Barcoding Neotropical Psittacidae

ABBI Regional Working Group Meeting

Buenos Aires, 14-16 March 2007

Cristina Y. MiyakiDepartamento de Genética e Biologia

EvolutivaInstituto de Biociências

Universidade de São Paulo

Psittaciformes: distribution

150 species [total in the Order: 330] (del Hoyo et al. 1997)

30 genera [total in the Order: 84] (del Hoyo et al. 1997)

Notropical Psittacidae or tribe Arini (Smith 1975)

Monophyly of AriniBayesian unrooted (3062 bp 12S, 16S & intron 5 tropomyosin)

Red = NeotropicalBlue = AfricanGreen = AustralasianBold: no MP support, only Bayesian

Nestor notabilisAmazona farinosaGraydidascalus brachyurusPionus maximilianiAnodorhynchus hyacinthinusAra araraunaOrthopsittaca manilataPrimolius auricollisNandauys nendayDiopsittaca nobilisGuarouba guaroubaEnicognathus leptorhynchusRhynchopsitta pachyrhynchaHapalopsittaca amazoninaPionites melanocephalaBolborhynchus lineolaNannopsittaca panychloraMyiopsitta monachusForpus crassirostrisChalcopsitta cardinalisPseudeos fuscataTrichoglossus haematodusLorius garrulusMelopsittacus undulatusNeophema elegansAgapornis roseicollisEclectus roratusPsittacula krameriPoicephalus senegalusPsittacus erithacusCacatua galerita

99

99

100

100

100

99

100

100

100

98

100100

100

100

100

Matsumoto et al, prep.

Source: Forshaw 1989

Arini

150 species

27% endangered

Photo: P.J. Faria

Species identification of non hatched embryos

Application of DNA barcodes in Neotropical Psittacidae

Which species?

Photo: Folha on line

Recife airport: to Lisbon

58 eggs (possibly parrots)

Incubator: non ideal conditions

Visited many places in Brazil (Amazon, Mato Grosso, Tocantins and Brasília)

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.

Species identification of non hatched embryos

Matsumoto, Oliveira-Marques, et al., in prep.

213 bp of 16S rDNA

Ara ararauna (3)

Salvatoria xanthops (50)

Amazona aestiva/ Amazona ochrocephala

(4)

Non psitacid (1)

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 cactorumAra severa

Cyanopsitta spixii

Anodorhynchus hyacinthinus

390 bp of 16S rDNA

Ara ararauna

(3)

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).

Species identification of non hatched embryos

Conclusions

Would COI be a good marker?

Building a COI database for Neotropical parrots and will test the same embryos.

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)

•Samples

113 out of 150 species of Neotropical parrots (34

with skins)

COI sequence database for Neotropical parrots

826 bp

980 bp

LTyr Bird R1 COI H7557

•Primers

LTyr (O. Haddrath)

Bird R1 (Hebert et al 2004)

COI H7557 (P. Gonçalves)

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)

Acknowledgements

Erika Tavares, Royal Ontario Museum, Toronto