Phylogenetic relationships of cockatoos (Aves: Psittaciformes) based on DNA sequences of the seventh intron of nuclear β-fibrinogen gene Dwi Astuti Zoological Division, R.C. for Biology - Indonesian Institute of Sciences, Cibinong Science Centre Indonesia [email protected] O Cockatoos are belonged to Cacatuinae (Forshaw, 1989) order : Psittaciformes, Cockatoo Distribution family : Psittacidae (Forshaw, 1989) Calopsittacini Chalyptorhynchini Cacatuini Cacatuidae (del Hoyo,1998) Nymphicus subfamily: Cacatuinae Probosciger Calyptorhynchus Challocephalon Eolophus Cacatua tribes : Calopsittacini Chalyptorhynchini Cacatuini E. roseicapillus Probosciger N. hollandicus P. aterrimus C. baudinii C. fimbriatum C. sulphurea O In the world, there are six extant genera C. latirostris C.galerita Eolophus C. lathami C. alba Cacatua consisting of 21 cockatoo species C. banksii Nymphicus C. moluccensis leadbeateri C. funereus C. goffini Calyptorhynchus O Some previous authors have made grouping C. magnificus C. sanguinea Calyptorhynchus Callocephalon C. leadbeateri and evolutionary relationships of cockatoos C. ophthalmia Brown & Toft (1999) based on morphological characters, isozyme, C. haematuropygia and mitochondrial DNA. However, their MATERIALS AND METHODS relationships are still controversial, especially Phylogeny based on different characters concerning the position of Nymphycus Cacatua leadbeateri Blood samples from each individual of 15 species, Biochemical 6 genera, and 3 tribes of cockatoos hollandicus. Since the nuclear β-fibrinogen (Adams et al. 1984) Cacatua galerita gene has been recognized useful for Cacatua sanguinea phylogenetic studies of some birds, this study Cacatua tenuirostris preserved in 99% ethanol employed the DNA sequence of seventh intron Eolophus roseicapillus of this gene (fibint7) to construct phylogenetic Callocephalon DNA extraction (from 5 – 20 mg of dry blood) relationships of cockatoos based on neighbor- Nymphicus joining (NJ) and maximum-parsimony (MP) Calyptorhynchus Fibint7 Amplification (PCR) methods Morphology Cacatua Other parrots (del Hoyo et al., 1997) Eolophus Cacatua DNA sequencing Callocephalon Cacatua leadbeateri Unlike previous studies, DNA Nymphicus Eolophus DNA sequence data analyses (DNAsys, ProSeq, sequence data from nuclear β- Calyptorhynchus Callocephalon Probosciger MEGA,PAUP) fibrinogen gene (Fibint7) of six Nymphicus All other parrots Calyptorhynchus cockatoo genera were analyzed in Origin of Constructing the phylogenetic trees : MP Probosciger this study parrots Mt 12S (Brown & (maximum parsimony) & NJ (neighbor- Toft, 1999) Objectives of this study outgroup joining) 1. To resolve the phylogenetic relationships within cockatoos Sequence Characteristics of fibint7 on Cockatoos (Cacatuinae phylogeny) Saturation analyses

30 40 2. To determine the position of cockatiel Length of indels : 1 – 9 bp 35 25 (Nymphicus) 30 20 25 3. To determine the monophyly or No. of nucleotide : 808 – 817 bp 15 20 15

Transition 10 paraphyly of each tribe Transversion 10 Variable sites : 310 5 5 4. To determine whether or not the 0 0 0 0.02 0.04 0.06 0.08 0 10203040 phylogeny inferred from Fibint7 Transversion Distance reflects biogeography of the Informative sites : 164 cockatoos Phylogenetic trees of cockatoos MP based on fibint7 of β-fibrinogen gene

NJ tree MP tree CONCLUSION Cacatua sulphurea citrinocristata Cacatua sulphurea citrinocristata 64 Cacatua s. sulphurea Cacatua s. sulphurea 64 Cacatua galerita eleonora Cacatua galerita eleonora Թ Relationships between species of genus Cacatua were Cacatua g. triton N.G & Cacatua g. triton not well resolved by Fibint7 gene sequence. 84 Cacatua g. galerita Indonesia Cacatua g. galerita 57 Cacatua alba Cacatua alba Թ Cacatua leadbeateri was relatively distant from Cacatua moluccensis Cacatua moluccensis 71 Cacatua goffini Cacatua goffini 84 60 congeneric species Cacatua sanguinea 91 Cacatua sanguinea 67 Cacatua leadbeateri Australia Tribe Cacatuini (Cacatua, Eolophus and Cacatua leadbeateri 55 Թ Eolophus roseicapillus Eolophus roseicapillus NG, Indonesia, Callocephalon) was monophyletic 72 Callocephalon fimbriatum Callocephalon fimbriatum Australia Probosciger aterrimus Probosciger aterrimus Թ Tribe Calyptorhynchini (Calyptorhynchus and Calyptorhynchus banksii Calyptorhynchus banksii 98 99 94 Probosciger ) was paraphyletic 91 Calyptorhynchus lathami Australia Calyptorhynchus lathami 69 89 98 Calyptorhynnchus baudinii 100 Calyptorhynnchus baudinii Թ Nymphicus probably a basal clade of cockatoos Calyptorhynchus latirostris Calyptorhynchus latirostris 96 Nymphycus hollandicus Nymphycus hollandicus Թ Phylogeny inferred from Fibint7 sequences reflects Psittaculirostris Psittaculirostris biogeography of cockatoos. Probably, cockatoos Columba Columba came from Australia and spread to Pacific region, Accipiter Accipiter including N.G and Indonesian Archipelago Calopsittacini Chalyptorhynchini Cacatuini

ACKNOWLEDGEMENT The research was supported by Centre of Excellent Project, Hokkaido University, and Japan Society for Promoting Science (JSPS) Core University (Hokkaido University and LIPI). I would like to thank: 1) Prof. Seigo Higashi and Dr. Hitoshi Suzuki (Hokkaido University) for kindly supervising me; and 2) Dr. Ken Aplin (CSIRO) for sharing the samples