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MOLECULAR BIOLOGY and GENOMICS

 

         Selections
1. Brief Description 6. Thesis
2. Research Interests 7. Photos
3. Personnel 8. Useful links
4. Funded Research Projects 9. Contact
5. Publications    

1. Brief Description:

The revolution in Molecular Biology that took place during the last quarter of the 20th century led to large scale technologies and whole genome analyses, i.e., to the era of Genomics. These approaches are applied in the Laboratory of Molecular Biology and Genomics in our attempt to address crucial questions that regard the biology, ecology and sex differentiation of the major olive pest, the olive fly, in an effort to develop novel, environmentally friendly methods to control this insect.

2. Research interests:

1. Molecular and Biochemical studies of insecticide resistance mechanisms 
The extensive and non-prudent use of chemical insecticides for insect control has lead to the development of resistance. Our aim is to understand the mechanisms of insecticide resistance through the isolation and study of resistance-associated mutations. We have recently discovered a new mutation in the carboxyl-terminal of the acetylcholinesterase gene that indicates a completely novel mechanism of resistance to organophosphate insecticides. We also study resistance development to spinosad, a new insecticide of the naturalyte class. This insecticide is very toxic towards its insect targets, whereas it is relatively innocuous towards beneficial insects, man and the environment. Its molecular target is the nicotinic acetylcholine receptor (nAChR), however the exact resistance mechanism remains unknown. In our effort to discover spinosad resistance associated mutations we have isolated several nAChR subunits. Mutations in these subunits may indicate the points where spinosad binds to the nAChR and shed light to the molecular mechanism of spinosad resistance. 

2. Genomics, transcriptomics and proteomics analysis of the olive fly 
Despite the great economic impact that the olive fly has on all olive producing Mediterranean countries, relatively little is known about its genetics and molecular biology. The development of genetic and molecular tools is important not only for the basic research of the insect, but also for the optimization of biological control methods, such as the sterile insect technique (SIT). Towards this direction, we construct cytogenetic maps marked with molecular markers (microsatellites, ESTs etc). Such maps predate any serious analysis of genome organization, gene characterization, genetic analysis of monogenic and quantitative trait loci, as well as efforts of genomics analysis. We also study transposable elements and other repeats in order to understand chromosome organization, with emphasis on the Y chromosome. Lastly, we develop a basic transcriptomics (Serial Analysis of Gene Expression) and Proteomics approach in order to isolate genes that are involved either in sex-separation pathways or resistance to insecticides. 

3. Invasion and adaptation of olive fly populations
Olive production has a century-old history in California. However, the olive fly was first detected in the Los Angeles area in 1998 and within five years it was reported from all olive cultivation areas of the state. Analysis of several molecular markers (microsatellite, mtDNA, rRNA and Y-specific markers) of California field-collected flies pointed to the eastern part of the Mediterranean as the putative source of the observed invasion. We also study the possibility of multiple olive fly invasions in the state of California, as well as the question of adaptation of natural populations in a new environment. This last question is also addressed in laboratory conditions: what are the changes of microsatellite allele frequencies during colonization in the laboratory? What does fixation or disappearance of certain alleles tell us about the neutrality of the molecular marker used or of its neighbouring sequences?

 

3. Personnel:

Principal Investigator
Kostas Mathiopoulos

Members

KonstantinaTsoumani, Post Doctoral Research Associate, University Fellow as teaching Lecturer
Nikos Zygouridis, PhD Candidate
EfthimiaSagri,PhD Candidate
Maria-EleniGregoriou, PhD Candidate
PanagiotaKoskinioti, PhD Candidate
Belavillas-TrovasAlexandros, Undergraduate Student


