Cekaman kekeringan adalah salah satu faktor pembatas penting pada peningkatan produksi padi di In... more Cekaman kekeringan adalah salah satu faktor pembatas penting pada peningkatan produksi padi di Indonesia. Salah satunya cara pemecahannya adalah identifikasi gen-gen yang dapat digunakan untuk memperbaiki varietas padi toleran terhadap cekaman kekeringan. Sejumlah galur padi transgenic kultivar Nipponbare yang membawa vector penanda aktivasi telah dihasilkan dari penelitian sebelumnya. Untuk mengetahui apakah vector penanda aktivasi yang telah terintegrasi menghasilkan fenotipik peningkatan fungsi gen dalam responnya terhadap kekeringan, galur-galur tersebut perlu untuk dianalisis secara genotipk dan fenotipik. Tujuan penelitian ini adalah untuk mengetahui keragaman fenotipik akar galur-galur transgenik generasi T1 yang toleran terhadap cekaman kekeringan dan mendapatkan galur-galur yang stabil membawa penanda aktivasi. Bahan yang digunakan dalam penelitian adalah 47 individu galur transgenik, dua galur cek toleran (IRAT112 dan Cabacu), galur cek peka (IR64) dan tipe liar (Nipponba...
Overexpression of alanine aminotransferase (AlaAT) gene can improve nitrogen use efficiency (NUE)... more Overexpression of alanine aminotransferase (AlaAT) gene can improve nitrogen use efficiency (NUE) in plants. The previous isolated AlaAT genes cannot be freely applied to generate NUE plants due to IPR restriction. Therefore, isolation of the gene from targeted monocot and dicot plants is necessary. The objectives of this study were to isolate AlaAT genes from barley, foxtail millet, cucumber, and tomato and analyze their homology to other isolated AlaAT genes in sequence databases (gene bank). Total RNA was isolated from roots of barley, foxtail millet, cucumber, and tomato, and then converted into cDNA using reverse transcription method. The cDNA was then cloned into pGEM®-T Easy plasmid and the verified clones were sequenced. The amino acid sequences were analyzed for their homologies using Clustal Omega software and phylogenetic tree was constructed. The results showed that the amino acids of AlaAT gene from barley was different from AlaAT genes of tomato and cucumber with homol...
Drought stress is a major constrain which could influence rice productivity. Enhanced Response to... more Drought stress is a major constrain which could influence rice productivity. Enhanced Response to ABA1 (ERA1) gene encoding a β-subunit farnesyltransferase enzyme plays a role to control sensitivity of the guard cells to abscisic acid (ABA), hence regulating drought stress response in plant species including rice. This study aimed to clone the OsERA1 gene into expression vector, introduce it into rice plant, and confirm the positive OsERA1-rice plants conferring drought tolerance. This study was initiated by isolation of the OsERA1 gene from rice cDNAs and cloned it to an expression vector cassette, pCAMBIA1301. The cassette harboring OsERA1 gene was introduced into rice plant cv. Nipponbare mediated by Agrobacterium tumefaciens strain LBA4404.Putative transgenic lines were detected using PCR and Southern blot analyses to confirm the inserted transgene and the positive lines were assayed their tolerance to drought. The OsERA1 gene was successfully isolated and constructed into expre...
Kabupaten Enrekang merupakan sentra penanaman padi lokal Aromatik, hanya saja padi lokal tersebu... more Kabupaten Enrekang merupakan sentra penanaman padi lokal Aromatik, hanya saja padi lokal tersebut memiliki umur yang panjang dan produktivitasnya masih rendah. Salah satu cara untuk memperbaiki karakter padi lokal adalah dengan menyilangkan dengan padi unggul nasional. Penelitian ini bertujuan untuk memperoleh bibit padi lokal aromatik yang memiliki karakter seperti padi unggul nasional yaitu berumur pendek, memiliki aoma, potensi produksi tinggi dan tinggi berkisar 100 cm. Penelitian ini dilakukan pada bulan Maret – September 2014. Persilangan dilakukan antara padi Aromatik Lokal Kab. Enrekang yaitu Pare Mandoti, Pare Pallan, Pare Pinjan dan Pare Lotong dengan varietas unggul Ciherang. Hasil penelitian hingga tahap ini telah berhasi diperoleh generasi Fi hasil persilangan dan akan dikarakterisasi secara morfologi dan molecular untuk mendapatkan bibit tanaman padi Aromatik Unggul. Dari hasil penelitian dapat disimpulkan bahwa: 1. Telah berhasil dilakukan persilangan antara padi l...
