Rosidiani, ErtaPuri (2012) Analisis Variasi Genetik Amorphophallus muelleri Blume dari Berbagai Populasi di Jawa Timur Berdasarkan Sekuen Intron trnL. Magister thesis, Universitas Brawijaya.
Abstract
Amorphophallus muelleri ditemukan dari Thailand tengah ke selatan melalui Sumatera, Jawa ke Kepulauan Sunda Kecil. Porang (A. muelleri Blume) adalah satu dari 27 jenis Amorphophallus di Indonesia dan dari 170 jenis yang dikenal di dunia. Penelitian ini bertujuan untuk mengetahui variasi genetik dan kekerabatan pada porang beda populasi berdasarkan analisis sekuen intron trnL. Penelitian ini dilakukan pada delapan populasi A. muelleri di enam Kabupaten di Jawa Timur, yaitu populasi Kabupaten Jember (Desa Mayang), Malang (Kecamatan Lawang dan Kecamatan Kalipare), Blitar (Desa Brongkos), Nganjuk (KPH Tritik desa Bendoasri), Madiun (Desa Klangon, Saradan), Bojonegoro (Kecamatan Klino). Pemilihan lokasi penelitian yang berbeda-beda bertujuan untuk mengetahui bagaimana hubungan kekerabatan individu A. muelleri pada lokasi penelitian tersebut, dan pemilihan lokasi penelitian didasarkan pada hasil penelitian sebelumnya yang dilakukan oleh Azrianingsih dkk (2008); Harijati dkk (2008); Indriyani (2011) dan Mastuti dkk (2008). Eksplorasi yang dilakukan bertujuan untuk mengamati morfologi tanaman dan mengambil sampel daun muda sebanyak tiga lembar pada dua individu berbeda yang ditemukan pada masing-masing lokasi. Sampel daun yang telah didapatkan selanjutnya dilakukan proses ekstraksi (isolasi) DNA, amplifikasi intron trnL dengan menggunakan primer universal ‘c` dan ‘d` . Setelah proses amplifikasi, dilanjutkan proses purifikasi dan sekuensing. Hasil sekuensing yang berupa sekuen selanjutnya dilakukan analisis variasi genetik dan dibuat dendrogram dengan software MEGA 5, selanjutnya dideskripsikan dan dibahas. Sebagai outgrup dipilih A. margaritifer dan A. konkanensis . Pemilihan tersebut berdasarkan hasil penelitian Sedayu dkk (2010) menggunakan analisis Maximum Parsimony yang menunjukkan bahwa A. margaritifer dan A. konkanensis berada satu klad dengan A. muelleri yaitu Kontinental Asia I. Sekuen A. margaritifer dan A. konkanensis diperoleh dari genBank NCBI dengan kriteria sekuen trnL (trn-Leu) gene partial, chloroplast . Hasil eksplorasi pada delapan populasi (area jelajah) didapatkan masing-masing dua individu porang. Keenambelas individu tersebut memiliki beberapa karakter yang berbeda. Dari hasil pengamatan karakter morfologi diketahui karakter yang paling menonjol adalah corak tangkai. Keragaman corak tangkai yang telah diketahui pada penelitian ini yaitu, (1) totol putih lonjong, prismatik memanjang; (2) totol putih corak bulat telur; (3) totol putih corak bulat, garis putih panjang; (4) totol putih corak prismatik pendek; (5) totol putih corak prismatik pendek, garis putih panjang; (6) Totol putih corak prismatik panjang; (7) totol putih corak prismatik panjang, garis panjang putih; dan (8) garis panjang berwarna putih. Individu-individu porang yang ditemukan dapat dikelompokkan menjadi delapan kelompok berdasarkan persamaan karakter coraknya yaitu (1) MY1; (2) LW2; (3) BL1; (4) LW2, KP1, BL2, TR2 dan KN2; (5) TR1; (6) KP2, MD1 dan KL2; (7) MY2 dan MD2; dan (8) KN1 dan KL2 (Tabel 5.1.2). Berdasarkan kelompok tersebut diketahui bahwa porang dengan beragam corak telah tersebar ke beberapa daerah. Persebaran porang ke beberapa daerah disebabkan oleh adanya upaya pengembangan porang di Jawa Timur sebagai komoditi ekspor.
