Articles

Pengaruh Perlakuan Ethyl Methane Sulfonate pada Tanaman Cabai (Capsicum annuum L.) dan Ketahanannya Terhadap Chilli Veinal Mottle Virus (ChiVMV) Manzila, Ifa; Hidayat, Sri Hendrastuti; Mariska, Ika; Sujiprihati, Sriani
Jurnal Agronomi Indonesia (Indonesian Journal of Agronomy) Vol 38, No 3 (2010): Jurnal Agronomi Indonesia
Publisher : Bogor Agricultural University

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Ethyl Methane Sulfonate (EMS) may induce mutation leading to somaclonal variation if it is used at the appropriate combination of EMS concentration and exposure time. Variation in somaclonal might be valuable as a source of resistance to plant pathogens including  plant viruses. This study was aimed 1) to determine the optimum EMS concentration and incubation time that may induce somaclonal variation in chilli pepper; and 2) to evaluate the resistance of the somaclone to ChiVMV  infection. Shoot-tip explants of five chilli pepper genotypes (Jatilaba, ICPN 12 no. 4, PBC495, Helem, and Gelora) were treated with EMS at combination of different concentrations (0.25%, 0.5%  1.0% and control),  and incubation time (15, 30, 60 min). Subsequently, each explant was grown  in multiplication media (MS media + 5 mg L-1 BAP + 0.5 mg L-1 TDZ), rooting media (MS  media +  1 mg L-1 NAA), and acclimatization media (mixture of soil : sand : compost  2:1:1 w/w). Our results showed that the higher EMS concentration and the longer incubation period the smaller the number of survive explants. The highest survival rate  20.4 %  was achieved with 0.5% EMS in combination with 60 min  incubation period. This treatment combination also showed induction of phenotypic variation. Two somaclonal plants derived from Gelora genotype, designated as somaclones K1 and K2,  survived until fruit development and maturation. A total of 245 progenies of K1 and 243 progenies of K2, respectively were evaluated for their resistance to ChiVMV infection through mechanical inoculation using ChiVMV-Cikabayan isolate. Following the detection of ChiVMV using DAS-ELISA, it was confirmed that  4.09% of the somaclonal progenies were  resistance to ChiVMV.   Keywords:  Capsicum annuum, ChiVMV, ethyl methane sulfonate, induce mutation, resistance
Characterization of B-glukosidase Enzyme from Vanilla Bean Setyaningsih, Dwi; Soehartono, Maggy T; Apriyantono, Anton; Mariska, Ika
Jurnal Teknologi Dan Industri Pangan Vol 18, No 2 (2007): Jurnal Teknologi dan Industri Pangan
Publisher : Departemen Ilmu dan Teknologi Pangan, IPB Indonesia bekerjasama dengan PATPI

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The Indonesian natural vanilla is know for having a unigue woody, smooky, and phenolic flavor. Development of the aroma and flavor vanilla was formed by the action of a hydrolytic enzyme B-glucosidase on glucovanillin. The objective of this research was to characterize vanilla B-glucosidase. The vanilla B-glucosidase activity was increased by detergent. The enzyme was found as heat labile. Scalding should be conducted at 400C for 2-3 minutes. The result from B-glucosidase activity in each part of vanilla and microscopic analisis of vanilla bean slice showed that the highest B-glucosidase activity and vanillin concentrations were found in the seed funicles and placental tissue the of vanilla bean. The activity of vanilla B-glucosidase was optimum at pH 6,0, and temperature of 400C, found as and activation energy was 5,78 kcal/mole. After 44 minutes incubation time at 400C. The activity was reduced down to 10%. The apparent of moleculer weight was 100-400 kDa according to gel setration (Sephacryl S-300) analysis. Key words : Vanilla planifolia, B-glucosidase
Maceration Process Optimation of Vanili (Vanilla Planifolia Andrews) from Modified Curing Setyaningsih, Dwi; Rusli, Meika S; ., Melawati; Mariska, Ika
Jurnal Teknologi Dan Industri Pangan Vol 17, No 2 (2006): Jurnal Teknologi dan Industri Pangan
Publisher : Departemen Ilmu dan Teknologi Pangan, IPB Indonesia bekerjasama dengan PATPI