Inpari HDB variety is resistant to bacterial leaf blight (BLB) disease, but due to its tall statu... more Inpari HDB variety is resistant to bacterial leaf blight (BLB) disease, but due to its tall stature, the variety is susceptible to lodging. Inpari HDB with semidwarf stature, therefore, is of high interest for lodging resistant performance. sd1 gene encoding GA20ox-2 enzyme is one of the genes responsible for imparting semidwarf stature of rice. In previous study, sd1 mutant rice cv. Kitaake (K15) was developed by using CRISPR/Cas9 technology. The objective of this study was to analyze molecularly and phenotypically F2 lines containing sd1 mutant gene resulted from a cross between Inpari HDB and K15 to develop semidwarf Inpari HDB rice variety. Thirty F2 Inpari HDB/K15 lines were analyzed at molecular level using DNA sequencing method together with phenotypic assessment of the lines to verify the integration of sd1 mutant gene. DNA sequencing analysis showed that 9 out of the 30 F2 Inpari HDB/K15 lines were sd1 mutants. The remaining F2 lines contained 17 heterozygotes and 4 nonmuta...
Yellow leaf curl disease caused by Pepper yellow leaf curl virus (PepYLCV), member of geminivirus... more Yellow leaf curl disease caused by Pepper yellow leaf curl virus (PepYLCV), member of geminiviruses group, is responsible for heavy yield losses for chili pepper production. Resistant genes which can cause immunity to the disease have not been found in germplasm collection. The aim of the research was to edit proliferating cell nuclear antigen (PCNA) gene by using CRISPR/Cas9 technology for developing plant resistance against geminivirus in chili pepper. A CRISPR/Cas9 plasmid cassette construct harboring the guide RNA of PCNA gene was constructed by Golden Gate cloning strategy. The construct was then introduced into chili genome via in planta method using Agrobacterium tumefaciens EHA105. The transformed plants were bioassayed by virus inoculation and confirmed using PCR and DNA sequencing to identify a mutagenesis event in PCNA gene target. The results showed that CRISPR/Cas9 plasmid cassette harboring gRNA of PCNA gene was successfully constructed. In planta transformation using ...
Tomato leaf curl virus (ToLCV) merupakan salah satu virus dalamgenus Begomovirus, famili Geminivi... more Tomato leaf curl virus (ToLCV) merupakan salah satu virus dalamgenus Begomovirus, famili Geminiviridae, yang menyebabkanpenyakit keriting daun pada tomat. Informasi tentang keragamangenetik ToLCV bermanfaat dalam perakitan tanaman tahan. Kemajuandi bidang biologi molekuler telah menghasilkan beberapa teknikyang dapat digunakan untuk analisis genetik Begomovirus. Teknikmolekuler yang banyak diaplikasikan ialah polymerase chainreaction (PCR). Teknik ini sangat sensitif dan spesifik untukmendeteksi Begomovirus pada tingkat DNA. PCR juga dapatdigunakan untuk mengidentifikasi tingkat keragaman genetik virus.Teknik PCR telah digunakan untuk mendeteksi Begomovirus padatomat (ToLCV) dari sentra produksi di Jawa Timur, Jawa Tengah,Jawa Barat, DI Yogyakarta, dan Sumatera. Kombinasi teknik PCRdengan restriction fragment length polymorphism (RFLP) jugadapat digunakan untuk mengidentifikasi keragaman genetik Begomovirus.Selain itu, teknik sekuensing DNA dapat diaplikasikanuntuk mempelajari ident...
Aromatic rice varieties have some weaknesses such as low productivity, and less resistant to pest... more Aromatic rice varieties have some weaknesses such as low productivity, and less resistant to pests and diseases. This study aimed to obtain homozygous strain of BC5F2 Ciherang aromatic through the identification of aroma gene (mutated badh2) and properties of the aroma. Ciherang paddy (nonaromatic paddy) was used as the female parent, whereas Mentik Wangi paddy (aromatic paddy) was used as the male parent. The experiment was conducted in BC5F2 because it is expected to generate plants with properties 98.4% close to female parent. The DNA from five strains of paddy plants BC5F2Ciherang X Mentik Wangi was isolated by a modified CTAB method. The concentration of DNA was determined by measuring absorbance at 260 nm wavelength, while its purity was determined from the ratio of the absorbance at a wavelength of 260/280 nm. PCR-based molecular selection was done by using the Bradbury primers. PCR results showed that of the 250 samples, there were 66 samples had DNA fragment of the same siz...
