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I Made Tasma
Indonesian Center for Agricultural Biotechnology and Genetic Resources Research and Development, Jl. Tentara Pelajar 3A, Bogor 16111, Indonesia Phone (+62-251) 8337975
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Decision Sciences, Operations Research & Management Industrial & Manufacturing Engineering

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Phylogenetic and Maturity Analyses of Sixty Soybean Genotypes Used for DNA Marker Development of Early Maturity Quantitative Trait Loci in Soybean Tasma, I Made; Satyawan, Dani; Warsun, Ahmad; Yunus, Muhamad; Santosa, Budi
Jurnal AgroBiogen Vol 7, No 1 (2011): Jurnal AgroBiogen
Publisher : Jurnal AgroBiogen

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The Indonesian soybean productivity is still very low with the national average of 1.3 t/ha. One means to improve national soybean productivity is by manipulating harvest index by cultivating very early maturing soybean cultivars. Development of early maturing soybean cultivars can be expedited by using marker-aided selection. The objective of this study was to select parental lines having contrasted maturity traits and selected parents must be genetically distance. The parents then were used to develop F2 populations for detecting early maturity QTL in soybean. Maturity tests of 60 soybean genotypes were conducted at two locations, Cikeumeuh (Bogor) and Pacet (Cianjur) using a randomized block design with three replications. Genomic DNA of the 60 genotypes were analyzed using 18 SSR markers and genetic relationship was constructed using the Unweighted Pair-Group Method Arithmatic through Numerical Taxonomy and Multivariate System program version 2.1-pc. Results showed that the 60 genotypes demonstrated normal distribution in both locations for days to R1 (32-48d), days to R3 (35-55d), days to R7 (75-92d), and days to R8 (78-99d). Four early maturing genotypes and three late genotypes were obtained. Total SSR alleles observed were 237 with average allele per locus of 12.6 (3-29), and average PIC value of 0.78 (0.55-0.89). Genetic similarity among genotypes ranges from 74.8-95%. At similarity level 77% divided the genotypes into six clusters (the four selected early maturing genotypes located in clusters III and IV, while the three late genotypes located in cluster II). Based on maturity data, pubescent color, and phygenetic analysis seven parents were selected (four early maturing genotypes B1430, B2973, B3611, B4433 and three late genotypes B1635, B1658, and B3570). Twelve F2 populations were developed with the aid of SSR markers Satt300 dan Satt516. Two of the populations will be used to develop DNA markers for earliness in soybean.
Konstruksi Pustaka Genom Kakao (Theobroma cacao L.) untuk Sekuensing Genom Total Menggunakan Next Generation Sequencing HiSeq2000 Tasma, I Made; Satyawan, Dani; Rijzaani, Habib; Rubiyo, Rubiyo
Jurnal Tanaman Industri dan Penyegar Vol 3, No 2 (2012): Buletin Riset Tanaman Rempah Dan Aneka Tanaman Industri
Publisher : Pusat Penelitian dan Pengembangan Perkebunan