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Abstract

Modified cured vanilla was processed to vanilla extract by maceration method. The aim of this research were to optimize the method of maceration, type of vanilla bean with highest vanillin content, extraction solvent composition,and other variables that could optimize the vanillin content and characterize the extract from half dried cured vanilla. The optimation used response surface method with 22 factorial and 23 factorial. One step of maceration could extract vanillin (average 2.3 g/l) much more than two steps maceration (average 2.1 g/l). Vanillin content of the half dried cured vanilla (average 0.98 g/l) was higher than cured vanilla 1 and cured vanilla 2 (average 0.41 g/l and 0.32 g/l). The suitable ethanol-water composition for half dried cured vanilla was 7:3 (vanillin content 1.78 g/l). The first optimation was conducted with two variables maceration time and sucrose concentrations. The maximum vanillin content of the first optimation was 4.5 g/l at maceration time of15.9 days and sucrose concentration of 7.3 g. The second optimation used two variables: maceration time and glycerol concentrations. The maximum vanillin content of the second optimation was 3.8 g/l at maceration time of 22 days and glycerol concentration 19.9 ml. The third optimation process used three variables:maceration time, sucrose concentrations and glycerol concentrations. The maximum vanillin content of the third optimation was 3.4 g/l at maceration time of 12 days sucrose concentration of 7 g, and glycerol concentration 4.7 ml. The characteristic of vanilla extract resulted from half dried cured vanilla maceration were vanillin content (3.4-4.5 g/l), total acid (380-410 ml 0.1 N NaOH/l), total ash (1.3-3.4 g/l), total soluble ash (0.8-2.9 g/l), alkalinity of total ash (462.6-536.7), alkalinity of soluble ash (139.1-216.5), and lead number (4.5-4.6). Key words : Vanilla planifolia, Optimization, vanilla ekstrak  
Peningkatan Toleransi Kedelai Sindoro terhadap Kekeringan Melalui Seleksi In Vitro Husni, Ali; Kosmiatin, M.; Mariska, Ika
Jurnal Agronomi Indonesia (Indonesian Journal of Agronomy) Vol 34, No 1 (2006): Jurnal Agronomi Indonesia
Publisher : Bogor Agricultural University

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In vitro selection of embryogenic cell mass is one alternative to improve drought tolerance in plants. Embryogenic cell callus of soybean were radiated by Gamma ray (400 rad) to produce mutation. The radiated cell  were tested with PEG (0, 10, 20 and 30 %) for drought stress tolerance. After selection, cells which tolerant to PEG were regenerated to produce somatic embryo structure, somatic seed and plantlet. Acclimatization was done in a greenhouse and analysis of proline was done at generation 1 (G1). The purpose of the experiment was to get soybean somatic seed which tolerant to drought stress. Results of experiment showed that 39.7 % embriogenic callus were produced. The higher the concentration of PEG, the higher the death of cell/callus. The rate of producing somatic embryo structure was 4.9 at 0 % PEG; 2.85 at 10 % PEG; 1.6 at 20% PEG and 0.6 at 30% PEG. Number of somatic seed which developed in regeneration medium (S11) were 79 from 0% PEG;  35 from 10% PEG; 29 from 20% PEG, and 15 from 30% PEG. Somatic seed produced 15 planlets from PEG 0%; 6 planlets from PEG10%; 4 planlets from PEG 20%. Based of proline content, all of G1 somaclones were more tolerant  than the mother plant.     Key words : Soybean, in vitro selection, PEG, regeneration, acclimatization and dry land.                     
Identifikasi Somaklon Padi Gajahmungkur, Towuti dan IR 64 Tahan Kekeringan Menggunakan Polyethylene Glycol Lestari, Endang Gati; Mariska, Ika
Jurnal Agronomi Indonesia (Indonesian Journal of Agronomy) Vol 34, No 2 (2006): Jurnal Agronomi Indonesia
Publisher : Bogor Agricultural University