The development of submergence tolerant rice varieties is urgently required to maintain the stabi... more The development of submergence tolerant rice varieties is urgently required to maintain the stability of future food production, to anticipate the unpredictable global climate changes. Due to in-economical agronomic traits of native submergence tolerant varieties for large scale cultivation, submergence tolerance gene (sub1) must be introduced into popular high-yielding rice variety, such as Ciherang. To develop new submergence tolerant variety with good agronomic traits as those of Ciherang, in this research, submergence tolerance gene (sub1) was introduced into Ciherang variety. To avoid strict GMO regulation, gene introduction was carried out through site-directed crossing. Donor sub1 was crossed with Ciherang host. The selected F1 progenies were further backcrossed to Ciherang 4 x to obtain BC5F1 progeny having ~98% agronomic traits of those of Ciherang. In every cross/backcross generation, submergence test was performed, followed by sub1 marker-assisted PCR. F1 and BC1F1 submer...
ABSTRAKKemiri sunan merupakan salah satu tanaman penghasil biodieseldengan potensi yang sangat be... more ABSTRAKKemiri sunan merupakan salah satu tanaman penghasil biodieseldengan potensi yang sangat besar disamping pemanfaatannya sebagaitanaman konservasi. Minyak kemiri sunan mengandung racun sehinggatidak dapat dikonsumsi. Dikatakan bahwa asam α-eleostearat dengankandungan 50% dalam minyak merupakan senyawa yang mengakibatkanminyak kemiri sunan beracun. Sebagai tanaman yang potensial, makasangat diperlukan informasi lengkap tentang tanaman tersebut, termasukanalisis DNA. Berbagai teknik dapat dilakukan untuk mengisolasi DNAtergantung dari jenis tanaman, organ tanaman, atau jaringan tanaman yangdigunakan. Penelitian ini bertujuan untuk mendapatkan teknik isolasi danpurifikasi DNA yang efektif dan efisien, sehingga bisa mengurangi biayadan penghematan waktu dalam pengerjaan di laboratorium. Penelitiandilaksanakan di Laboratorium Biologi Molekuler, Balai Besar Penelitiandan Pengembangan Bioteknologi dan Sumberdaya Genetik Pertanian (BBBiogen), Bogor pada bulan Juli-September 2010. Mater...
Pemanfaatan tanaman kentang produk rekayasa genetik (PRG) dalam pemuliaan tanaman melalui persila... more Pemanfaatan tanaman kentang produk rekayasa genetik (PRG) dalam pemuliaan tanaman melalui persilangan dengan Atlantic dan Granola telah menghasilkan enam galur PRG hasil silangan yang terseleksi. Sebelum komersialisasi, kentang PRG harus dikaji keamanan pangan dan lingkungannya. Penulisan bertujuan memberikan informasi mengenai tanaman kentang PRG di Indonesia yang tahan terhadap penyakit busuk daun Phytophthora infestans dan telah dinyatakan aman untuk di-konsumsi oleh manusia. Analisis stabilitas menunjukkan bahwa gen RB stabil terintegrasi selama empat generasi klonal ber-urutan dalam genom tanaman kentang PRG dengan satu sisipan gen. Hasil studi komposisi dan nutrisi, glikoalkaloid total, dan anti nutrisi pada kentang PRG Katahdin SP951 dan galur-galur silangannya bersifat sepadan dengan Katahdin non-PRG. Studi toksisitas menunjukkan bahwa pemberian pakan suspensi umbi kentang dan suspensi tepung kentang Katahdin SP951 dan galur-galur silangan tidak berdampak terhadap mortalitas...
Jurnal Agronomi Indonesia (Indonesian Journal of Agronomy), 2018
The genetic transformation via pollen-tube pathway in jatropha is the first alternative method th... more The genetic transformation via pollen-tube pathway in jatropha is the first alternative method that was applied in this plant. The objective of the research was to study fruit set and germination of three genotypes of jatropha following direct transformations via pollen-tube pathways. The research was conducted during April 2014 until January 2015 at jatropa’s experimental field, Pasuruan, and at green house of ICABIOGRAD, Bogor. Three genotypes of jatropha i.e., IP3A, IP3P and JcUMM18 were used. In the first experiment, split plot design was used where 3 levels of DNA plasmid concentration as a main plot and 5 levels of stigma-drip time of DNA plasmid as a subplot. In the second experiment randomized block design was used with single factor consisting of 15 combinations of concentration and stigma-drip time of DNA plasmid as treatments and control. The results demonstrated that interaction between concentration and application time of DNA plasmid did not significantly affect fruit ...
