Genetic Sexing Strains of Mediterranean Fruit Fly (Diptera: Tephritidae): Quality in Mass-Reared Temperature-Sensitive Lethal Strains Treated at High Temperatures

Abstract Improvements in the mass rearing of Diachasmimorpha longicaudata (Ashmead) on larvae of the Vienna-8 temperature-sensitive lethal genetic sexing strain of Ceratitis capitata (Wiedemann) (Diptera: Tephritidae) (= GSS Vienna-8) at the San Juan biofactory, Argentina, are currently under way. Lowering cost production is a key factor regarding parasitoid rearing. Thus, the variation in mass-reared parasitoid encapsulation levels and the incidence of superparasitism were determined; also, the gamma radiation dose-effect relation on host larvae and the influence of Mediterranean fruit fly strain were considered. Naked Mediterranean fruit fly larvae of both GSS Vienna-8 and a wild bisexual strain (= WBS) aged 6-d-old were irradiated at 0, 20, 40, 60, 80, 100, and 120 Gy, and exposed to parasitoid females. Melanization level was tested for encapsulated parasitoid larval first-instars (= L1). Non-irradiated and irradiated WBS larvae at 20–40 Gy displayed a significantly higher incidence of encapsulation when compared with GSS Vienna-8 larvae. The low melanized level in encapsulated parasitoid L1 was the most common melanization process at 72 h puparium dissection. A high melanized level was only found in non-irradiated WBS larvae. Irradiated GSS Vienna-8 larvae can neutralize the host immunological reactions over irradiated WBS larvae much more quickly. Superparasitism intensity in both Mediterranean fruit fly strains was not affected by radiation doses. High levels of superparasitism seemingly helped to overcome the host’s immune reaction by the surviving parasitoid larva. Parasitoid emergence increased from 60 Gy onwards in both Mediterranean fruit fly strains. Radiation in GSS Vienna-8 larvae may favor host’s antagonistic reactions decrease in relation with D. longicaudata development.

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Lessons from Drosophila: Engineering Genetic Sexing Strains with Temperature-Sensitive Lethality for Sterile Insect Technique Applications

Insects ◽

10.3390/insects12030243 ◽

2021 ◽

Vol 12 (3) ◽

pp. 243

Author(s):

Thu N. M. Nguyen ◽

Amanda Choo ◽

Simon W. Baxter

Keyword(s):

Rna Polymerase Ii ◽

Sterile Insect Technique ◽

Genetic Research ◽

Point Mutations ◽

Random Mutagenesis ◽

Fruit Fly ◽

Temperature Sensitive ◽

Agricultural Pests ◽

Genetic Sexing ◽

Genetic Sexing Strains

A major obstacle of sterile insect technique (SIT) programs is the availability of robust sex-separation systems for conditional removal of females. Sterilized male-only releases improve SIT efficiency and cost-effectiveness for agricultural pests, whereas it is critical to remove female disease-vector pests prior to release as they maintain the capacity to transmit disease. Some of the most successful Genetic Sexing Strains (GSS) reared and released for SIT control were developed for Mediterranean fruit fly (Medfly), Ceratitis capitata, and carry a temperature sensitive lethal (tsl) mutation that eliminates female but not male embryos when heat treated. The Medfly tsl mutation was generated by random mutagenesis and the genetic mechanism causing this valuable heat sensitive phenotype remains unknown. Conditional temperature sensitive lethal mutations have also been developed using random mutagenesis in the insect model, Drosophila melanogaster, and were used for some of the founding genetic research published in the fields of neuro- and developmental biology. Here we review mutations in select D. melanogaster genes shibire, Notch, RNA polymerase II 215kDa, pale, transformer-2, Dsor1 and CK2α that cause temperature sensitive phenotypes. Precise introduction of orthologous point mutations in pest insect species with CRISPR/Cas9 genome editing technology holds potential to establish GSSs with embryonic lethality to improve and advance SIT pest control.

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Improved stability of genetic sex-separation strains for the Mediterranean fruit fly, Ceratitis capitata

Genome ◽

10.1139/g94-009 ◽

1994 ◽

Vol 37 (1) ◽

pp. 72-82 ◽

Cited By ~ 58

Author(s):

G. Franz ◽

E. Gencheva ◽

Ph. Kerremans

Keyword(s):

Ceratitis Capitata ◽

Recombination Frequency ◽

Fruit Fly ◽

Mass Rearing ◽

Temperature Sensitive ◽

Genetic Sexing ◽

Male Recombination ◽

Comparable Level ◽

Improved Stability ◽

Genetic Sexing Strains

In the existing genetic sexing strains for the medfly, Ceratitis capitata, male recombination leads to breakdown of the sexing mechanism under mass rearing conditions. The rate of breakdown depends on the recombination frequency and on the fitness of the recombinants. We have tested two different sexing genes, white pupa and a temperature sensitive lethal, in combination with the translocation T(Y;5)30C. Both sexing strains broke down, although at very different rates. In the case of the white pupa strain, 3.5% recombinants were observed after rearing the strain for 15 generations. The second strain, utilizing white pupa and the temperature sensitive lethal as selectable markers, already reached a comparable level after six generations and was broken down completely in the ninth generation. In these strains the frequency of recombination is high because the breakpoint of T(Y;5)30C and the sexing gene(s) are far apart. To remedy the situation, we have isolated four new translocations with breakpoints located closer to the sexing genes. Mass rearing was simulated for several generations with strains based on these translocations and no breakdown was observed under the conditions used.Key words: medfly, sterile insect technique, genetic sexing, recombination.

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