English Abstract
Amorphophallus muelleri is found from central Thailand to the south through Sumatra, Java, Lesser Sunda Islands. Porang ( A. muelleri Blume) is one of 27 species of Amorphophallus in Indonesia and of the 170 species known in the world. The aims of this study were to determine the genetic variation, morphological and relationship of porang at different populations based on analysis of trnL intron sequences. The research was conducted in Province of Jawa Timur, which is at Kabupaten Jember (Desa Mayang), Malang (Kecamatan Lawang and Kalipare), Blitar (Desa Brongkos), Nganjuk (KPH tritik Desa Bendoasri), Madiun (Desa Klangon, KPH Saradan), Bojonegoro (Kecamatan Klino). The aim of selecting various sites to study is to determine how A. muelleri individuals kinship at various sites, and site of study is based on the results of previous studies conducted by Azrianingsih et.al (2008); Harijati et.al (2008); Indriyani (2011) and Mastuti et.al (2008). Exploration was conducted in six Kabupaten (8 population). The aims were observing the morphology and take samples of young leaves as many as three sheets at two different individuals were found at each location. Leaf samples that had been obtained, then were extracted the DNA, and amplified the trnL intron using universal primers c and d. After the amplification, amplicon were purified and sequenced. The results of sequencing were analyzed the genetic variation and relationship based on Maximum Parsimony tree which is created by software MEGA5, hereinafter described and discussed. The out-group were selected based on research results Sedayu et al (2010) using the Maximum Parsimony analysis showed that A. margaritifer and A. konkanensis is one clad with A. muelleri is Continental Asia I. The sequence of A. margaritifer and A. konkanensis were obtained from NCBI GenBank, with criteria trnL sequences (trn-Leu) gene partial, chloroplast. Exploration results in the eight populations (cruising area) obtained two individual porang each population. The sixteen individual has a different character. Observations of morphological characters showing the most prominent feature is the stalk pattern. The diversity of patterns that have been known in this study, namely: (1) white spots oval, elongated prismatic; (2) white spotted an oval pattern; (3) circular pattern of white spots, a long white line; (4) short prismatic shades of white spots; (5) short prismatic shades of white spots, a long white line; (6) spotted long white prismatic shades; (7) long prismatic shades of white spots, a long line of white; (8) a long line of white. Porang individuals were found to be grouped into eight groups based on the similarity character of stalk pattern; (1) MY1; (2) LW2; (3) BL1; (4) LW2, KP1, BL2, TR2 and KN2; (5) TR1; (6) KP2, MD1 and KL2; (7) MY2 and MD2; and (8) KN1 and KL2. Porang with various shades have spread to some areas. Distribution porang to some areas due to efforts to development porang in East Java as an export commodity. Porang genetic diversity in East Java in this study were analyzed based on trnL intron sequences in each individual plant porang found. Sequencing of 16 individuals porang, showed 13 individuals which have a clear chart chromatography which is resulting in the proper sequence and can be used in subsequent analyzes. Three individual sequences that can not be used are LW1, KP2, and BL2, because the sequence has a low quality of chromatographic chart that can not be read clearly, so resulting ambiguous sequences in readings. Based on the sequences of 13 individual porang, showing the differences in nucleotide base composition. Six region is also known as conserved region, there are sequences: 54-151: 153-177: 179-285: 287-302: 337-430; and 432-458. In the analysis of genetic variation, known to the individuals who experience frequent changes in the nucleotide bases and deletion. The sequence of nucleotide base changes in individuals with the most frequent to rare is KP1, KN2, BL1 and TR2, LW2, MD2, KL2, KN1, MY1, TR1, MD1, M
| Item Type: | Thesis (Magister) |
|---|---|
| Identification Number: | TES/584.64/ROS/a/041204156 |
| Subjects: | 500 Natural sciences and mathematics > 584 Liliopsida (Monocotyledons) > 584.6 Cyclanthales, Arales, Pandanales, Typhales |
| Divisions: | S2/S3 > Magister Matematika, Fakultas MIPA |
| Depositing User: | Endro Setyobudi |
| Date Deposited: | 20 Feb 2013 14:25 |
| Last Modified: | 20 Feb 2013 14:25 |
| URI: | http://repository.ub.ac.id/id/eprint/157820 |
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