Rice (Oryza sativa L.) is important staple crop in Indonesia. Food demand that continues to rise ... more Rice (Oryza sativa L.) is important staple crop in Indonesia. Food demand that continues to rise while inadequate land could be managed by assembling superior cultivar using CRISPR-Cas9 system method. Editing the genome by mutating the GA20ox-2 gene could improve both crop yield and ability to thrive in marginal land (drought). This experiment aims to obtain non-transgenic mutant plants (non Cas9 and hpt genes), gain information on GA20ox-2 gene expression levels, and study the tolerance levels of the CRISPR /Cas9-OsGA20ox-2 mutant lines 'Kitaake' T2 generation against drought conditions. Planting material using a mutant gene GA20ox 2 ‘Kitaake’ (K23.1, K15, K29.1, K19.1) and wild-type comparison. From 20 plants, respectively the K23.1, K15, K29.1, and K19.1 lines have 50%, 50%, 0%, and 45% of non-transgenic plants. DNA mutations in the form of deletion 44 bases (K23.1, K29.1, K19.1) and insertion of two bases (K15) are transcribed into RNA. The transcription results in a num...
ABSTRACTBegomoviruses infection has been reported and found in several important vegetable crops ... more ABSTRACTBegomoviruses infection has been reported and found in several important vegetable crops included tomato. Nevertheless, the information of detection and identification of Begomovirus infecting tomato plants in some of tomato production areas using PCR technique has not been widely reported. The objective of this research was to detect Begomovirus infecting tomatoes in some of tomato production areas of East Java, Central Java, Special Province of Jogjakarta and West Java using PCR technique with degenerate and specific primers. PCR amplification of Begomovirus genome was conducted by using a pair of degenerate primers, i.e. PAL1v1978-F and PAR1c715-R and specific primer, AV1-F/R. For confirmation the virus, it was conducted a virus transmission from the symptomed plant tomato sampel to healthy plants by using whiteflies vector. The results of this research showed that the symptomed plants collected from severa...
Transformasi gen Glu-1Dx5, pengendali utama karakter elastisitas dan daya mengembang adonan dari ... more Transformasi gen Glu-1Dx5, pengendali utama karakter elastisitas dan daya mengembang adonan dari gandum, telah berhasil ditransfer ke dalam genom tanaman padi kultivar Fatmawati dengan menggunakan penembakan partikel, dengan tujuan untuk memperbaiki kualitas adonan tepung beras. Galur-galur harapan telah diperoleh, tetapi karena telah mengalami penyerbukan sendiri selama 1-2 generasi yang menyebabkan transgen mengalami segregasi, maka diperlukan upaya pendeteksian transgen pada populasi putative transgenik ini. Upaya ini dapat dilakukan, antara lain dengan menggunakan teknik Polymerase Chain Reaction (PCR) yang memungkinkan perbanyakan fragmen DNA yang spesifik (gen) secara cepat dalam jumlah banyak. Percobaan ini bertujuan untuk mendapatkan tanaman padi transgenik yang memiliki gen Glu-1Dx5 pada dua generasi yang sedang bersegregasi. DNA genom dari 149 tanaman padi (generasi T1 sebanyak 14 tanaman, generasi T2 sebanyak 134 tanaman, dan satu tanaman non-transgenik) telah diekstraks...
Methods in molecular biology (Clifton, N.J.), 2018
Genetic transformation of maize inbred genotypes remains non-routine for many laboratories due to... more Genetic transformation of maize inbred genotypes remains non-routine for many laboratories due to variations in cell competency to induce embryogenic callus, as well as the cell's ability to receive and incorporate transgenes into the genome. This chapter describes two transformation protocols using Agrobacterium- and biolistic-mediated methods for gene delivery. Immature zygotic embryos of maize inbred B104, excised from ears harvested 10-14 days post pollination, are used as starting explant material. Disarmed Agrobacterium strains harboring standard binary vectors and the biolistic gun system Bio-Rad PDS-1000/He are used as gene delivery systems. The herbicide resistant bar gene and selection agent bialaphos are used for identifying putative transgenic type I callus events. Using the step-by-step protocols described here, average transformation frequencies (number of bialaphos resistant T0 callus events per 100 explants infected or bombarded) of 4% and 8% can be achieved usin...
