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All Journal Indonesian Journal of Agricultural Science Jurnal Tanaman Industri dan Penyegar Journal of Tropical Crop Science Buletin Penelitian Tanaman Rempah dan Obat
Meynarti Sari Dewi Ibrahim
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DIRECT AND INDIRECT SOMATIC EMBRYOGENESIS ON ARABICA COFFEE (Coffea arabica) Ibrahim, Meynarti Sari Dewi; Hartati, Rr. Sri; Rubiyo, Rubiyo; Purwito, Agus; Sudarsono, Sudarsono
Indonesian Journal of Agricultural Science Vol 14, No 2 (2013): October 2013
Publisher : Indonesian Agency for Agricultural Research and Development - MOA

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Abstract

Propagation of Coffea arabica L. through direct and indirect somatic embryogenesis technique is promising for producing large number of coffee seedlings. The objectives of the research were to evaluate methods for direct and indirect somatic embryo-genesis induction of C. arabica var. Kartika. The explants were the youngest fully expanded leaves of arabica coffee. The evalu-ated medium was modified Murashige and Skoog (MS) medium supplemented with a combination of 2.26 µM 2,4-D + 4.54 or 9.08 µM thidiazuron; 4.52 µM 2,4-D + 4.54 or 9.08 µM thidiazuron; or 9.04 µM 2,4-D + 9.08 µM thidiazuron. Both calli (100 mg) and pre-embryos developed from the edge of leaf explants were subcultured into regeneration medium (half strength MS with modified vitamin, supplemented with kinetine 9.30 µM and adenine sulfate 40 mg L-1). The results showed coffee leaf explant cultured on medium containing 2.26 µM 2,4-D + 4.54 or 9.08 µM thidiazuron to induce direct somatic embriogenesis from explant, while that of 4.52 or 9.04 µM 2,4-D + 9.08 µM thidiazuron to induced indirect somatic embrio-genesis. The medium for calli induction from coffee by explants was medium supplemented with 4.52 or 9.04 µM 2,4-D in combination with 9.08 µM thidiazuron. On the other hand, the best medium for activation of induction of somatic embryos was MS medium supplemented with 9.04 µM 2,4-D + 9.08 µM thidiazuron. Based on this results, the first step for developing micropropagation for coffee has been resolved. The subsequent studies will be directed to evaluate agronomic performance of the derived planting materials.
Pengaruh Komposisi Media terhadap Pembentukan Kalus Embriogenesis Somatik Kopi Arabika ( Coffea arabica ) Ibrahim, Meynarti Sari dewi; Sudarsono, Sudarsono; Rubiyo, Rubiyo; Syafaruddin, Syafaruddin
Jurnal Tanaman Industri dan Penyegar Vol 3, No 1 (2012): Buletin Riset Tanaman Rempah Dan Aneka Tanaman Industri
Publisher : Pusat Penelitian dan Pengembangan Perkebunan

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Abstract

Induksi embrio somatik pada kopi arabika (Coffea arabica) dengan menggunakan beberapa zat pengatur tumbuh (ZPT) telah berhasil dilakukan. Pengaruh komposisi media terutama kombinasi antara jenis ZPT yang berbeda dan tanggap genotipe tanaman dilaporkan sangat bervariasi. Tujuan penelitian untuk mengkaji pengaruh pemberian 2,4-D dan kinetin dalam proses pembentukan dan pertumbuhan kalus embriogenik asal daun. Penelitian dilakukan di Laboratorium Pusat Penelitian dan Pengembangan Perkebunan (Puslitbangbun) Agustus 2011 sampai Januari 2012. Bahan tanaman yang digunakan adalah daun dari kopi arabika varietas Sigarar Utang yang merupakan tanaman koleksi Balai Penelitian Tanaman Industri dan Penyegar (Balittri). Penelitian menggunakan rancangan acak lengkap dengan 5 ulangan, masing-masing ulangan terdiri dari 5 eksplan. Peubah yang diamati, meliputi persentasi kalus yang terbentuk, morfologi kalus, berat basah kalus, dan jumlah globular. Hasil menunjukkan semua perlakuan dapat membentuk kalus, pertambahan berat eksplan tertinggi diperoleh pada media kombinasi 2,4-D 1 mg/l atau 2 mg/l dan kinetin 1 sampai 4 mg/l. Embrio somatik terbanyak diperoleh pada media yang diberi 2,4-D 0,5 mg/l dan kinetin 1 mg/l. Selain kalus, massa proembrio dan embrio, juga terbentuk akar adventif yang jumlahnya tidak nyata antar perlakuan.  The Effect of Composition Media to Callus Formation of Somaticembryogenesis of Arabica Coffee (Coffea arabica)ABSTRACT Induction of somatic embryos with plant growth regulators (PGR) has successfully performed in arabica coffee. However, the influence of media composition combined with different PGR, explants and genotype of plants is widely various in response yields. The objective of this study was to examine the effects of 2,4-D and kinetine in process of formation and growth of embryogenic callus developed from leaves of arabica coffee. The studiy was carried out at a laboratory of Indonesian Research Center for Estate Crops (Puslitbangbun) from August 2011 to January 2012. Plant materials used are coffee leaves var. Sigarar Utang taken from a germplasm collection of the crop grown at Pakuwon Research Station, Indonesian Research Institute for Industry Crops (Balittri) located at Sukabumi, West Java. A completely randomized design with 5 replications and plot size of five explants was used. Parameters observed are percentage of callus formation, morphology of the callus, fresh weight of callus, and number of globular. Results show that all treatments examined are able to form callus. The highest increase in weight of explants was obtained from the media treated with 2,4-D (conc. of 1mg/l or 2 mg/l) and kinetin (conc. of 1 to 4 mg/l). While, the most number of somatic embryo formed was obtained from those of treated with 2,4-D 0.5 mg/l and kinetin 1 mg/l. In addition to callus formation, proembryo mass, embryo and adventive roots were also formed in spite of not significant between different the treatments.
Penggunaan Air Kelapa dan Beberapa Auksin untuk Induksi Multiplikasi Tunas dan Perakaran Lada Secara In Vitro Sulistiyorini, Indah; Ibrahim, Meynarti Sari Dewi; Syafaruddin, Syafaruddin
Jurnal Tanaman Industri dan Penyegar Vol 3, No 3 (2012): Buletin Riset Tanaman Rempah Dan Aneka Tanaman Industri
Publisher : Pusat Penelitian dan Pengembangan Perkebunan