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Abstract

The drought stress tolerant and high yielding rice is needed in upland rice system. The changing global climate makes dry season longer, resulting in the reduction of rice production. There should be an effort to introduce new variety of high-yielding and drought tolerant rice.  In this attempt, research was conducted to improve the genetic of Indica rice, particularly Gajahmungkur, Towuti and IR 64 varieties in order to find the somaclones with the characteristics above. As an approach, gamma-ray mutative induction was applied to be followed by selection in PEG. The regenerated shoot from the irradiated callus was then selected and acclimatisized in the greenhouse to obtain eighty three somaclones from the three varieties. PEG (molecular weight 6000) was applied to obtain the drought-tolerant somaclone. PEG was a selective agent used by which populations could be selected in a short time. Treatment with  20% PEG (equals to osmotic potential 1.2 Mpa) on the rice produced 16 somaclones from Gajahmungkur, 12 from Towuti and 18 from IR 64 putatively drought tolerant.   Key words : Oryza sativa, drought tolerance, PEG
Perbanyakan Cepat Jahe Merah melalui Teknik Kultur Jaringan Gati, Endang; Mariska, Ika
Buletin Penelitian Tanaman Rempah dan Obat Vol 3, No 1 (1988): Buletin Penelitian Tanaman Rempah dan Obat
Publisher : Pusat Penelitian dan Pengembangan Perkebunan

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Abstract

Investasi untuk pengadaan bibit hamper mencapai 40%. Metode pembiakan secara cepat pada jahe melalui kultur aringan telah dilakukan di Balai Penelitian Tanaman Rempah dan Obat. Eksplan yang digunakan diambil dari rimpang. Data menunjukkan bahwa medium yang mengandung kinetin tidak dapat menginduksi pembentukan tunas, sedangkan BAP 10 mg/l ditambah auksin 1 mg/l dapat menginduksi kalus kompak yang diikuti dengan 3-5 tunas adventif, dan terakhir terjadi induksi pembentukan akar. Kontaminasi bakteri dapat diatasi berturut-turut dengan menggunakan alcohol 70% selama 2 menit, HgCl2 0,5% selama 20 menit, Na-hipokhlorit 50% selama 10 menit dan akhirnya dilakukan pembilasan dengan air suling steril sebanyak 3 kali
Induksi Embriogenesis Somatik dari Jaringan Endosperma Jeruk Siam (Citrus nobilis Lour.) cv Simadu Kosmiatin, Mia; Purwito, Agus; Wattimena, Gustaff Adolf; Mariska, Ika
Jurnal Agronomi Indonesia (Indonesian Journal of Agronomy) Vol 42, No 1 (2014): JURNAL AGRONOMI INDONESIA
Publisher : Bogor Agricultural University

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ABSTRACTTriploid plants can be obtained from endosperm tissues through somatic embryogenesis regeneration. This research aimed to obtain somatic embryogenesis regeneration technique of tangerine endosperm. There were 3 experiments conducted in this research: 1) Embryogenic callus induction of tangerine endosperm. Endosperms isolated from fruits that were harvested from mother plants 11-13 weeks after anthesis and cultured on Murashige and Skoog (MS) medium by modified vitamin Morel and Wetmore (MW) which treated by 0.1 mg L-1 biotin, 500 mg L-1 malt extract (ME), 500 mg L-1 Casein hydrolisate (CH), 500 mg L-1 ME + 0.1 mg L-1 biotin, and 500 mg L-1CH + 0.1 mg L-1 biotin, 2) Maturation and germination of somatic embryos conducted by embryogenic callus cultured on MS medium by vitamin MW modified with addition of ABA, glutamine, and biotin, and 3) Plantlet elongation conducted on MS medium modified by MW vitamin with addition of GA3 and Kinetin. The best induction medium for embryogenic callus was modified MS enriched with 3 mg L-1 BA and 500 L-1 CH or ME, in succession 84.0 and 80.0%. The best medium for somatic embryos maturation with normal morphological plantlets (54.8%) was modified MS medium without plant growth regulator with higher rate of solidified agent (from 2.5 to 3 g L-1 Phytagel). Plantlets elongation was highly (0.9 cm) occurred on modified MS with enriched of 2.5 mg L-1 GA3. Keywords: Citrus nobilis (Lour.), endosperm culture, in vitro, Simadu tangerine
THE EFFECT OF PICLORAM AND LIGHT ON SOMATIC EMBRYOGENESIS REGENERATION OF PINEAPPLE Roostika, Ika; Khumaida, Nurul; Mariska, Ika; Wattimena, Gustaaf Adolf
Indonesian Journal of Agricultural Science Vol 13, No 2 (2012): October 2012
Publisher : Indonesian Agency for Agricultural Research and Development - MOA