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Abstract

Pemuliaan kakao secara konvensional memerlukan waktu panjang (10-15 tahun). Pemanfaatan marka DNA akan memperpendek siklus pemuliaan kakao. Tujuan penelitian ini adalah mengkonstruksi pustaka genom tiga genotipe kakao yang dapat digunakan untuk sekuensing genom total kakao menggunakan NGS HiSeq2000 dan mendapatkan data resekuen genom total tiga genotipe kakao.  Bahan tanaman terdiri dari tiga klon unggul kakao (ICCR02, ICCR04, dan SUL02) diperoleh dari Balittri, Pakuwon.  DNA genomik diisolasi dari daun muda sebagai bahan konstruksi pustaka genom total. Sekuensing pustaka dilakukan pada mesin HiSeq2000 mengikuti protokol dari Illumina. Pustaka genom yang telah berhasil dikonstruksi berukuran 300 pasang basa (bp) masing-masing dengan konsentrasi 14,70 ng/µL (ICCR02), 15,20 ng/µL (ICCR04), dan 12,90 ng/µL (SUL02). Ukuran dan konsentrasi pustaka genom yang dihasilkan sangat ideal untuk sekuensing menggunakan HiSeq2000. Sekuensing ketiga genom menghasilkan data sekuen 52,9 x 109 bp.  Klaster DNA pustaka genom memiliki nilai Q scores>30 (75,0%) dengan tingkat kesalahan pembacaan basa rendah (1,47%).  Nilai densitas klaster, persen klaster PF, intensitas basa, persen phasing, dan persen prephasing menunjukkan kualitas klaster pustaka genom ketiga genotipe kakao termasuk kategori pustaka ideal. Data sekuen yang dihasilkan juga sangat ideal untuk identifikasi marka SNP genom kakao. Koleksi marka SNP digunakan untuk identifikasi gen pengendali karakter penting kakao dan pemuliaan berbasis marka DNA untuk memperpendek siklus pemuliaan kakao. Genomic Library Construction Of Cocoa (Theobroma Cacao L.) For Whole Genome Sequensing Using A Next Generation Sequencer Hiseq2000Conventional cocoa breeding is slow and takes about 10-15 years to complete a breeding cycle. Applying genomic technology using DNA markers will significantly decrease cocoa breeding cycle. The objectives of this study were to construct cocoa whole genome genomic libraries to be used for resequencing the whole genome of cocoa and obtain whole genome resequence data of three cocoa genotypes. Three Indonesian cocoa genotypes (ICCR02, ICRR04, and SUL02) were used. DNA genomic was isolated from young leaf and used to construct genomic DNA libraries and generate DNA clusters. DNA clusters were sequenced using a HiSeq2000 platform. The whole genome libraries of the cocoa genotypes were successfully constructed. The library size was 300 bp with concentrations of 14.70 ng/µL (ICCR02), 15.20 ng/µL (ICCR04), and 12.90 ng/µL (SUL02), respectively. The genomic library size and concentrations are suitable for sequencing study using the NGS HiSeq2000. Total sequencing output obtained was 52.9 x 109 bp. The genomic library clusters resulted during the sequencing process demonstrated the Q scores > 30 of 75.0% with low error sequencing rate of 1.47%. Cluster densities, percentage of cluster PF, base intensity, and percentage of phasing and prephasing indicated the cluster quality of the genomic libraries is classified as an ideal one to be used for resequencing study using NGS HiSeq2000. The resequence data were ideal for SNP marker discovery. SNP markers are used to identify economically important genes of cocoa and marker-aided cocoa breeding to decrase the cocoa breeding cycle.
ANALISIS DIVERSITAS GENETIK AKSESI KELAPA SAWIT KAMERUN BERDASARKAN MARKA SSR TASMA, I MADE; ARUMSARI, SEKAR
853-8212
Publisher : Pusat Penelitian dan Pengembangan Perkebunan

