2230_I Made Budiarsa.pdf - Universitas Gadjah Mada

No. KontrakBidang IImuKlasterLPPM-UGM/1198/2009MIPASains dan TeknoloaiLAPORAN AKHIR KEGIATANHIBAH PENELITIAN UNTUK MAHASISWA PROGRAM DOKTORTAHUN ANGGARAN 2009Judul PenelitianKajian Molekular Burung Maleo (Macrocepha/onBerdasarkan Sekuen Gen ND2 Mitokondriama/eo)Peneliti:Drs. I Made Budiarsa, M.Si.FAKULTAS BIOLOGIUNIVERSITAS GADJAH MADA2009

RINGKASANBurung maleo (Macrocephalon maleo) merupakan burung endemik Sulawesi yangdiklasifikasikan sebagai Endengared Species (IUCN) sehingga tindakan pelestarian perludilakukan. Salah satu aspek penting dan dapat digunakan sebagai dasar tindakan pelestarianadalah menggali seluruh informasi yang ada khususnya diversitas genetik dan hubungankekerabatan maleo. Tujuan penelitian ini adalah mengungkap diversitas genetik maleo,hubungan kekerabatan antar populasi maleo berdasar pada sekuen gen ND2 mitokondria.DNA darah diisolasi dan dipakai cetakan untuk amplifikasi gen ND2 dengan teknik PCR(Polymerase Chain Reaction). Produk PCR selanjutnya disekuensing kemudian digunakansebagai dasar untuk mengungkap diversitas genetik maleo dan hubungan kekerabatannya.Multiple alignments dilakukan dengan Program Clustal-X, identifikasi haplotipe dengan MEGAversi 2.1 dan nilai diversitas nukleotida digunakan Program DnaSP Versi 3.51. Konstruksiphylogenetic ,tree menggunakan Program Phylip dengan algorithm Neighbor Joining danmodel evolusi Juke-Cantor. Nilai similaritas dan jumlah nukleotida yang berbeda dianalisisdengan program Phydith.Hasil analisis, 1072 bp fragmen ND2 menghasilkan komposisi basa 10.1% G, 37.2%C, 22.7% T dan 30.0% A serta nilai diversitas genetik antara 0,011-0,020. Untuk seluruhsampel yang disertakan dalam analisis ini teridentifikasi 523 (48.8%) daerah variabel dan 418(39%) merupakan daerah informatif dengan komposisi bass 10,2% G, 36,6% C, 23,5% Tdan 29,7% A . Analisis filogenetik menunjukkan maleo adalah monofili dari genusMacrocephalon yang terdiri dari 3 clade terpisah dan didukung oleh nilai bootstrap >50 dannilai similaritas antara 97,69-99,63% serta berkerabat dekat dengan Aepypodius, Talegalla,Leipoa and Alectura1

SummaryThe maleo (Macrocepha/on ma/eo) is an endemic bird of Sulawesi, classified as anEndangered Species on the International Union for Conservation of Natural Resources(IUCN). One of the most important methods to secure the bird is to examine its geneticdiversity, relationship among populations, which is a fundamental component for conservingmaleo. The aim of this research is to examine the genetic diversity, relationship populations ofmaleo based on ND2 mitochondrial gene sequences, to implement it as a consideration torecommend conservation action of maleo, as well as for other Megapodes.Blood DNA was extracted and used as template for ND2 amplification using PCR(Polymerase Chain Reaction) method. PCR product was sequenced and used to examine thegenetic diversity and phylogeny of maleo. Multiple sequences alignment was obtained usingthe Clustal-X. Initial sequences comparison and iqentification of haplotypes were performedusing MEGA version 2.1. The values of nucleotides diversity were carried out by DnaSPversion 3.51. A phylogenetic tree was constructed using Phylip with Neighbor-JoiningAlgorithm and an evolution model Juke-Cantor. The similarity value and the nucleotidedifferences were analyzed using Phydith.Result indicated that from 1072 bp of ND2 sequence, the base composition included10.1% G, 37.2% C, 22.7% T and 30.0% A. Nucleotide diversity of maleo was between 0,011-0,020. All species analyzed in this research showed, 523 (48%) variable region and 418(39%) phylogeneticaly informative region and the base composition of ND2 fragment asfollow 10,2% G, 36,6% C, 23,5% T and 29,7% A. The phylogenetic analysis showed thatmaleo was monophyly of Macrocepha/on and showed 3 separated clades with bootstrapsupport >50 and similarity value was 97.69 - 99.63%. Maleo was closely related toAepypodius, Ta/egalla, Leipoa and A/ectura .11