Objective
The main objectives of the project are:
* To develop versatile vectors for transgenesis of Tephritid diptera, based on the transposable elements that have already been used for germ line transformation of other non-Drosophila insects.
* To identify dominant genetic markers appropriate for detection and study of the transgenes.
* To genetically transform at least one fruit fly species of the Bactrocera dorsalis complex with one or more transposon-based vectors and the appropriate markers.
* To assess the genetic stability of the B. dorsalis transgenes under conditions of mass culture in the laboratory.
Expected Outcome
The work carried out in this project should result in the optimization of existing technology and the development of new technologies for germ line transformation of Tephritids of economic importance. It should identify new transposon vectors for stable transgenesis and dominant genetic markers for identification and assessment of the transgenes. The project should lead to the production of several transgenes of B. dorsalis, an agricultural pest of great importance for SE Asia, and the assessment of the stability of these transgenes under mass culture conditions.
The activities envisaged are:
* Assessment of stability of existing fruitfly transgenes. Transgenic Medfly (C. capitata) strains, each carrying a single, independent chromosomal insertion of a Minos transposon marked with the white dominant eye color gene in a white mutant genetic background will be used. Phenotypic reversion or modification will be followed for about twenty generations in population cages.
* Characterization of endogenous transposable elements in two model species, C. capitata and B. tryoni, and in species of the B. dorsalis complex. Using PCR, sequences related to hobo, Minos, and mariner will be identified in the genomes of the three species. PCR fragments will be used as probes to verify their genomic presence and copy numbers. Genomic clones and/or cloned PCR fragments obtained will be sequenced in order to identify full-length and functional elements.
* Identification and characterization of dominant, selectable or visible genetic markers for transformation. The white eye gene (w) that has been used successfully for germ line transformation of the Medfly is potentially useful for other fruit flies, provided that they carry mutations of the w gene. Because a w mutant in B. dorsalis may not be available, the usefulness of other markers, such as alcohol dehydrogenase and GFP, will be investigated in the model species C. capitata.
* Development of a stable, high frequency transformation system for B. dorsalis. Transposable element mobility assays will be used to determine which of the transposons are mobile in B. dorsalis in the presence of the corresponding transposase, but stable in its absence. Transformation will then be attempted using the appropriate transposons with the appropriate genetic markers.
* Assessment of B. dorsalis transgene stability under mass-culture conditions. This will be done by following the stability of inheritance of the genetic marker used for transgenesis.
Fields of science (EuroSciVoc)
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques. See: The European Science Vocabulary.
CORDIS classifies projects with EuroSciVoc, a multilingual taxonomy of fields of science, through a semi-automatic process based on NLP techniques. See: The European Science Vocabulary.
- agricultural sciences agriculture, forestry, and fisheries agriculture horticulture fruit growing
- natural sciences biological sciences genetics mutation
- natural sciences chemical sciences organic chemistry alcohols
- natural sciences biological sciences zoology entomology
- natural sciences biological sciences genetics genomes
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Coordinator
71110 IRAKLION
Greece
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