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Smooth Cayenne is the largest pineapple type cultivated in Indonesia, but its vegetative planting materials for mass propagation are limited. Somatic embryogenesis is a potential method to be applied. The aim of this study was to investigate the somatic embryogenesis regeneration under the effect of picloram and light. Callus formation was induced by picloram (21, 41 and 62 μM) added with 9 μM thidiazuron. The calli were transferred onto MS or Bac medium  enriched with N-organic compounds with or without addition of 21 μM picloram under dark or light condition. The compact calli were subcultured onto MS medium supplemented with 4.65 μM kinetin, while the friable calli were  transferred onto BIG medium (modified MS + 1.1 μM benzyl adenine + 0.9 μM indole butyric acid + 0.09 μM giberelic acid) or B medium (MS + 0.018 mM benzyl adenine). The results showed that the events of somatic embryogenesis were started from cell polarization, asymmetrical division, proembryo formation as  embryogenic tissues and friable embryogenic tissues, and embryo development. The best treatment for callus induction was 21 μM picloram. The addition of 21 μM picloram on N-organic enriched medium and the use of light condition  proliferated embryogenic calli. The N-organic enriched Bac medium and light condition yielded the highest number of mature somatic embryos (17 embryos perexplant in 2 months). The B medium was better than BIG medium to develop  somatic embryos from friable embryogenic tissues. The somatic embryogenesis method presented is potential for pineapple mass propagation and artificial seedproduction.Abstrak Bahasa IndonesiaSmooth Cayenne merupakan kultivar nenas yang banyak dibudidayakan di  Indonesia, namun ketersediaan benih untuk perbanyakan massal masih terbatas. Embriogenesis somatikadalah metode yang potensial untuk produksi bibit secara massal. Tujuan penelitian adalah untuk mempelajari pengaruh pikloram dan pencahayaan terhadap regenerasi embriogenesis somatik nenas. Kalus diinduksi menggunakan pikloram (21, 41, dan 62 μM) dan penambahan thidiazuron 9 μM. Selanjutnya, kalus dipindahkan ke media MS atau Bac yang diperkaya dengansenyawa N-organik dengan atau tanpa penambahan pikloram 21 μM dalam kondisi gelap atau dengan pencahayaan. Kalus kompak disubkultur pada media MS yang mengandung kinetin 4,65 μM, sedangkan kalus remah dipindahkan ke media BIG (MS modifikasi + bensil adenin 1.1 μM + indole butyric acid 0,9 μM + giberelic acid 0,09 μM) atau media B (MS + bensil adenin 0,018 μM). Hasil penelitian  menunjukkan bahwa tahapan embriogenesis somatik diawali dengan polarisasi sel, pembelahan asimetris, pembentukan proembrio sebagai jaringan embriogenik danjaringan embriogenik remah, serta perkembangan embrio. Perlakuan terbaik untuk induksi kalus adalah pikloram 21 μM. Penambahan pikloram 21 μM pada media yang diperkaya dengan senyawa N-organik mampu meningkatkan jumlah kalusembriogenik. Media Bac yang diperkaya senyawa N-organik dan kondisi pencahayaan menghasilkan jumlah embrio somatik dewasa terbanyak (17 embrio per eksplan dalam 2 bulan). Media B lebih baik daripada media BIG untuk regenerasi embrio somatik dari jaringan embriogenik remah. Metode embriogenesis somatik yang dihasilkan dari penelitian ini berpotensi diterapkan untukperbanyakan massal dan produksi benih nenas.
Pembentukan Benih Sintetik Tanaman Nenas Roostika, Ika; Purnamaningsih, R; Supriati, Y; Mariska, Ika; Khumaida, N; Wattimena, GA
Jurnal Hortikultura Vol 22, No 4 (2012): Desember
Publisher : Indonesian Center for Horticultural Research and Development

