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Barcoding Neotropical Psittacidae

Barcoding Neotropical Psittacidae. ABBI Regional Working Group Meeting Buenos Aires, 14-16 March 2007 Cristina Y. Miyaki Departamento de Genética e Biologia Evolutiva Instituto de Biociências Universidade de São Paulo. Notropical Psittacidae or tribe Arini (Smith 1975).

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Barcoding Neotropical Psittacidae

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  1. Barcoding Neotropical Psittacidae ABBI Regional Working Group Meeting Buenos Aires, 14-16 March 2007 Cristina Y. Miyaki Departamento de Genética e Biologia Evolutiva Instituto de Biociências Universidade de São Paulo

  2. Notropical Psittacidae or tribe Arini (Smith 1975) 150 species [total in the Order: 330] (del Hoyo et al. 1997) 30 genera [total in the Order: 84] (del Hoyo et al. 1997) Psittaciformes: distribution

  3. Nestor notabilis Amazona farinosa 100 Graydidascalus brachyurus Pionus maximiliani Anodorhynchus hyacinthinus Ara ararauna Orthopsittaca manilata 100 Primolius auricollis 99 Nandauys nenday 100 Diopsittaca nobilis 100 Guarouba guarouba Enicognathus leptorhynchus Rhynchopsitta pachyrhyncha 100 Hapalopsittaca amazonina Pionites melanocephala 100 Bolborhynchus lineola Nannopsittaca panychlora 99 Myiopsitta monachus Forpus crassirostris Chalcopsitta cardinalis 100 Pseudeos fuscata 100 Trichoglossus haematodus 100 Lorius garrulus 99 Melopsittacus undulatus Neophema elegans 98 Agapornis roseicollis 100 Eclectus roratus Psittacula krameri 100 Poicephalus senegalus Psittacus erithacus Cacatua galerita Monophyly of Arini Bayesian unrooted (3062 bp 12S, 16S &intron 5 tropomyosin) Red = Neotropical Blue = African Green = Australasian Bold: no MP support, only Bayesian Matsumoto et al, prep.

  4. Photo: P.J. Faria Arini 150 species 27% endangered Source: Forshaw 1989

  5. Recife airport: to Lisbon 58 eggs (possibly parrots) Incubator: non ideal conditions Visited many places in Brazil (Amazon, Mato Grosso, Tocantins and Brasília) Photo: Folha on line Application of DNA barcodes in Neotropical Psittacidae Species identification of non hatched embryos Which species?

  6. Species identification of non hatched embryos Choice of marker: mitochondrial 16S rDNA Sequenced 141-509 bp: 6 groups (haplotypes) Analyses: BLAST (GenBank) and distance Matsumoto, Oliveira-Marques, et al., in prep.

  7. Salvatoria xanthops (50) Amazona aestiva/ Amazona ochrocephala (4) 213 bp of 16S rDNA Ara ararauna (3) Non psitacid(1) Matsumoto, Oliveira-Marques, et al., in prep.

  8. Ara macao Ara chloroptera Ara militaris Ara ambigua Primolius maracana Primolius auricollis Primolius couloni Guarouba guarouba Diopsittaca nobilis Orthopsittaca manilata Ara ararauna Group 2 Ara glaucogularis Aratinga cactorum Ara severa Cyanopsitta spixii Anodorhynchus hyacinthinus 390 bp of 16S rDNA Ara ararauna (3)

  9. Species identification of non hatched embryosConclusions The available 16S rDNA data base was informative to identify the species of the embryos. The species identification was confirmed in another lab that used another mitochondrial marker (cytochrome b). Would COI be a good marker? Building a COI database for Neotropical parrots and will test the same embryos.

  10. COI sequence database for Neotropical parrots Master’s Degree student: Priscila F.M. Gonçalves (Dec 2006) Objective: construct the database and evaluate its usefulness for species identification (e.g. evaluate if genetic distances within and among taxa do not overlap)

  11. Primers LTyr (O. Haddrath) Bird R1 (Hebert et al 2004) COI H7557 (P. Gonçalves) COI H7557 LTyr Bird R1 826 bp 980 bp COI sequence database for Neotropical parrots • Samples 113 out of 150 species of Neotropical parrots (34 with skins)

  12. COI sequence database for Neotropical parrots • PCR conditions (touchdown) 5 min 95ºC; 14 cycles: 30 sec 95ºC, 30 sec 68ºC (1°C lower per cycle), 40 sec 72ºC; 29 cycles: 30 sec 95°, 30 sec 54°C, 40 sec 72°C; 10 min 72°C • Sequences obtained: 23 (March 2) • Average size of sequences: 792 bp (732-853 bp)

  13. Acknowledgements Erika Tavares, Royal Ontario Museum, Toronto

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