Cekaman kekeringan adalah salah satu faktor pembatas penting pada peningkatan produksi padi di In... more Cekaman kekeringan adalah salah satu faktor pembatas penting pada peningkatan produksi padi di Indonesia. Salah satunya cara pemecahannya adalah identifikasi gen-gen yang dapat digunakan untuk memperbaiki varietas padi toleran terhadap cekaman kekeringan. Sejumlah galur padi transgenic kultivar Nipponbare yang membawa vector penanda aktivasi telah dihasilkan dari penelitian sebelumnya. Untuk mengetahui apakah vector penanda aktivasi yang telah terintegrasi menghasilkan fenotipik peningkatan fungsi gen dalam responnya terhadap kekeringan, galur-galur tersebut perlu untuk dianalisis secara genotipk dan fenotipik. Tujuan penelitian ini adalah untuk mengetahui keragaman fenotipik akar galur-galur transgenik generasi T1 yang toleran terhadap cekaman kekeringan dan mendapatkan galur-galur yang stabil membawa penanda aktivasi. Bahan yang digunakan dalam penelitian adalah 47 individu galur transgenik, dua galur cek toleran (IRAT112 dan Cabacu), galur cek peka (IR64) dan tipe liar (Nipponba...
Overexpression of alanine aminotransferase (AlaAT) gene can improve nitrogen use efficiency (NUE)... more Overexpression of alanine aminotransferase (AlaAT) gene can improve nitrogen use efficiency (NUE) in plants. The previous isolated AlaAT genes cannot be freely applied to generate NUE plants due to IPR restriction. Therefore, isolation of the gene from targeted monocot and dicot plants is necessary. The objectives of this study were to isolate AlaAT genes from barley, foxtail millet, cucumber, and tomato and analyze their homology to other isolated AlaAT genes in sequence databases (gene bank). Total RNA was isolated from roots of barley, foxtail millet, cucumber, and tomato, and then converted into cDNA using reverse transcription method. The cDNA was then cloned into pGEM®-T Easy plasmid and the verified clones were sequenced. The amino acid sequences were analyzed for their homologies using Clustal Omega software and phylogenetic tree was constructed. The results showed that the amino acids of AlaAT gene from barley was different from AlaAT genes of tomato and cucumber with homol...
Drought stress is a major constrain which could influence rice productivity. Enhanced Response to... more Drought stress is a major constrain which could influence rice productivity. Enhanced Response to ABA1 (ERA1) gene encoding a β-subunit farnesyltransferase enzyme plays a role to control sensitivity of the guard cells to abscisic acid (ABA), hence regulating drought stress response in plant species including rice. This study aimed to clone the OsERA1 gene into expression vector, introduce it into rice plant, and confirm the positive OsERA1-rice plants conferring drought tolerance. This study was initiated by isolation of the OsERA1 gene from rice cDNAs and cloned it to an expression vector cassette, pCAMBIA1301. The cassette harboring OsERA1 gene was introduced into rice plant cv. Nipponbare mediated by Agrobacterium tumefaciens strain LBA4404.Putative transgenic lines were detected using PCR and Southern blot analyses to confirm the inserted transgene and the positive lines were assayed their tolerance to drought. The OsERA1 gene was successfully isolated and constructed into expre...
Kabupaten Enrekang merupakan sentra penanaman padi lokal Aromatik, hanya saja padi lokal tersebu... more Kabupaten Enrekang merupakan sentra penanaman padi lokal Aromatik, hanya saja padi lokal tersebut memiliki umur yang panjang dan produktivitasnya masih rendah. Salah satu cara untuk memperbaiki karakter padi lokal adalah dengan menyilangkan dengan padi unggul nasional. Penelitian ini bertujuan untuk memperoleh bibit padi lokal aromatik yang memiliki karakter seperti padi unggul nasional yaitu berumur pendek, memiliki aoma, potensi produksi tinggi dan tinggi berkisar 100 cm. Penelitian ini dilakukan pada bulan Maret – September 2014. Persilangan dilakukan antara padi Aromatik Lokal Kab. Enrekang yaitu Pare Mandoti, Pare Pallan, Pare Pinjan dan Pare Lotong dengan varietas unggul Ciherang. Hasil penelitian hingga tahap ini telah berhasi diperoleh generasi Fi hasil persilangan dan akan dikarakterisasi secara morfologi dan molecular untuk mendapatkan bibit tanaman padi Aromatik Unggul. Dari hasil penelitian dapat disimpulkan bahwa: 1. Telah berhasil dilakukan persilangan antara padi l...