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Abstract

Nenas merupakan tanaman buah tropis dan subtropis yang komersial. Kultivar Smooth Cayenne memiliki tipe dan jumlah propagul yang terbatas, sehingga diperlukan dukungan teknologi lainnya untuk produksi benih secara masal. Teknologi benih sintetik dapat diterapkan untuk produksi benih secara masal dan konservasi. Tujuan  penelitian ialah untuk mengetahui pengaruh kombinasi auksin dan sitokinin terhadap morfogenesis eksplan nenas yang terenkapsulasi, mengetahui pengaruh interaksi antara suhu penyimpanan dengan konsentrasi paklobutrazol atau manitol terhadap pertumbuhan eksplan nenas yang terenkapsulasi dan masa simpan. Penelitian dilaksanakan dari Bulan April sampai dengan Desember 2011 di Laboratorium Kultur Jaringan, Kelompok Peneliti Biologi Sel dan Jaringan, Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumberdaya Genetik Pertanian, Bogor. Percobaan disusun secara faktorial dalam rancangan acak lengkap terdiri atas enkapsulasi eksplan, pertumbuhan minimal menggunakan paklobutrazol, atau manitol yang dikombinasikan dengan suhu penyimpanan. Enkapsulasi dilakukan terhadap batang semu dan basal daun menggunakan Na-alginat 3% yang berisi media MS dengan penambahan BA (0, 1, 2, dan 3 mg/l) yang dikombinasikan dengan NAA (0, 1, 2, dan 3 mg/l). Untuk memacu proses diferensiasi, basal daun diberi praperlakuan menggunakan media MS yang mengandung BA 0,5 mg/l dan NAA 0,5 mg/l sebelum dienkapsulasi dengan perlakuan BA dan NAA pada konsentrasi 0; 0,5; dan 1 mg/l.  Pertumbuhan minimal dilakukan menggunakan paklobutrazol (0, 1, 2, dan 3 mg/l) atau manitol (0, 1, 2, 3, 4, dan 5%) pada suhu penyimpanan 15 dan 25 0C. Hasil penelitian menunjukkan bahwa basal daun nenas yang terenkapsulasi mampu berdiferensiasi setelah praperlakuan. Tidak terdapat interaksi yang nyata antara konsentrasi paklobutrazol dengan suhu penyimpanan terhadap daya hidup dan daya tembus kapsul tunas nenas. Biakan tersebut hanya dapat disimpan selama 1 bulan. Interaksi yang nyata juga tidak dijumpai antara konsentrasi manitol dengan suhu penyimpanan terhadap daya hidup dan daya tembus kapsul embrio somatik nenas. Manitol 4% mampu memperpanjang masa simpan hingga 4 bulan. Manitol dapat menggantikan aplikasi suhu rendah dalam penyimpanan kultur nenas yang terenkapsulasi. Pineapple is a commercial tropical and subtropical fruit crop. Smooth Cayenne cultivar has limited type and number of propagules so that it should be supported by the other technology to produce plenty seedlings. Artificial seed can be applied for seed production and conservation. The objectives of the study were to know the effect of combination treatments between auxin and cytokinin to the morphogenesis of encapsulated pineapple cultures, to know the effect of paclobutrazol, mannitol, and temperature of storage to the growth of encapsulated pineapple cultures. The experiment was conducted from April to December 2011 at Tissue Culture Laboratory, Researchers Group of Cell and Tissue of Biology, Indonesian Center for Agricultural Biotechnology and Genetic Resources Research and Development, Bogor. Factorial of a completey randomized design was used. The study consisted of encapsulation, minimal growth by using paclobutrazol or mannitol combined with storage temperature. Encapsulation was conducted by using 3% Na-alginat containing of MS medium with addition of BA (0, 1, 2, and 3 mg/l) combined with NAA (0, 1, 2, and 3 mg/l). To promote differentiation, leaf bases were pre-cultured on MS media containing BA and NAA at concentration of 0.5 mg/l respectively prior to encapsulated by BA and NAA (0; 0.5; and 1 mg/l). Minimal growth was conducted by using paclobutrazol (0, 1, 2, and 3 mg/l), or mannitol (0, 1, 2, 3, 4, and 5%), and combined with storage temperature (15 and 25 0C). The results showed that encapsulated leaf bases of pineapple could differentiate after pre-treatment. There was no interaction between paclobutrazol and temperature to the survival rate and emergence rate of the encapsulated cultures. The encapsulated shoots could be stored for 1 months. There was also no interaction between mannitol and temperature to the survival rate and emergence rate of the encapsulated cultures. By using somatic embryos and 4% mannitol, the storage period could be prolonged for 4 months. Mannitol could substitute the use of low temperature in the conservation of encapsulated pineapple cultures.
Regenerasi Tanaman Sedap Malam Melalui Organogenesis dan Embriogenesis Somatik Rostika, Ika; Mariska, Ika; Purnamaningsih, R
Jurnal Hortikultura Vol 15, No 4 (2005): Desember 2005
Publisher : Indonesian Center for Horticultural Research and Development