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Abstract

Peningkatan produktivitas lada perlu didukung oleh ketersediaan benih unggul. Perbanyakan lada secara in vitro dapat digunakan sebagai alternatif untuk menghasilkan benih lada dalam jumlah banyak dan waktu yang relatif singkat. Salah satu faktor yang menentukan keberhasilan kultur in vitro adalah penggunaan zat pengatur tumbuh. Penelitian dilakukan di Laboratorium Kultur Jaringan Balai Penelitian Tanaman Industri dan Penyegar mulai bulan Maret-September 2011. Penelitian terdiri dari 2 kegiatan yaitu induksi multiplikasi tunas dan induksi perakaran. Masing-masing bertujuan untuk menganalisis penggunaan konsentrasi air kelapa terhadap multiplikasi tunas lada dan pengaruh penggunaan jenis dan konsentrasi auksin terhadap induksi perakaran lada secara in vitro. Rancangan yang digunakan adalah rancangan acak lengkap dengan uji lanjut DMRT. Perlakuan induksi multiplikasi terdiri dari konsentrasi air kelapa, yaitu 10, 20, 30, 40, 50% dan sebagai pembanding adalah BA 0,3 mg/l, sedangkan induksi perakaran lada digunakan beberapa auksin, yaitu IBA, IAA dan 2,4-D dengan konsentrasi masing-masing adalah 0,1, 0,3, dan 0,5 mg/l. Hasil penelitian menunjukkan penggunaan air kelapa untuk perlakuan induksi multiplikasi tunas pada semua konsentrasi lebih memacu pembentukan akar, selain itu kultur yang dihasilkan mempunyai pertumbuhan normal dan lebih vigor dibandingkan perlakuan BA 0,3 mg/l. Perlakuan BA 0,3 mg/l menghasilkan jumlah tunas dan jumlah daun lebih banyak dibandingkan perlakuan air kelapa sebesar 2,69 dan 10,73. Penggunaan IAA 0,1 mg/l untuk induksi perakaran mampu menginduksi akar sebanyak 8,26 lebih banyak dibandingkan auksin yang lain.  The Use of Coconut Water And Several Auxin for Shoot Multiplication And Rooting Induction in Black Pepper In VitroABSTRACT Increased productivity of pepper should be supported by the availability of improved seed. Propagation black pepper in vitro can be used as an alternative to produce large amounts of black pepper cuttings in a relatively short time. One of the factors that determine the success of in vitro culture is the use of plant growth regulators used. Research was conducted in the laboratory tissue culture from March to September 2011. This research consists of two activities, the induction of shoot multiplication and rooting induction. Each aims to analyze the addition of coconut water concentration on shoot multiplication black pepper and determine the effect of the addition of the type and concentration of auxin for induction in vitro rooting of black pepper. Design used were completely randomized design and use advanced testing DMRT. Treatment consisted of induction multiplication coconut water concentration, namely 10, 20, 30, 40, 50%, and as a comparison is BA 0.3 mg/l, and black pepper root induction treatment using several auxin is IBA, IAA and 2.4-D with the concentration of each was 0.1 mg/l, 0.3 mg/l and 0.5 mg/l.  The results showed the use of coconut water for shoot multiplication induction treatment at all concentrations stimulate root formation, in addition to the culture that has produced more normal growth and vigor than the treatment of BA 0.3 mg/l. Treatment BA 0.3 mg/l produce shoots leaves more than coconut water treatment at 2.69 and 10.73. The use of IAA 0.1 mg/l for induction were able to induce root 8.26 more as compared to other auxin.
Induksi Kalus Embriogenik dan Daya Regenerasi Kopi Arabika Menggunakan 2,4- Dichlorophenoxyacetic Acid dan 6-Benzyladenine Ibrahim, Meynarti Sari Dewi; Hartati, Rr Sri; Rubiyo, Rubiyo; Purwito, Agus; Sudarsono, Sudarsono
Jurnal Tanaman Industri dan Penyegar Vol 4, No 2 (2013): Buletin Riset Tanaman Rempah dan Aneka Tanaman Industri
Publisher : Pusat Penelitian dan Pengembangan Perkebunan