Inpari HDB variety is resistant to bacterial leaf blight (BLB) disease, but due to its tall statu... more Inpari HDB variety is resistant to bacterial leaf blight (BLB) disease, but due to its tall stature, the variety is susceptible to lodging. Inpari HDB with semidwarf stature, therefore, is of high interest for lodging resistant performance. sd1 gene encoding GA20ox-2 enzyme is one of the genes responsible for imparting semidwarf stature of rice. In previous study, sd1 mutant rice cv. Kitaake (K15) was developed by using CRISPR/Cas9 technology. The objective of this study was to analyze molecularly and phenotypically F2 lines containing sd1 mutant gene resulted from a cross between Inpari HDB and K15 to develop semidwarf Inpari HDB rice variety. Thirty F2 Inpari HDB/K15 lines were analyzed at molecular level using DNA sequencing method together with phenotypic assessment of the lines to verify the integration of sd1 mutant gene. DNA sequencing analysis showed that 9 out of the 30 F2 Inpari HDB/K15 lines were sd1 mutants. The remaining F2 lines contained 17 heterozygotes and 4 nonmuta...
Yellow leaf curl disease caused by Pepper yellow leaf curl virus (PepYLCV), member of geminivirus... more Yellow leaf curl disease caused by Pepper yellow leaf curl virus (PepYLCV), member of geminiviruses group, is responsible for heavy yield losses for chili pepper production. Resistant genes which can cause immunity to the disease have not been found in germplasm collection. The aim of the research was to edit proliferating cell nuclear antigen (PCNA) gene by using CRISPR/Cas9 technology for developing plant resistance against geminivirus in chili pepper. A CRISPR/Cas9 plasmid cassette construct harboring the guide RNA of PCNA gene was constructed by Golden Gate cloning strategy. The construct was then introduced into chili genome via in planta method using Agrobacterium tumefaciens EHA105. The transformed plants were bioassayed by virus inoculation and confirmed using PCR and DNA sequencing to identify a mutagenesis event in PCNA gene target. The results showed that CRISPR/Cas9 plasmid cassette harboring gRNA of PCNA gene was successfully constructed. In planta transformation using ...
Tomato leaf curl virus (ToLCV) merupakan salah satu virus dalamgenus Begomovirus, famili Geminivi... more Tomato leaf curl virus (ToLCV) merupakan salah satu virus dalamgenus Begomovirus, famili Geminiviridae, yang menyebabkanpenyakit keriting daun pada tomat. Informasi tentang keragamangenetik ToLCV bermanfaat dalam perakitan tanaman tahan. Kemajuandi bidang biologi molekuler telah menghasilkan beberapa teknikyang dapat digunakan untuk analisis genetik Begomovirus. Teknikmolekuler yang banyak diaplikasikan ialah polymerase chainreaction (PCR). Teknik ini sangat sensitif dan spesifik untukmendeteksi Begomovirus pada tingkat DNA. PCR juga dapatdigunakan untuk mengidentifikasi tingkat keragaman genetik virus.Teknik PCR telah digunakan untuk mendeteksi Begomovirus padatomat (ToLCV) dari sentra produksi di Jawa Timur, Jawa Tengah,Jawa Barat, DI Yogyakarta, dan Sumatera. Kombinasi teknik PCRdengan restriction fragment length polymorphism (RFLP) jugadapat digunakan untuk mengidentifikasi keragaman genetik Begomovirus.Selain itu, teknik sekuensing DNA dapat diaplikasikanuntuk mempelajari ident...
Aromatic rice varieties have some weaknesses such as low productivity, and less resistant to pest... more Aromatic rice varieties have some weaknesses such as low productivity, and less resistant to pests and diseases. This study aimed to obtain homozygous strain of BC5F2 Ciherang aromatic through the identification of aroma gene (mutated badh2) and properties of the aroma. Ciherang paddy (nonaromatic paddy) was used as the female parent, whereas Mentik Wangi paddy (aromatic paddy) was used as the male parent. The experiment was conducted in BC5F2 because it is expected to generate plants with properties 98.4% close to female parent. The DNA from five strains of paddy plants BC5F2Ciherang X Mentik Wangi was isolated by a modified CTAB method. The concentration of DNA was determined by measuring absorbance at 260 nm wavelength, while its purity was determined from the ratio of the absorbance at a wavelength of 260/280 nm. PCR-based molecular selection was done by using the Bradbury primers. PCR results showed that of the 250 samples, there were 66 samples had DNA fragment of the same siz...