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ABSTRAKDiversitas genetik aksesi kelapa sawit Indonesia saat ini sangatrendah. Dalam usaha meningkatkan keragaman genetik telah dilakukaneksplorasi plasma nutfah di pusat keragaman genetik kelapa sawit diKamerun. Tujuan dari penelitian ini untuk mengetahui diversitas genetikdan tingkat polimorfisme berdasarkan marka SSR aksesi-aksesi kelapasawit Kamerun. Bahan tanaman yang digunakan 49 aksesi kelapa sawitKamerun, Afrika yang ditanam di Kebun Sumber Daya Genetik (SDG)Sawit Sijunjung, Sumatera Barat. DNA genomik diisolasi dari tiapindividu aksesi menggunakan protokol isolasi DNA untuk tanamanbergetah. DNA dianalisis menggunakan 20 marka SSR. Dendrogramkekerabatan dikonstruksi menggunakan metode Unweighted Pair GroupMethod Arithmetic (UPGMA) melalui software NTSYS-pc (NumericalTaxonomy and Multivariate Analysis System) versi 2.1-pc. Hasil penelitianmenunjukkan nilai polimorfisme information content (PIC) marka SSRtinggi sebesar 0,80 (berkisar 0,63-0,91). Jumlah alel yang terdeteksi permarka SSR berkisar antara 4-15 alel per lokus SSR (rata-rata 8,75).Analisis filogenetik 49 aksesi menghasilkan diversitas genetik 12,5-54,72% (kemiripan genetik 55,28-87,50%). Pada diversitas genetik54,72%, aksesi Kamerun terbagi menjadi tujuh kelompok masing-masingterdiri dari 9, 28, 4, 2, 1, 2, dan 3 aksesi. Aksesi dengan diversitas genetiktinggi dan berada pada klaster berbeda, potensial digunakan sebagai calontetua dalam program pemuliaan kelapa sawit.Kata kunci: Elaeis guineensis Jacq., diversitas genetik, plasma nutfah,marka SSRABSTRACTGenetic diversity of the Indonesian oil palm collection is very low.To improve their genetic variability, exploration from the oil palm centerof origins has been done in Kamerun. The objectives of this study were todetermine genetic and polymorphism level of the SSR markers Cameroon-originated oil palm accessions. Genetic materials used were 49 Cameroon-originated oil palm accessions collected at Sijunjung Oil Palm GermplamCollection Station, West Sumatera. Genomic DNA was isolated using aprotocol for isolating DNA from leaves rich with latex. DNA was analyzedusing 20 SSR markers. A dendogram was constructed using theUnweighted Pair Group Method Arithmetic (UPGMA) method through theNumerical Taxonomy and Multivariate Analysis System software(NTSYS-pc) version 2.1-pc. Results showed that the polimorfismeinformation content (PIC) values of the SSR markers used was high, 0.80(range from 0.63-0.91). The average number of the SSR alleles detectedwas also high, 8.75 alleles (range from 4-15 alleles per SSR locus).Phylogenetic analysis of the 49 oil palm accessions resulted geneticdiversity of 12.5-54.72% (genetic similarity of 55.28-87.50%). At geneticdiversity 54.72%, the 49 accessions were divided into seven clusters, eachconsisted of 9, 28, 4, 2, 1, 2, and 3 accesions, respectively. Accessionswith high genetic diversity and located at different clusters may be usefulas parent candidates in the future oil palm breeding programs.Key words: Elaeis guineensis Jacq., genetic diversity, germplasm, SSRmarkers
Konstruksi Pustaka Genom Kakao (Theobroma cacao L.) untuk Sekuensing Genom Total Menggunakan Next Generation Sequencing HiSeq2000 Tasma, I Made; Satyawan, Dani; Rijzaani, Habib; Rubiyo, Rubiyo
Jurnal Tanaman Industri dan Penyegar Vol 3, No 2 (2012): Buletin Riset Tanaman Rempah Dan Aneka Tanaman Industri
Publisher : Pusat Penelitian dan Pengembangan Perkebunan

