A Novel Genetic Sexing Strain of Anastrepha ludens for Cost-Effective Sterile Insect Technique Applications: Improved Genetic Stability and Rearing Efficiency.

Anastrepha ludens (Loew) is one of the most destructive insect pests damaging several fruits of economic importance. The sterile insect technique (SIT) is used under an area-wide integrated pest management approach, to suppress these pest populations. Mass rearing facilities were initially established to produce sterile males of bi-sexual strains in support of SIT. The first genetic sexing strain (GSS) for A. ludens, Tapachula-7, based on pupal color dimorphism, was a key development since the release of males-only significantly increases the SIT efficiency. In this study, we document the development of a novel pupal color-based GSS. Twelve radiation-induced translocation lines were assessed as potential GSS in terms of recombination rates and rearing efficiency at a small scale. The best one, GUA10, was cytogenetically characterized: it was shown to carry a single translocation between the Y chromosome and chromosome 2, which is known to carry the black pupae marker. This GSS was further evaluated at medium and large scales regarding its genetic stability, productivity and quality versus Tapachula-7. GUA10 presented better genetic stability, fecundity, fertility, production efficiency, flying ability, and male mating, clear indicators that GUA10 GSS can significantly improve the efficacy and cost-effectiveness of SIT applications against this pest species.

Evaluation of horizontal gene transfer risk between the Mediterranean fruit fly Ceratitis capitata (Tephritidae) and its parasitoid Fopius ceratitivorus (Braconidae).

The transgenic strain of the Mediterranean fruit fly (medfly), Ceratitis capitata (Wied.) VIENNA 8 1260, developed from the classical genetic sexing strain VIENNA 8, has two molecular markers that exhibit red fluorescence in the body and green fluorescence in testicles and sperm. These traits offer a precise tool to discriminate between mass-reared sterile males and wild fertile males, and they could potentially increase the effectiveness of control programs for this pest. To assess the risk of horizontal transfer of the fluorescence transgenes in natural ecosystems, we used the VIENNA 8 1260 strain and the medfly parasitoid Fopius ceratitivorus. The fluorescence signal and the inheritance of the fluorescence gene markers were monitored for over 16 generations (about two years) in both species using fluorescence microscopy and a PCR-based assay. The PCR analysis was performed in four independent laboratories. Both fluorescence microscopy and PCR analysis indicated that no horizontal gene transfer of the DsRed transgene occurred during 16 generations of medfly parasitoid rearing under experimental conditions.

Effect of Irradiation Doses on Sterility and Biological Security in a Genetically Modified Strain of the Mediterranean Fruit Fly (Diptera: Tephritidae).

The genetically modified strain of the Mediterranean fruit fly, Ceratitis capitata (Wiedemann) VIENNA 8 1260, was developed from the genetic sexing strain VIENNA 8. It has two molecular markers that exhibit red fluorescence in the body and green fluorescence in testis and sperm. These traits offer a precise tool to discriminate between mass-reared and wild males, increasing the effectiveness of sterile insect technique. The reproductive performance of the VIENNA 8 1260 and VIENNA 8 D53- (without the D53 inversion introduced to prevent recombination) was compared at different irradiation doses. The general effect of irradiation on VIENNA 8 1260 followed the same patterns documented in previous publications for VIENNA 8 D53-. Irradiation doses of 80 Gray or greater reduced fertility and induced high levels of sterility in wild females. Fecundity reduction was higher in VIENNA 8 1260 than in VIENNA 8 D53- females. Vertical transmission of the fluorescence gene was confirmed up to the F4 generation. Substerilization in the VIENNA 8 1260 could jeopardize the usefulness of the transgenic strain due to the possible vertical transfer of the fluorescence transgene from the sterile males to the wild flies. A biologically safe higher irradiation dose could result in reduced competitiveness of the VIENNA 8 1260 strain. Mating and remating experiments suggest that Mediterranean fruit fly females exhibit a relative precedence in the use of the sperm: though both sperms are mixed, sperm from the remating is spent first. Results suggest a lower fitness of VIENNA 8 1260 sperm, when compared with sperm from a nonfluorescent bisexual strain, which is consistent with the lower reproductive performance documented for the VIENNA 8 1260 strain.

Performance of a Genetically Modified Strain of the Mediterranean Fruit Fly (Diptera: Tephritidae) for Area-Wide Integrated Pest Management With the Sterile Insect Technique.

The genetically modified strain of Ceratitis capitata (Wiedemann) VIENNA 8 1260 has two morphological markers that exhibit fluorescence in body and sperm. To assess the feasibility of its use in area-wide integrated pest management (AW-IPM) programs using the sterile insect technique, its rearing performance and quality control profile under small, medium, and large scales was evaluated, as well as in field cages. The VIENNA 8 1260 strain had a lower yield than the control strains, VIENNA 8 with D53 inversion (VIENNA 8) and without D53 inversion (VIENNA 8 D53-). At mass-rearing scale, yield gradually increased in three generations without reaching the control strain values. The VIENNA 8 1260 strain was stable in the genetic sexing mechanism (>99.9%) and expression of fluorescence (100%). In field cages, the VIENNA 8 1260 males reduced the mating potential of wild males in the same magnitude as the VIENNA 8, when evaluated in independent cage tests. However, the relative sterility index and the strain male relative performance index of VIENNA 8 1260 males were significantly lower than those of the VIENNA 8. There were no significant differences in longevity of these strains. The potential application of the VIENNA 8 1260 in AW-IPM programs is further discussed.