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INCO project Artemia biodiversity Beijing Workshop, September 23-26, 2002

INCO project Artemia biodiversity Beijing Workshop, September 23-26, 2002. Artemia characterisation based mainly on mtDNA RFLP analyses and progress in Mediterranean Basin Regional Action A. Triantafyllidis, A.D. Baxevanis, A. Tzika, E. Markatzinou & T.J. Abatzopoulos

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INCO project Artemia biodiversity Beijing Workshop, September 23-26, 2002

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  1. INCO project Artemia biodiversity Beijing Workshop, September 23-26, 2002 Artemia characterisation based mainly on mtDNA RFLP analyses and progress in Mediterranean Basin Regional Action A. Triantafyllidis, A.D. Baxevanis, A. Tzika, E. Markatzinou & T.J. Abatzopoulos ARISTOTLE UNIVERSITY OF THESSALONIKI (AUTH) INCO Partner: 3

  2. Current research in AUTH – Objectives of INCO project • Description of existing gene pools focusing on Artemia populations from: • North and South America • South Africa • Mediterranean Basin • Evolutionary history of parthenogenetic Artemia Molecular markers (identifying clones) Life-history traits • Morphometry Differentiation of populations – Database • Visiting scientists INCO meeting, 23-26 September, Beijing, China Aristotle University of Thessaloniki (AUTH)

  3. Visiting Scientists • University de Los Lagos, Chile (INCO partner 14) • Mr. Julio Crespo • 2,5 months • Rhodes University, South Africa (INCO partner 9) • Dr. Horst Kaiser • 1,5 months INCO meeting, 23-26 September, Beijing, China Aristotle University of Thessaloniki (AUTH)

  4. Molecular techniques used at AUTH • DNA extraction: Phenol extraction, Chelex • All analyses were done at INDIVIDUAL level • PCR amplification of a mitochondrial 16S rRNA region • Restriction Fragment Length Polymorphism (RFLP) analysis with an initial screening of 16restriction enzymes • Analysis of data: REAP, PHYLIP, ARLEQUIN INCO meeting, 23-26 September, Beijing, China Aristotle University of Thessaloniki (AUTH)

  5. Chilean populations analysed in AUTH INCO meeting, 23-26 September, Beijing, China Aristotle University of Thessaloniki (AUTH)

  6. EL CONVENTO LLAMARA CHAXAS CEJAS A. franciscana LOS VILOS A. franciscanaULA A. franciscanaAUTH IQUIQUE A. persimilisAUTH A. persimilisULA A. persimilis TORRES DEL PAINE PICHILEMU 0.01 UPGMA Dendrogram of Chilean Artemia populations INCO meeting, 23-26 September, Beijing, China Aristotle University of Thessaloniki (AUTH)

  7. Species/population specific markers (restriction enzymes) were identified in order to distinguish: 1. A. fransciscana from A. persimilis 2. Within A. fransciscana Northern from southern populations 3. Within A. persimilis Pichilemu from Torres del Paine population INCO meeting, 23-26 September, Beijing, China Aristotle University of Thessaloniki (AUTH)

  8. Populations scored by Dr. H. Kaiser Analysis of parthenogenetic and bisexual populations INCO meeting, 23-26 September, Beijing, China Aristotle University of Thessaloniki (AUTH)

  9. Additional populations characterized at AUTH INCO meeting, 23-26 September, Beijing, China Aristotle University of Thessaloniki (AUTH)

  10. salina kesani mesi urmia kal mad kesani mesi urmia polich salina Results of restriction analyses BfaI AluI INCO meeting, 23-26 September, Beijing, China Aristotle University of Thessaloniki (AUTH)

  11. FRANC1 FRANC2 75 FRANC4 FRANC3 SALINA 70 COEGA1 PARTH1 95 URMIA PARTH4 PARTH2 COEGA2 PARTH3 PERS2 98 PERS3 PERS1 0.01 UPGMA Phylogenetic tree of Artemia haplotypes INCO meeting, 23-26 September, Beijing, China Aristotle University of Thessaloniki (AUTH)