The development of submergence tolerant rice varieties is urgently required to maintain the stabi... more The development of submergence tolerant rice varieties is urgently required to maintain the stability of future food production, to anticipate the unpredictable global climate changes. Due to in-economical agronomic traits of native submergence tolerant varieties for large scale cultivation, submergence tolerance gene (sub1) must be introduced into popular high-yielding rice variety, such as Ciherang. To develop new submergence tolerant variety with good agronomic traits as those of Ciherang, in this research, submergence tolerance gene (sub1) was introduced into Ciherang variety. To avoid strict GMO regulation, gene introduction was carried out through site-directed crossing. Donor sub1 was crossed with Ciherang host. The selected F1 progenies were further backcrossed to Ciherang 4 x to obtain BC5F1 progeny having ~98% agronomic traits of those of Ciherang. In every cross/backcross generation, submergence test was performed, followed by sub1 marker-assisted PCR. F1 and BC1F1 submer...
ABSTRAKKemiri sunan merupakan salah satu tanaman penghasil biodieseldengan potensi yang sangat be... more ABSTRAKKemiri sunan merupakan salah satu tanaman penghasil biodieseldengan potensi yang sangat besar disamping pemanfaatannya sebagaitanaman konservasi. Minyak kemiri sunan mengandung racun sehinggatidak dapat dikonsumsi. Dikatakan bahwa asam α-eleostearat dengankandungan 50% dalam minyak merupakan senyawa yang mengakibatkanminyak kemiri sunan beracun. Sebagai tanaman yang potensial, makasangat diperlukan informasi lengkap tentang tanaman tersebut, termasukanalisis DNA. Berbagai teknik dapat dilakukan untuk mengisolasi DNAtergantung dari jenis tanaman, organ tanaman, atau jaringan tanaman yangdigunakan. Penelitian ini bertujuan untuk mendapatkan teknik isolasi danpurifikasi DNA yang efektif dan efisien, sehingga bisa mengurangi biayadan penghematan waktu dalam pengerjaan di laboratorium. Penelitiandilaksanakan di Laboratorium Biologi Molekuler, Balai Besar Penelitiandan Pengembangan Bioteknologi dan Sumberdaya Genetik Pertanian (BBBiogen), Bogor pada bulan Juli-September 2010. Mater...
Pemanfaatan tanaman kentang produk rekayasa genetik (PRG) dalam pemuliaan tanaman melalui persila... more Pemanfaatan tanaman kentang produk rekayasa genetik (PRG) dalam pemuliaan tanaman melalui persilangan dengan Atlantic dan Granola telah menghasilkan enam galur PRG hasil silangan yang terseleksi. Sebelum komersialisasi, kentang PRG harus dikaji keamanan pangan dan lingkungannya. Penulisan bertujuan memberikan informasi mengenai tanaman kentang PRG di Indonesia yang tahan terhadap penyakit busuk daun Phytophthora infestans dan telah dinyatakan aman untuk di-konsumsi oleh manusia. Analisis stabilitas menunjukkan bahwa gen RB stabil terintegrasi selama empat generasi klonal ber-urutan dalam genom tanaman kentang PRG dengan satu sisipan gen. Hasil studi komposisi dan nutrisi, glikoalkaloid total, dan anti nutrisi pada kentang PRG Katahdin SP951 dan galur-galur silangannya bersifat sepadan dengan Katahdin non-PRG. Studi toksisitas menunjukkan bahwa pemberian pakan suspensi umbi kentang dan suspensi tepung kentang Katahdin SP951 dan galur-galur silangan tidak berdampak terhadap mortalitas...
Jurnal Agronomi Indonesia (Indonesian Journal of Agronomy), 2018
The genetic transformation via pollen-tube pathway in jatropha is the first alternative method th... more The genetic transformation via pollen-tube pathway in jatropha is the first alternative method that was applied in this plant. The objective of the research was to study fruit set and germination of three genotypes of jatropha following direct transformations via pollen-tube pathways. The research was conducted during April 2014 until January 2015 at jatropa’s experimental field, Pasuruan, and at green house of ICABIOGRAD, Bogor. Three genotypes of jatropha i.e., IP3A, IP3P and JcUMM18 were used. In the first experiment, split plot design was used where 3 levels of DNA plasmid concentration as a main plot and 5 levels of stigma-drip time of DNA plasmid as a subplot. In the second experiment randomized block design was used with single factor consisting of 15 combinations of concentration and stigma-drip time of DNA plasmid as treatments and control. The results demonstrated that interaction between concentration and application time of DNA plasmid did not significantly affect fruit ...
