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Identifier 000414790
Title Immunolocalizationstudy of detoxification enzymes (Cytochrome P450, Glutathione S-Transferase, Intradiol Ring- CleavageDioxygenase) in themajor agricultural pest Tetranychus urticae
Alternative Title Μελέτη ανοσοεντοπισμού ενζύμων αποτοξικοποίησης (Κυτοχρωμικές P450, Μεταφοράσες της γλουταθειόνης, Διοξυγενάσες) στο άκαρι Tetranychus urticae
Author Παπαδάκη, Σταυρινή
Thesis advisor Βόντας, Ιωάννης
Τζαμαρίας, Δημήτριος
Abstract Tetranychus urticae Koch, 1836 is a major agricultural pest that belongs to the Subclass of Acari. It is known for its polyphagous behavior, as it can infest more than 1.100 different plant species, as well as its ability to develop resistance to insecticides/acaricides in very short periods after their first application. Some of its biological characteristics contribute to resistance development such as its high fecundity, haplo-diploid sex determination and short life cycle. Additional characteristics such as its polyphagous behavior have been correlated with the ability of T. urticae to counteract the effect of insecticides based on the pre-adaptation theory. The sequencing of T.urticae genome further supports this hypothesis since it revealed an extraordinary arsenal of detoxification enzymes, many of which are T. urticae specific. Previous studies have shown the over-expression of members of CYP392 family (CYP392A11, CYP392A12, CYP392A16, CYP392D2, CYP392D8, CYP392D10) in resistant strains with CYP392A11 metabolizing the METI insecticides fenpyroximate and cyenopyrafen and CYP392A16 metabolizing the avermectin insecticide abamectin. Additionally, members of GST family (TuGSTd05, TuGSTd10, TuGSTd14) have been found to be over-expressed in resistant strains with TuGSTd05 possibly metabolizing the METI insecticide cyflumetofen. In this study we aimed in localizing three major enzyme families (CYP392A, GSTd, ID-RCD). The first two are of known function and it has been confirmed that their increased presence confers resistance. The third is a newly discovered family in arthropods and constitutes a compelling case of horizontal gene transfer, with its function being unknown. Additionally, there are several documented cases where the function of ID-RCDs has been correlated with digestion or detoxification. We raised 4 different antibodies (anti-CYP392A, anti-CYP392D, anti-GSTd and anti- Dioxygenase) each of which was first tested on Western blot analyses to see their specificity. Three out of four were not specific for the selected subfamily/family (anti-CYP392D, anti-GSTd and anti- Dioxygenase) so we continued the immunolocalization studies with anti-CYP392A and anti-CYP392A16, which were used in cryo-sections and whole mount specimens of T. urticae adult females. A part of the results using anti-CYP392A16 indicates the possible presence of A16 in nerve cells proximal to the central nervous mass of T. urticae. This finding is in accordance with previously conducted experiments on different organisms such as the mosquito Aedes albopictus (Grigoraki et al. 2016), the silkworm Bombyx mori (Xuan et al. 2014) and the red floor beetle Tribolium castaneum (Zhu et al. 2010). Our efforts should be focused in finalizing the immunolocalization protocol so that our data are consistent, while the co-localization of CPR, the redox partner of CYP392A16, would further support the data.
Language English
Subject CYP392
Issue date 2018-03-23
Collection   Faculty/Department--Faculty of Sciences and Engineering--Department of Biology--Post-graduate theses
  Type of Work--Post-graduate theses
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