4. Funded Research Projects:

  • 2006-2008: Bilateral Greece-Cyprus R&T Collaborative Programmes of the General Secretariat of Research and Technology of the Ministry of Development, Greece, on "Assessment of resistance of natural olive fly populations to the naturalyte insecticide spinosad as well as investigation of the underlying molecular mechanism of such resistance".
  • 2005-2007: Research Potential Support Programme of the General Secretariat of Research and Technology of the Ministry of Development, Greece, on "Assessment of insecticide resistance in four insects of great economic importance (olive fly, green peach aphid, medfly and apple maggot fly), investigation of underlying resistance mechanisms and study of their bio-ecology.
  • 2005-2006: University Research Teams Support Programme "Pythagoras ΙΙ" of the Greek Ministry of National Education and Religious Affairs on: «Molecular analysis of olive fly chromosomes, with emphasis on the Y chromosome, as well as use of molecular markers for the assessment of the fly's invasion in California».
  • 2004-2006: Bilateral Greece-Cyprus R&T Collaborative Programmes of the General Secretariat of Research and Technology of the Ministry of Development, Greece, on "Assessment of organophosphate and pyrethroid resistance of the olive fly".
  • 2009 - today   Coordinated Research Project of International Atomic Energy Agency entitled: "Assessment of Genetic Diversity of the Olive Fly Israeli SIT Laboratory Strain".
  • 2010-2012: Specialty Crops Block Grant Program from California Department of Food and Agriculture on: «Spinosad Resistance in California Olive Fruit Fly (Bactrocera oleae) Populations».
  • 2010-2012: "Targeted Research Actions" Programmes of the University of Thessaly Research Committee, entitled: "HPV incidence in the upper respiratory system of the healthy human population of Thessaly".
  • 2010-2013: Research Potential Support Programme Heraclitus II of the Greek Ministry of Education on: "Transcriptomics and proteomics analysis of the most important olive pest, the olive fly Bactrocera oleae, focusing on sex-separation and insecticide resistance systems".
  • 2013-2014: "Sexing strains based on embryonic lethality for SIT in olive fly", financed by the Programme for the promotion of the exchange and scientific cooperation between Greece and Germany (IKYDA 2013).
  • 2012-2015: "Novel approaches to Olive fly control: focus on olfactory and reproductive systems", implemented under the "ARISTEIA" Action of the "OPERATIONAL PROGRAMME EDUCATION AND LIFELONG LEARNING" and co-funded by the European Social Fund (ESF) and National Resources.
  • 2012-2014: Interdisciplinary and Interuniversity Research Support Program of the Greek Ministry of Education "Thales" entitled: "Symbiotic bacteria and 'omics' technologies towards the development of novel and environment-friendly control methods on insect pests: the case of the Mediterranean fruit fly". Participation as Principal Investigator of Collaborating Team.