  12. El Convento Llamara Chaxas Cejas Los Vilos A. franciscana ULA A. franciscana AUTH Iquique A. salina Coega1 M. Embolon Madagascar Namibia A. urmiana Kalloni Polychnitos Kessani Mesi Coega2 Citros A. persimilis ULA A. persimilis AUTH Torres del Paine Pichilemu UPGMA Phylogenetic tree of Artemia populations franciscana salina clade A parthen. clade B persimilis 0.01 INCO meeting, 23-26 September, Beijing, China Aristotle University of Thessaloniki (AUTH)

  13. Two major clades were identified within parthenogetic populations • Genetic distance among these clades is similar to the • genetic distance among A. franciscana and A. salina • No evident geographic structure found. • Both clades coexist in nearby or even same site • e.g. Polychnitos • A. urmiana genetic fingerprint identical to one of the two clades INCO meeting, 23-26 September, Beijing, China Aristotle University of Thessaloniki (AUTH)

  14. The Coega population comprises of two different strains: a bisexual and an asexual one. • The bisexual strain is grouped with A. salina • The parthenogenetic strain clusters with one of the two • parthenogenetic clades INCO meeting, 23-26 September, Beijing, China Aristotle University of Thessaloniki (AUTH)

  15. PCR product obtained from the amplification of mitochondrial 16S rRNA PCR successful even with CYSTS Artemia cysts Artemia adult INCO meeting, 23-26 September, Beijing, China Aristotle University of Thessaloniki (AUTH)

  16. Discriminant function analysis based on morphometry of bisexual Artemia populations INCO meeting, 23-26 September, Beijing, China Aristotle University of Thessaloniki (AUTH)

  17. MAD Discriminant function analysis based on morphometry of parthenogenetic Artemia populations (clade A) INCO meeting, 23-26 September, Beijing, China Aristotle University of Thessaloniki (AUTH)

  18. MAD Discriminant function analysis based on morphometry of parthenogenetic Artemia populations INCO meeting, 23-26 September, Beijing, China Aristotle University of Thessaloniki (AUTH)

  19. Contributors • INCO 3: AUTH, Greece • A.D. Baxevanis • A. Tzika • E. Markatzinou • A.C. Triantafyllidis • T.J. Abatzopoulos • INCO 14: ULA, Chile • INCO 9: RU, South Africa • J. Crespo • G. Gajardo • H. Kaiser • T. Hecht INCO meeting, 23-26 September, Beijing, China Aristotle University of Thessaloniki (AUTH)

  20. Mediterranean Basin Regional Action • INCO 3: AUTH, Greece, A.D. Baxevanis, A.C. Triantafyllidis, T.J. Abatzopoulos (Moderator) • INCO 4: CSIC, Spain, F. Amat • INCO 5: La Sapienza, Italy, G. Mura • INCO 8: INAT, Tunisia, M. Romdhane (Additional contribution from ARC) INCO meeting, 23-26 September, Beijing, China Aristotle University of Thessaloniki (AUTH)

  21. Mediterranean Basin Regional Action • Description of the present status • Information on samples existing in cyst banks • Cyst hatchability • Sampling data, storage conditions, hatching characteristics • NEW sampling • Report on 2 new Greek parthenogenetic populations • Characterization of old and new Artemia populations • Use of multidisciplinary approaches (A. salina, Greek parthenogens, Egyptian populations) • Tracking down invasive species INCO meeting, 23-26 September, Beijing, China Aristotle University of Thessaloniki (AUTH)

  22. Mediterranean Basin Total number of samples: 384 INCO meeting, 23-26 September, Beijing, China Aristotle University of Thessaloniki (AUTH)

  23. Mediterranean Basin Geographical distribution of the recorded samples INCO meeting, 23-26 September, Beijing, China Aristotle University of Thessaloniki (AUTH)

  24. 1 18 48 137 25 1 15 27 16 6 10 42 Sites not shown Morocco: 1 Portugal: 31 Unknown: 5 Mediterranean Basin (Number of samples per country) INCO meeting, 23-26 September, Beijing, China Aristotle University of Thessaloniki (AUTH)

  25. Mediterranean Basin 1. Report of A. fransciscana in Mediterranean Basin (El Estanquillo saltern, Spain)  Invasive species? 2. 224 samples need identification 3. Coexistence of A. salina and parthenogenetic strains INCO meeting, 23-26 September, Beijing, China Aristotle University of Thessaloniki (AUTH)

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