HUBUNGAN FILOGENETIK DUA POPULASI KATAK POHON infrafrenatus infrafrenatus (TYLER 1971) DI INDONESIA BERDASARKAN GEN 16S rRNA

Taufan Nurzaman Sulaeman (G352180071)

Dr. Ir. Achmad Farajallah, MSi. Dr. Amir Hamidy, MSc.

Sekolah Pascasarjana ZOOGEOGRAPHY (BIO 562) Institut Pertanian Bogor 2020 GRADUATE SCHOOL IPB UNIVERSITY Nyctimystes infrafrenatus (Gunther 1867) Katak Pohon Hijau Papua | White Lipped Treefrog| Australian Giant Treefrog

Dorsal berwarna hijau

Dorsal berwarna putih

Bibir bawah (infralabial) berwarna putih. Strip putih memanjang hingga ke bagian bawah tympanum

Jari kaki depan Jari kaki belakang berselaput sebagian Berselaput penuh

SVL = 130 mm (Menzies 2006) 2 Latar Belakang Permasalahan Sistematik : Morfologi : Molekuler : Nyctimystes (Menzies 2006; Pyron dan Wiens 2011; Duellman et al. 2016)

Litoria

Nyctimystes

Ranoidea

Duellman et al. 2016) Pyron dan Wiens 2011 3 Latar Belakang Distribusi luas : Wilayah timur Indonesia, Papua Nugini utara , Kaledonia Baru dan Kepulauan Solomon (Iskandar dan Colijn 2000; Menzies 2006; Iskandar et al. 2017; Frost 2020)

Species Kriptik (?) Kepulauan Maluku : Chalcorana chalconota (Inger et al. 2009) Kei, Ambon, Obi, Buru, Seram, Rhacophorus dugritei (Li et al. 2012) Talaud, Morotai, Halmahera Polypedates leucomystax (Kuraishi et al. 2013) Leptobrachium hasseltii Papua : (Hamidy dan Matsui 2017) Papua, Papua Barat, Aru, Waigeo, Microhyla complex (Atmadja et al. 2018). Misol, Tanimbar

(Brongersma 1948; Boulenger 1912; Tyler 1978; Setiadi dan Hamidy 2006; Hamidy dan Mulyadi 2007; Koch et al. 2007; Karin et al. 2018; Nugroho et al. 2019).

Peta distribusi Nyctimystes infrafrenatus

4 Latar Belakang 2 Subspesies: Nyctimystes infrafrenatus infrafrenatus Nyctimystes infrafrenatus militarius (Tyler 1971)

Subspesies Nyctimystes infrafrenatus militarius Sinonim Hyla militaria Subspesies : Nyctimystes infrafrenatus infrafrenatus Hyla dolichopsi var. pollicaris

Sinonim : Calamites dolichopsis Litoria guttata Hyla dolichopsi var. tennuigranulata Hyla spengeli) (Loveridge 1948; Tyler 1971; Iskandar dan Colijn 2000; Menzies 2006).

5 Nyctimystes infrafrenatus infrafrenatus Latar Belakang 2 populasi alopatrik - 2 subregion

Nyctimystes infrafrenatus infrafrenatus Nyctimystes infrafrenatus infrafrenatus Populasi Maluku Populasi Papua Wallacea Australo-Papua

Molekuler ???: Morfometrik: Perbedaan pada Gen 16S rRNA ukuran tungkai (Vences et al. 2005) belakang betina (Prafiadi et al. 2016)

• Sejarah geologi pembentukan kawasan yang berbeda (Hall 1998; Hall 2002; Hall 2009; Hall 2013). • Amfibi endemik terhadap benua atau unit biogeografi tunggal (Vences dan Kohler 2008 • Distribusi amfibi melalui laut terbatas (Duellman dan Trueb 1986; Vences et al. 2003; Fonte et al. 2019) 6 Latar Belakang

 Permasalahan Sistematik  Distribusi luas  Dua populasi pada dua subregion biogeografi  Perbedaan morfometrik*

7 Tujuan

• Menentukan status taksonomi N. i. infrafrenatus • Mengevaluasi hubungan kekerabatan antara populasi N. i. infrafrenatus Papua dan Maluku

8 Metode Penelitian Waktu dan Tempat • November 2019 – April 2020 • Laboratorium Herpetologi, Lembaga Ilmu Penelitian Indonesia (LIPI), Cibinong, Bogor.