5. Publications:

  1. Stratikopoulos EE, Augustinos AA, Gariou-Papalexiou A, Zacharopoulou A and Mathiopoulos KD (2002). Identification and partial characterization of a new Ceratitis capitata specific 44-bp centromeric repeat. Chromosome Res 9: 287-295. http://www.ncbi.nlm.nih.gov/pubmed/12199142
  2. Augustinos AA, Stratikopoulos EE, Zacharopoulou A and Mathiopoulos KD (2002). Polymorphic microsatellite markers in the olive fly, Bactrocera oleae. Mol Ecol Notes 2: 278-280. http://onlinelibrary.wiley.com/doi/10.1046/j.1471-8286.2002.00222.x/abstract
  3. Augustinos AA, Mamuris Z, Stratikopoulos EE, D’Amelio S, Zacharopoulou A and Mathiopoulos KD (2005). Microsatellite analysis of olive fly populations in the Mediterranean indicates a westward expansion of the species. Genetica 125: 231-241. http://www.ncbi.nlm.nih.gov/pubmed/16247695
  4. Skouras JP, Margaritopoulos JT, Seraphides NA, Ioannides IM, Kakani EG, Mathiopoulos KD and Tsitsipis JA (2007). Organophosphate resistance in olive fly, Bactrocera oleae, populations in Greece and Cyprus. Pest Manag Sci 63: 42-48. http://www.ncbi.nlm.nih.gov/pubmed/17103369
  5. Stratikopoulos EE, Augustinos AA, Petalas YG, Vrahatis MN, Mintzas A, Mathiopoulos KD and Zacharopoulou A (2008). An integrated genetic and cytogenetic map for the Mediterranean fruit fly, Ceratitis capitata, based on microsatellite and morphological markers. Genetica 133: 147-157. http://www.ncbi.nlm.nih.gov/pubmed/17786564
  6. Kakani EG, Ioannides IM, Margaritopoulos JT, Seraphides NA, Skouras PJ, Tsitsipis JA, Mathiopoulos KD (2008). A small deletion in the olive fly acetylcholinesterase gene associated with high levels of organophosphate resistance. Insect Biochem Mol Biol 38: 781-787. http://www.ncbi.nlm.nih.gov/pubmed/18625401
  7. Zygouridis NE, Augustinos AA, Zalom FG and Mathiopoulos KD (2009). Analysis of Olive Fly Invasion in California Based on Microsatellite Markers. Heredity 102: 402-412. http://www.ncbi.nlm.nih.gov/pubmed/19107137
  8. Kakani EG and Mathiopoulos KD (2008). Organophosphate resistance-related mutations in the acetylcholinesterase gene of Tephritidae. J Appl Entomol 132: 762-771. http://onlinelibrary.wiley.com/doi/10.1111/j.1439-0418.2008.01373.x/abstract
  9. Augustinos AA, Stratikopoulos EE, Drosopoulou E, Kakani EG, Mavragani-Tsipidou P, Zacharopoulou A, Mathiopoulos KD (2008). Isolation and characterization of microsatellite markers from the olive fly, Bactrocera oleae, and their cross-species amplification in the Tephritidae family. BMC Genomics 9(1): 618. http://www.ncbi.nlm.nih.gov/pubmed/19099577
  10. Liakopoulos A, Neocleous C, Klapsa D, Kanellopoulou M, Spiliopoulou I, Mathiopoulos KD, Papafrangas E and Petinaki E. A T2504A mutation in the 23S rRNA gene responsible for high-level resistance to linezolid of Staphylococcus epidermidis. J Antimicrob Chemother. 2009 May 09. http://www.ncbi.nlm.nih.gov/pubmed/19429927