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Abstract

Pemuliaan kakao secara konvensional memerlukan waktu panjang (10-15 tahun). Pemanfaatan marka DNA akan memperpendek siklus pemuliaan kakao. Tujuan penelitian ini adalah mengkonstruksi pustaka genom tiga genotipe kakao yang dapat digunakan untuk sekuensing genom total kakao menggunakan NGS HiSeq2000 dan mendapatkan data resekuen genom total tiga genotipe kakao.  Bahan tanaman terdiri dari tiga klon unggul kakao (ICCR02, ICCR04, dan SUL02) diperoleh dari Balittri, Pakuwon.  DNA genomik diisolasi dari daun muda sebagai bahan konstruksi pustaka genom total. Sekuensing pustaka dilakukan pada mesin HiSeq2000 mengikuti protokol dari Illumina. Pustaka genom yang telah berhasil dikonstruksi berukuran 300 pasang basa (bp) masing-masing dengan konsentrasi 14,70 ng/µL (ICCR02), 15,20 ng/µL (ICCR04), dan 12,90 ng/µL (SUL02). Ukuran dan konsentrasi pustaka genom yang dihasilkan sangat ideal untuk sekuensing menggunakan HiSeq2000. Sekuensing ketiga genom menghasilkan data sekuen 52,9 x 109 bp.  Klaster DNA pustaka genom memiliki nilai Q scores>30 (75,0%) dengan tingkat kesalahan pembacaan basa rendah (1,47%).  Nilai densitas klaster, persen klaster PF, intensitas basa, persen phasing, dan persen prephasing menunjukkan kualitas klaster pustaka genom ketiga genotipe kakao termasuk kategori pustaka ideal. Data sekuen yang dihasilkan juga sangat ideal untuk identifikasi marka SNP genom kakao. Koleksi marka SNP digunakan untuk identifikasi gen pengendali karakter penting kakao dan pemuliaan berbasis marka DNA untuk memperpendek siklus pemuliaan kakao. Genomic Library Construction Of Cocoa (Theobroma Cacao L.) For Whole Genome Sequensing Using A Next Generation Sequencer Hiseq2000Conventional cocoa breeding is slow and takes about 10-15 years to complete a breeding cycle. Applying genomic technology using DNA markers will significantly decrease cocoa breeding cycle. The objectives of this study were to construct cocoa whole genome genomic libraries to be used for resequencing the whole genome of cocoa and obtain whole genome resequence data of three cocoa genotypes. Three Indonesian cocoa genotypes (ICCR02, ICRR04, and SUL02) were used. DNA genomic was isolated from young leaf and used to construct genomic DNA libraries and generate DNA clusters. DNA clusters were sequenced using a HiSeq2000 platform. The whole genome libraries of the cocoa genotypes were successfully constructed. The library size was 300 bp with concentrations of 14.70 ng/µL (ICCR02), 15.20 ng/µL (ICCR04), and 12.90 ng/µL (SUL02), respectively. The genomic library size and concentrations are suitable for sequencing study using the NGS HiSeq2000. Total sequencing output obtained was 52.9 x 109 bp. The genomic library clusters resulted during the sequencing process demonstrated the Q scores > 30 of 75.0% with low error sequencing rate of 1.47%. Cluster densities, percentage of cluster PF, base intensity, and percentage of phasing and prephasing indicated the cluster quality of the genomic libraries is classified as an ideal one to be used for resequencing study using NGS HiSeq2000. The resequence data were ideal for SNP marker discovery. SNP markers are used to identify economically important genes of cocoa and marker-aided cocoa breeding to decrase the cocoa breeding cycle.
PEMANFAATAN TEKNOLOGI SEKUENSING GENOM UNTUK MEMPERCEPAT PROGRAM PEMULIAAN TANAMAN Tasma, I Made
Jurnal Penelitian dan Pengembangan Pertanian Vol 34, No 4 (2015): Desember 2015
Publisher : Pusat Perpustakaan dan Penyebaran Teknologi Pertanian

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Sumber daya genetik (SDG) tanaman menyediakan materi dasar untuk program pemuliaan tanaman. Namun, baru sebagian kecil (<1%) koleksi SDG yang dimanfaatkan untuk pemuliaan tanaman. Karakterisasi SDG sudah banyak dilakukan dengan menggunakan karakter morfologi, namun metode ini lambat, menyita waktu, dan memerlukan banyak tenaga. Teknologi sekuensing modern menghasilkan peta genom rujukan suatu spesies tanaman yang   dapat mempercepat karakterisasi SDG menggunakan teknik next generation sequencing (NGS). Tulisan ini mengulas pemanfaatan teknologi sekuensing genom untuk karakterisasi, proteksi, dan pemanfaatan SDG untuk mempercepat program pemuliaan tanaman. Di Indonesia, teknologi NGS telah dimanfaatkan sejak 2010 untuk resekuensing genom komoditas unggulan nasional seperti kedelai, kakao, jagung, dan cabai merah. Jutaan SNP dan Indel telah diidentifikasi pada setiap komoditas sebagai sumber daya pemuliaan yang bernilai tinggi. Sebagian kecil SNP/Indel tersebut berada pada protein coding region yang potensial untuk penemuan gen-gen unggul. Selain SNP yang diidentifikasi pada semua genotipe, ditemukan SNP pada genotipe tertentu (SNP unik). Koleksi SNP dalam jumlah besar ini digunakan untuk mensintesis SNP chip untuk genotyping SDG secara cepat dan komprehensif. Didukung data fenotipe, SNP chip bermanfaat untuk melabel gen-gen unggul. Marka SNP yang berpautan dengan karakter unggul digunakan untuk menyeleksi individu pembawa karakter unggul tersebut. Dengan teknologi NGS, perakitan VUB tanaman dapat dilakukan lebih cepat, akurat, dan efisien. Dengan demikian, teknologi NGS dapat memfasilitasi karakterisasi dan pemanfaatan SDG untuk mem-percepat program pemuliaan tanaman.
Co-Authors Ahmad Warsun Budi Santosa Dani Satyawan HABIB RIJZAANI Muhamad Yunus Rubiyo Rubiyo SEKAR ARUMSARI, SEKAR