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Identifier 000397198
Title Functional expression and characterization of detoxification enzymes in insects and mites
Alternative Title Λειτουργική έκφραση και χαρακτηρισμός ενζύμων αποτοξικοποίησης σε έντομα και ακάρεα
Author Τσέλιου, Βασίλης
Thesis advisor Βόντας, Ιωάννης
Μπουριώτης, Βασίλης
Abstract Studying the enzymes by which agricultural pests develop resistance to insecticides, aiming to develop new means of managing and overcoming this resistance, a crucial task for crop protection. I studied detoxification genes and enzymes, which are associated within the resistance phenotype in two major pests: The two spotted spider mite Tetranychus urticae is one of the most damaging pests in agriculture. T. urticae is extremely polyphagous; it can feed on hundreds of plants including important agricultural crops. Genome wide gene expression analysis (micaroarray) of a multiresistant strain of T.urticae (Marathonas) showed, among others, the association of two Mu class Glutathione S tranferases (GSTs), TuGSTm07 and TuGSTm09, with the resistant phenotype. In this study, both were functionally expressed and kinetically characterized and their potential to interact with insecticides/acaricides were examined. TuGSTm07 found to interact with bifenthrin causing 70% inhibition of its CDNB coguncating activity. The IC50 value was determined at 7,7103 ± 0,956 μM, showing strong interaction. The three dimensional structure of TuGSTm07 was predicted based on X-ray of other Mammals’ Mu GST. Docking with bifenthrin was performed and revealed that bifenthrin most likely binds to the active site of the enzyme. Bactocera oleae (Gmelin) (Diptera: Tephritidae) is the most important insect pest of the olive tree, causing 30 % loss of the olive crop in Mediterranean countries including Greece. The pyrethroid α-cypermethrin is currently used against B. oleae, however high levels of resistance have been recently observed. Recently, after examination of the expression changes in resistant populations compared to susceptible, fourteen genes were found to be commonly up-regulated. Among them, contig00436, putative member of the CYP6 cytochrome P450 family, was identified as the most striking hit. In this study, the overexpression of contig00436 was validated in resistant populations by real time quantitative PCR, cloned into pcW-ompA expression vector and functionally expressed, aiming to investigate if it is capable of metabolizing α-cypermethrin in vitro.
Language English
Issue date 2015-11-20
Collection   Faculty/Department--Faculty of Sciences and Engineering--Department of Biology--Post-graduate theses
  Type of Work--Post-graduate theses
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