  11. Stratikopoulos EE, Augustinos AA, Pavlopoulos I, Economou K, Mintzas A, Mathiopoulos KD and Zacharopoulou A (2009). Isolation and characterization of microsatellite markers from the Mediterranean fruit fly, Ceratitis capitata: cross-species amplification in other Tephritidae species reveals a varying degree of transferability. Mol Genet Genomics 282(3): 283-306. http://www.ncbi.nlm.nih.gov/pubmed/19544072
  12. Papagiannoulis A, Mathiopoulos KD, Mossialos D (2009). Molecular detection of the entomopathogenic bacterium Pseudomonas entomophila using PCR. Lett Appl Microbiol 50(3): 241-245. http://www.ncbi.nlm.nih.gov/pubmed/20025650
  13. Kakani EG, Zigouridis NE, Tsoumani K, Seraphides N, Zalom FG and Mathiopoulos KD (2010). Spinosad resistance development in wild olive fruit fly Bactrocera oleae (Diptera: Tephritidae) populations in California. Pest Manag Sci 66(4):447-453. http://www.ncbi.nlm.nih.gov/pubmed/20146256
  14. Tsoumani KT, Augustinos AA, Kakani EG, Drosopoulou E, Mavragani-Tsipidou P and Mathiopoulos KD (2011) Isolation, annotation and applications of expressed sequence tags from the olive fly, Bactrocera oleae. Mol Genet Genomics 285: 33-45. http://www.ncbi.nlm.nih.gov/pubmed/20978910
  15. Kakani EG, Bon S, Massoulié J and Mathiopoulos KD (2011) Altered GPI modification of insect AChE improves tolerance to organophosphate insecticides. Insect Biochem Mol Biol 41: 150-158 http://www.ncbi.nlm.nih.gov/pubmed/21112395
  16. Vontas J, Hernández-Crespo P, Margaritopoulos JT, Ortego F, Feng H-T, Mathiopoulos KD, Hsu J-H (2011) Insecticide resistance in Tephritid flies. PesticBiochem Physiol100: 199-205 http://www.sciencedirect.com/science/article/pii/S004835751100068X
  17. Tsoumani KT and Mathiopoulos KD (2011) Genome size estimation with quantitative real-time PCR in two Tephritidae species: Ceratitis capitata and Bactrocera oleae. J Appl Entomol 136: 626-631 http://onlinelibrary.wiley.com/doi/10.1111/j.1439-0418.2011.01684.x/abstract
  18. Kakani EG, Trakala M, Drosopoulou E, Mavragani-Tsipidou P and Mathiopoulos KD (2012) Genomic structure, organization and localization of the acetylcholinesterase locus of the olive fruit fly, Bactroceraoleae. Bull Entomol Res 12: 1-12 http://journals.cambridge.org/action/displayAbstract?fromPage=online&aid=8812699
  19. Zygouridis NE, Argov Y, Nemny-Lavy EE, Augustinos AA, Nestel D and Mathiopoulos KD (2013) Genetic changes during laboratory domestication of an olive fly SIT strain. J Appl Entomol (In press) http://onlinelibrary.wiley.com/doi/10.1111/jen.12042/abstract;jsessionid=1B3CFE349045718796472A556F197C0B.f03t01
  20. Kakani EG, Sagri E, Omirou M, Ioannides IM and Mathiopoulos KD (2013) Detection and geographical distribution of the organophosphate resistance-associated Δ3Q ace mutation in the olive fly, Bactrocera oleae (Rossi). Pest Manag Sci. (In press) http://www.ncbi.nlm.nih.gov/pubmed/23908134
  21. Tsoumani KT, Drosopoulou E, Mavragani-Tsipidou P and Mathiopoulos KD (2013). Molecular characterization and chromosomal distribution of a species-specific centromeric repetitive DNA sequence from the olive fruit fly, Bactrocera oleae (Rossi). PLoS One (In press). http://www.ncbi.nlm.nih.gov/pubmed/24244494