Sampel Penelitian

• Sediaan sekuens gen 16S rRNA Nyctimystes infrafrenatus infrafrenatus Laboratorium Herpetologi, LIPI Primer 16S rRNA : 16 H-1 (5’- CTCCGGTCTGAACTCAGATCACGTAGG -3’) 16 L-1 (5’- CTGACCGTGCAAAGGTAGCGTAATCACT –3’)

• Sediaan Sekuens 16S rRNA Nyctimystes infrafrenatus infrafrenatus dan outgrup yang tersedia di Genbank (https://www.ncbi.nlm.nih.gov/genbank/)

9 METODE Metode Penelitian

Pensejajaran Sekuens Rekonstruksi Pohon Filogenetik Visualisasi

GTR + G 1000 replikasi Maximum Likelihood

Sekuens LIPI (ML) Perangkat lunak : MEGA X

Hasil ALIGNMENT Pensejajaran MUSCLE Jarak genetik (p-distance) Perangkat lunak : 419 pb MEGA X Sekuens Genbank Perangkat lunak : MrBayes 3.2.7a Bayesian Inference Perangkat lunak : (BI) FigTree GTR + G 10.000.000 MCMC Frekuensi diagnosis 100.000 Burn-in 25 % Topologi sampel tiap 1000 generasi

1010 Metode Penelitian

Table 1. Sampel N.i. infrafrenatus dan outgroup yang digunakan pada rekonstruksi pohon filogenetik.

1111 METODE Metode Penelitian

Table 1 lanjutan.....

1212 Hasil dan Pembahasan A

• Monofiletik • Membentuk 2 subclade B

Gambar 1. A) Spesimen N. i. infrafrenatus dari Timika, Papua memiliki “bibir putih” yang lebih tebal dari N. i. infrafrenatus dari Ternate, Maluku. Foto : Amir Hamidy (A) dan M. Iqbal Setiadi (B).

Gambar 2. Pohon filogenetik berdasarkan topologi Bayesian Inference N. i. infrafrenatusdan outgrup berdasarkan sekuens 16Sr RNA (419 pb). Angka pada lengan pohon menunjukkan besarnya bootstrap ML (MLBS) dan probabilitas posterior (BPP). 13 Hasil dan Pembahasan Table 2. Jarak genetik (uncorrected p-distances) diantara populasi N. i. infrafrenatus dan spesies lain dari genus Nycimystes

• Jarak genetik 3% , indikasi pemisahan tingkat species (Fouquet et al. 2007) • Leptobrachium hasseltii Bali dan Jawa 3-4% (Hamidy dan Matsui 2017)

Ranoidea bella sp. nov dan Ranoidea auae 2.00 % (McDonald et al. 2016)

Gambar 2. Pohon filogenetik berdasarkan topologi Bayesian Inference N. i. infrafrenatusdan outgrup berdasarkan sekuens 16Sr RNA (419 pb). Angka pada lengan pohon menunjukkan besarnya bootstrap ML (MLBS) dan probabilitas posterior (BPP). 14 Hasil dan Pembahasan Vicariance memisahkan Papua dari Australia (Hantoro et al. 1995; Van der Kaars 1991; Voris 2000) 20 mya 15 mya P-distance : ] 1.3-1.5%

(18,000 14C yr BP) (16,000 14C yr BP) 10 mya 5 mya

(12,000 14C yr BP) (9000 14C yr BP) (6000 14C yr BP)

Perubahan permukaan air laut dan garis pantai pada masa glasiasi maksimum terakhir (LGM) memunculkan dangkalan Sahul (Yokoyama et al. 2001) 15 Hasil dan Pembahasan Vicariance memisahkan Papua dari Australia (Hantoro et al. 1995; Van der Kaars 1991; Voris 2000) 20 mya 15 myaJembatan distribusi/pertukaran Spesies • hutan tropis dataran rendah • padang rumput-semak • sistem sungai (Hantoro et al. 1995; Van der Kaars 1991; Voris 2000)

(18,000 14C yr BP) (16,000 14C yr BP) 10 mya 5 mya

(12,000 14C yr BP) (9000 14C yr BP) (6000 14C yr BP)

Perubahan permukaan air laut dan garis pantai pada masa glasiasi maksimum terakhir (LGM) memunculkan dangkalan Sahul (Yokoyama et al. 2001) 16 Hasil dan Pembahasan Halmahera adalah Oceanic Islands (Hall 1998; Hall 2002; Hall 2009; Lohman et al. 2011; Hall 2013).

20 mya 15 mya

Mekanisme distribusi spesies pada oceanic island : 10 mya 5 mya overseas/overwater jump dispersal”

Katak : Boophis dan Mantidactylus Madagascar  Comoros (Vences et al. 2003; Fonte et al. 2019).

Tropicagama temporalis (Kadal) Kolonisasi Maluku dari Australia Gambar Palaeogeografi Wallacea (Hall 2013) melalui Papua (Karin et al. 2020) 17 Simpulan

• Populasi N. i. infrafrenatus di Indonesia merupakan Clade monofiletik dengan dua subclade • Berdasarkan pemisahan subclade pada pohon filogenetik dan jarak genetik diatas ambang batas spesies, populasi N. i. infrafrenatus Maluku merupakan spesies yang berbeda dari populasi Papua. • Secara morfologi terdapat perbedaan tebal garis putih pada infralabial yang dapat menjadi salah satu karakter diagnostik untuk pendeskripsian spesies lebih lanjut.

18 Terima Kasih

1919 Daftar Pustaka

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