Rice (Oryza sativa L.) is important staple crop in Indonesia. Food demand that continues to rise ... more Rice (Oryza sativa L.) is important staple crop in Indonesia. Food demand that continues to rise while inadequate land could be managed by assembling superior cultivar using CRISPR-Cas9 system method. Editing the genome by mutating the GA20ox-2 gene could improve both crop yield and ability to thrive in marginal land (drought). This experiment aims to obtain non-transgenic mutant plants (non Cas9 and hpt genes), gain information on GA20ox-2 gene expression levels, and study the tolerance levels of the CRISPR /Cas9-OsGA20ox-2 mutant lines 'Kitaake' T2 generation against drought conditions. Planting material using a mutant gene GA20ox 2 ‘Kitaake’ (K23.1, K15, K29.1, K19.1) and wild-type comparison. From 20 plants, respectively the K23.1, K15, K29.1, and K19.1 lines have 50%, 50%, 0%, and 45% of non-transgenic plants. DNA mutations in the form of deletion 44 bases (K23.1, K29.1, K19.1) and insertion of two bases (K15) are transcribed into RNA. The transcription results in a num...
ABSTRACTBegomoviruses infection has been reported and found in several important vegetable crops ... more ABSTRACTBegomoviruses infection has been reported and found in several important vegetable crops included tomato. Nevertheless, the information of detection and identification of Begomovirus infecting tomato plants in some of tomato production areas using PCR technique has not been widely reported. The objective of this research was to detect Begomovirus infecting tomatoes in some of tomato production areas of East Java, Central Java, Special Province of Jogjakarta and West Java using PCR technique with degenerate and specific primers. PCR amplification of Begomovirus genome was conducted by using a pair of degenerate primers, i.e. PAL1v1978-F and PAR1c715-R and specific primer, AV1-F/R. For confirmation the virus, it was conducted a virus transmission from the symptomed plant tomato sampel to healthy plants by using whiteflies vector. The results of this research showed that the symptomed plants collected from severa...
Transformasi gen Glu-1Dx5, pengendali utama karakter elastisitas dan daya mengembang adonan dari ... more Transformasi gen Glu-1Dx5, pengendali utama karakter elastisitas dan daya mengembang adonan dari gandum, telah berhasil ditransfer ke dalam genom tanaman padi kultivar Fatmawati dengan menggunakan penembakan partikel, dengan tujuan untuk memperbaiki kualitas adonan tepung beras. Galur-galur harapan telah diperoleh, tetapi karena telah mengalami penyerbukan sendiri selama 1-2 generasi yang menyebabkan transgen mengalami segregasi, maka diperlukan upaya pendeteksian transgen pada populasi putative transgenik ini. Upaya ini dapat dilakukan, antara lain dengan menggunakan teknik Polymerase Chain Reaction (PCR) yang memungkinkan perbanyakan fragmen DNA yang spesifik (gen) secara cepat dalam jumlah banyak. Percobaan ini bertujuan untuk mendapatkan tanaman padi transgenik yang memiliki gen Glu-1Dx5 pada dua generasi yang sedang bersegregasi. DNA genom dari 149 tanaman padi (generasi T1 sebanyak 14 tanaman, generasi T2 sebanyak 134 tanaman, dan satu tanaman non-transgenik) telah diekstraks...
Methods in molecular biology (Clifton, N.J.), 2018
Genetic transformation of maize inbred genotypes remains non-routine for many laboratories due to... more Genetic transformation of maize inbred genotypes remains non-routine for many laboratories due to variations in cell competency to induce embryogenic callus, as well as the cell's ability to receive and incorporate transgenes into the genome. This chapter describes two transformation protocols using Agrobacterium- and biolistic-mediated methods for gene delivery. Immature zygotic embryos of maize inbred B104, excised from ears harvested 10-14 days post pollination, are used as starting explant material. Disarmed Agrobacterium strains harboring standard binary vectors and the biolistic gun system Bio-Rad PDS-1000/He are used as gene delivery systems. The herbicide resistant bar gene and selection agent bialaphos are used for identifying putative transgenic type I callus events. Using the step-by-step protocols described here, average transformation frequencies (number of bialaphos resistant T0 callus events per 100 explants infected or bombarded) of 4% and 8% can be achieved usin...
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Papers by Tri Joko Santoso