Telo n. f. 12; A. obliqua {square with filled+?VII Telo n. f. 12; A. striata {square with filled S-E corner}, 2n=12, Xg Xg+IIII Meta?IIIII Telo n. f. 16, {black star}, 2n=12, Xg Yp+IIII Meta?IIII Telo n. f. 16; A. serpentina {square with filled S-E corner}, 2n=12, X1 X1 X2 X2 + IVII Meta n. f. 20, {black star}, 2n=11 X1 X2, Yg+IVII Meta n. f.22. It is proposed that this group of species has Robertsonian translocations as a speciation mechanism parting from A. serpentina.


Meza J.S.,Campana Nacional Moscas de la Fruta | Nirmala X.,U.S. Department of Agriculture | Zimowska G.J.,U.S. Department of Agriculture | Zepeda-Cisneros C.S.,Campana Nacional Moscas de la Fruta | Handler A.M.,U.S. Department of Agriculture
Genetica | Year: 2011

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. © 2010 Springer Science+Business Media B.V.


Bosa C.F.,Colegio de Mexico | Cruz-Lopez L.,Colegio de Mexico | Zepeda-Cisneros C.S.,Campana Nacional Moscas de la Fruta | Valle-Mora J.,Colegio de Mexico | And 2 more authors.
Insect Science | Year: 2016

We compared the calling and mating behavior and volatile release of wild males Anastrepha ludens (Loew) with males from 4 mass-reared strains: (i) a standard mass-reared colony (control), (ii) a genetic sexing strain (Tap-7), (iii) a colony started from males selected on their survival and mating competitiveness abilities (selected), and (iv) a hybrid colony started by crossing wild males with control females. Selected and wild males were more competitive, achieving more matings under field cage conditions. Mass-reared strains showed higher percentages of pheromone calling males under field conditions except for Tap-7 males, which showed the highest percentages of pheromone calling males under laboratory cage conditions. For mature males of all strains, field-cage calling behavior increased during the last hour before sunset, with almost a 2 fold increase exhibited by wild males during the last half hour. The highest peak mating activity of the 4 mass-reared strains occurred 30 min earlier than for wild males. By means of solid phase microextraction (SPME) plus gas chromatography-mass spectrometry (GC-MS), the composition of volatiles released by males was analyzed and quantified. Wild males emitted significantly less amounts of (E,E)-α-farnesene but emitted significantly more amounts of (E,E)-suspensolide as they aged than mass-reared males. Within the 4 mass-reared strains, Tap-7 released significantly more amounts of (E,E)-α-farnesene and hybrid more of (E,E)-suspensolide. Differences in chemical composition could be explained by the intrinsic characteristics of the strains and the colony management regimes. Characterization of calling behavior and age changes of volatile composition between wild and mass-reared strains could explain the differences in mating competitiveness and may be useful for optimizing the sterile insect technique in A. ludens. © 2016 Institute of Zoology, Chinese Academy of Sciences.


PubMed | Campana Nacional Moscas de la Fruta
Type: Journal Article | Journal: Genetica | Year: 2011

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 G(0) 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, As2-tub-DsRed.T3], has the DsRed.T3 fluorescent protein reporter gene regulated by the A. suspensa As2-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.


PubMed | Colegio de Mexico and Campana Nacional Moscas de la Fruta
Type: Journal Article | Journal: Insect science | Year: 2016

We compared the calling and mating behavior and volatile release of wild males Anastrepha ludens (Loew) with males from 4 mass-reared strains: (i) a standard mass-reared colony (control), (ii) a genetic sexing strain (Tap-7), (iii) a colony started from males selected on their survival and mating competitiveness abilities (selected), and (iv) a hybrid colony started by crossing wild males with control females. Selected and wild males were more competitive, achieving more matings under field cage conditions. Mass-reared strains showed higher percentages of pheromone calling males under field conditions except for Tap-7 males, which showed the highest percentages of pheromone calling males under laboratory cage conditions. For mature males of all strains, field-cage calling behavior increased during the last hour before sunset, with almost a 2 fold increase exhibited by wild males during the last half hour. The highest peak mating activity of the 4 mass-reared strains occurred 30 min earlier than for wild males. By means of solid phase microextraction (SPME) plus gas chromatography-mass spectrometry (GC-MS), the composition of volatiles released by males was analyzed and quantified. Wild males emitted significantly less amounts of (E,E)--farnesene but emitted significantly more amounts of (E,E)-suspensolide as they aged than mass-reared males. Within the 4 mass-reared strains, Tap-7 released significantly more amounts of (E,E)--farnesene and hybrid more of (E,E)-suspensolide. Differences in chemical composition could be explained by the intrinsic characteristics of the strains and the colony management regimes. Characterization of calling behavior and age changes of volatile composition between wild and mass-reared strains could explain the differences in mating competitiveness and may be useful for optimizing the sterile insect technique in A. ludens.

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