 

6. Thesis:

Undergraduate
  1. Evdoxia Kakani (2000) EST marker analysis in the olive fly, Bactrocera oleae.
  2. Panagiotis Giannios (2000) Molecular identification of Anisakis species in marine mammals in the Gulf of Mexico.
  3. Agoritsa Kaliora (2000) Molecular identification of Echinococcus granulosus strains in infected samples from Italy, Slovakia, Argentina and Portugal.
  4. Konstantina Tsoumani (2001) Physical map construction in the olive fly, Bactrocera oleae.
  5. Sophia Maiou (2001) Analysis of natural populations of the olive fly.
  6. Efstathia Kosmidou (2001) Cloning of the GST genetic locus, involved in insecticide resistance of the olive fly, Bactocerae οleae.
  7. Marianna Trakala (2001) The protective action of melatonin in mitochondria during sepsis.
  8. Giorgos Efstathiou (2002) Cloning of the α-esterase genetic locus in the Medfly, Ceratitis Capitata (Wiedemann).
  9. Barbara Podia (2002) Detection of insecticide resistance associated mutations in the acetylcholinesterase locus of the olive fly.
  10. Irene Papadaki (2002) Expression of the extracellular fragment of the nicotinic acetylcholine receptor A1 subunit in E. coli strains.
  11. Christina Massia (2002) Polymorphism analysis of the acetylcholinesterase locus in natural olive fly populations in Cyprus.
  12. Pavlos Panagi (2002) Analysis of organophosphate resistance mutations in natural olive fly populations from Cyprus.
  13. Ourania Trochatou (2003) Contribution of retrotransposons and repetitive elements in the molecular organization of the olive fly chromosomes.
  14. Katerina Katechou (2004) (in progress) Isolation and analysis of Achilles-containing phage clones, an olive fly retrotransposable element.
  15. Katerina Mitsiou (2004) Study of the possibility of multiple olive fly invasions in California with the use of microsatellite markers.
  16. Elli Ksiromeriti (2004) Insertion polymorphisms of Achilles, an olive fly retrotransposon.
  17. Irene Chainoglou (2004) Cloning of nicotinic acetylcholine receptor subunits of the olive fly, Bactrocera oleae, involved in the resistance against the naturalyte insecticide spinosad.
  18. Antonia Chroni (2004) Sex detection in the olive fly, Bactrocera oleae, with the use of molecular markers.
  19. Panagiota Koskinioti (2005) Molecular genotyping of Leishmania parasites in the canine population of Thessaly.
  20. Eleni Seraphimidou-Pouliou (2005) Study of the adaptation of olive fly populations in laboratory conditions.
  21. Anastasia Asimakopoulou (2005) Development and partial analysis of microsatellite markers in the cherry fly, Rhagoletis cerasi.
  22. Charalabos Kontopoulos (2005) Population analysis of olive fly natural populations by SSCP analysis.
  23. Aggeliki Kiriazou (2005) Molecular genotyping of Leishmania parasites in the human population of Thessaly.
  24. Panagiota Kimonos (2005) Molecular analysis of a family with autosomal Hereditary Inclusion Body Myopathy.
  25. Maria-Eleni Gregoriou (2005) Study of the molecular mechanism of resistance against the naturalyte insecticide spinosad in the olive fly, Bactrocera oleae.
  26. Kristallo Christou (2006) Targeted silencing of Tephritidae genes, using the RNAi technology in the fly food.
  27. Nikos Manos (2006) Targeted silencing of Medfly genes with the use of RNAi technology in the larval food.
Graduate
  1. Efthimia Sagri (2007) Spread of organophosphate resistance-associated mutations in olive fly populations
  2. Eleni Gema (2007) Development of diagnostic molecular markers in natural populations of the olive fly
  3. Athanasios Tsagkadopoulos (2007) Diachronic population dynamics of the olive fly invasion in California
  4. Niki Lourou (2008) Study of the spread of insecticide resistance alleles in olive fly populations
  5. Politimi Kitsidi (2008) Sexual interaction of zygomycete Phycomyces blakesleeanus. Isolation, structural characterization and biosynthesis of trisporoids and derivatives
  6. Titika Kontaksi (2008) Sexual interaction of zygomycete Blakeslea trispora. Isolation, structural characterization and biosynthesis of trisporoids and derivatives
  7. Paschalis Chlopsides (2009) Molecular analysis of Leishmania haplotypes in the canine population of Thessaly
PhD
  1. Evdoxia Kakani (2005) Molecular analysis of insecticide resistance loci in the olive fly, Bactrocera oleae
  2. Konstantina Tsoumani (2006) Analysis of the molecular organization of the sex determining chromosome Y of the olive fly, Bactrocera oleae
  3. Nikos Zygouridis (2007) Population analysis of the olive fly, Bactrocera oleae, with the use of molecular markers 
  4. Efthimia Sagri (2009) Transcriptomics and proteomics analysis of the olive fly, Bactrocera oleae, with emphasis on sex separation systems and insecticide resistance
 

 

7. Photos:

 

 

8. Useful Links:

 

9. Contact:

Kostas Mathiopoulos: This e-mail address is being protected from spambots. You need JavaScript enabled to view it

University of Thessaly
Department of Biochemistry & Biotechnology
Viopolis
Larissa – 41500
Greece

Αntonios Giakountis:  This e-mail address is being protected from spambots. You need JavaScript enabled to view it

University of Thessaly
Department of Biochemistry & Biotechnology
Viopolis
Larissa – 41500
Greece

Konstantina Tsoumanii: This e-mail address is being protected from spambots. You need JavaScript enabled to view it

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