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Abstract

Secara konvensional perbanyakan tanaman sedap malam dilakukan melalui umbi. Semakin kecil ukuran umbi semakin lama tanaman berbunga. Penerapan teknik kultur in vitro diharapkan dapat membantu perbanyakan tanaman secara masal. Hingga saat ini, teknik kultur in vitro tanaman sedap malam belum pernah dilaporkan di Indonesia. Penelitian ini bertujuan memperoleh formulasi media yang efektif menginduksi organogenesis dan embriogenesis kultur in vitro tanaman sedap malam serta memacu regenerasinya. Percobaan dibagi menjadi 4 tahap, yaitu (1) induksi tunas, (2) multiplikasi tunas, (3) induksi kalus embriogenik, dan (4) regenerasi kalus embriogenik. Media induksi tunas yang diuji adalah MS+BA 0 ppm, MS+BA 3 ppm, MS+BA 5 ppm, dan MS+BA 7 ppm. Pemacuan multiplikasi tunas lanjut dilakukan pada media subkultur MS+BA 7 ppm+glutamin 100 ppm, MS+BA 7 ppm, DKW+TDZ 7 ppm, dan DKW+TDZ 7 ppm+glutamin 100 ppm. Untuk induksi kalus embriogenik, media induksi kalus yang diujikan adalah MS+2,4-D 2,5 ppm, MS +2,4-D 5 ppm, dan MS+2,4-D 10 ppm. Untuk meregenerasikan kalus embriogenik, media yang diujikan MS+BA 2 ppm+TDZ 0,2 ppm, MS+BA 3 ppm+TDZ 0,4 ppm, MS+zeatin 1ppm+kinetin 1ppm, dan MS+zeatin 0,5 ppm+kinetin 2 ppm. Hasil percobaan menunjukkan bahwa pembentukan tunas terbanyak diperoleh dari media BA 3 ppm (80%) namun inisiasi tunas tercepat dihasilkan pada media BA 0 ppm. Formula media MS+BA 7 ppm+glutamin 100 ppm menghasilkan jumlah tunas dan akar terbanyak. Penggunaan MS+2,4-D 5 ppm dapat menginduksi kalus embriogenik dengan persentase pembentukan nodul sebesar 18,75% dan jumlah nodul yang terbentuk sebanyak 3,6 dengan visual kalus yang paling baik. Setelah disubkultur, calon tunas terbanyak (17) dihasilkan dari perlakuan MS+BA 2 ppm+TDZ 0,4 ppm. Kalus embriogenik pada media MS+zeatin 0,5 ppm+kinetin 2 ppm dapat berkembang membentuk benih somatik.Regeneration of tuberose through organogenesis and embryogenesis. Tuberose is normally propagated by the tuber. The smaller size of tuber the longer time plant to flower. The application of in vitro culture technique might be used for mass propagation. Up to know, the research of in vitro culture of tuberose in Indonesia has not been reported. The objective of the study was to find out media formulation for organogenesis and embryogenesis. The experiments consisted of 4 steps of (1) shoot induction, (2) shoot multiplication, (3) induction of embryogenic callus, and (4) regeneration of embryogenic callus. The treatments for shoot induction were MS+BA 0 ppm, MS+BA 3 ppm, MS+BA 5 ppm, and MS+BA 7 ppm. The shoots were multiplied on media MS+BA 7ppm+glutamine 100ppm, MS+BA 7 ppm, DKW+TDZ 7 ppm, and DKW+TDZ 7 ppm+glutamin 100 ppm. For induction of embryogenic callus, the treatments were MS+2.4-D 2.5 ppm, MS+2,4-D 5 ppm, and MS+2.4-D 10 ppm. For regeneration of embryogenic callus, the treatments were MS+BA 2 ppm+TDZ 0.2 ppm, MS+BA 3 ppm +TDZ 0.4 ppm, MS+zeatin 1ppm+kinetin 1ppm, and MS+zeatin 0.5 ppm+kinetin 2 ppm. The results showed that the highest shoot formation was obtained from media MS+BA 3 ppm but the earliest shoot initiation was obtained from media MS+BA 0 ppm. The media formulation of MS+BA 7 ppm+glutamine 100 ppm gave the highest number of shoot and root. The application of media MS+2.4-D 5 ppm could induce embryogenic callus with high percentage of nodul formation (18.75%) and high number of nodul (3.6) with the best visual calli. After subculturing, the highest number of nodul (17) was obtained from media MS+BA 2 ppm+TDZ 0.4 ppm. The embryogenic callus from media MS+zeatin 0.5 ppm+kinetin 2 ppm could develop to form somatic seed.