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Abstract

Embriogenesis somatik kopi Arabika (Coffea arabica L.) masih mengalami kendala dalam meregenerasikan planlet dari eksplan yang dikulturkan. Kemampuan eksplan daun membentuk embrio dalam proses embriogenesis somatik kopi sangat dipengaruhi oleh komposisi media dan zat pengatur tumbuh. Penelitian bertujuan untuk melihat pengaruh pemberian 2,4-Dichlorophenoxyacetic Acid dan 6-Benzyladenine dalam proses pembentukan kalus embriogenik dan daya regenerasi kopi Arabika. Penelitian dilakukan di Unit Pengembangan Benih Unggul Pertanian, Badan Penelitian dan Pengembangan Pertanian pada bulan Juli 2011 sampai Desember 2012. Bahan tanaman yang digunakan adalah daun kopi Arabika varietas S795 koleksi Balai Penelitian Tanaman Industri dan Penyegar. Rancangan perlakuan menggunakan rancangan acak lengkap dengan 6 ulangan, masing-masing ulangan terdiri dari 5 eksplan. Induksi kalus menggunakan 5 kombinasi perlakuan 2,4-D 1 mg/l + BA 0 mg/l; 2,4-D 1 mg/l + BA 1 mg/l; 2,4-D 1 mg/l + BA 2 mg/l; 2,4-D 2 mg/l + BA 1 mg/l; dan kontrol (tanpa penambahan 2,4-D dan BA). Peubah yang diamati meliputi jumlah kalus, persentasi kalus embriogenik, berat basah kalus, dan jumlah proembrio. Hasil penelitian menunjukkan perlakuan 2,4-D 1 mg/l + BA 0 mg/l; 2,4-D 1 mg/l + BA 1 mg/l; 2,4-D 1 mg/l + BA 2 mg/l; dan 2,4-D 2 mg/l + BA 1 mg/l dapat membentuk kalus kecuali perlakuan kontrol. Berat kalus, persentasi kalus embriogenik, dan jumlah proembrio tertinggi diperoleh pada media kombinasi 2,4-D 2 mg/l dan BA 1 mg/l. Kalus yang mampu beregenerasi berasal dari media kombinasi 2,4-D 1 mg/l dan BA 2 mg/l dengan persentasi 16,67% dengan 6 kecambah per 0,2 gram kalus.Kata Kunci: Coffea arabica, 2,4-Dichlorophenoxyacetic Acid, 6-Benzyladenine, embriogenesis somatikRegeneration of planlets from cultured explants has been an obstacle in somatic embryogenesis of arabica coffee (Coffea arabica L.). The ability of leaf explants to generate embryos in somatic embryogenesis process of coffee was affected by composition of media and plant growth regulators. The objectives of the research was to examine the effect of 2,4-Dichlorophenoxyacetic Acid dan 6-Benzyladenine in the process of embryogenic callus and regeneration potential of arabica coffee. The study was conducted at Agricultural Superior Seed Development Unit, Indonesian Agency for Agricultural Research and Development (IAARD) from July 2011 to December 2012. Plant material used was leaves of S795 variety which is collected by Indonesian Industrial and Beverage Crops Research Institute. The research was arranged in completely randomized design with 6 replications, each replication consist of 5 explants. Callus induction used 5 treatments, i.e. 2,4-D 1 mg/l + BA 0 mg/l; 2,4-D 1 mg/l + BA 1 mg/l; 2,4-D 1 mg/l + BA 2 mg/l; 2,4-D 2 mg/l + BA 1 mg/l; and control (without 2,4-D and BA). Variables observed were number of callus, percentage of embryogenic callus, callus fresh weight and number of proembryo. Result showed that all treatments can produce the callus except control. Combination of 2,4-D 2 mg/l and BA 1 mg/l gave the highest of fresh weight of callus, percentage of embryogenic callus, and number of proembryo. Regenerating callus of 16.67% with the number of sprouts of 6 per 0.2 gram only derived from combination of 2,4-D 1 mg/l BA and 2 mg/l. 
Pengaruh Komposisi Media terhadap Pembentukan Kalus Embriogenesis Somatik Kopi Arabika ( Coffea arabica ) Ibrahim, Meynarti Sari dewi; Sudarsono, Sudarsono; Rubiyo, Rubiyo; Syafaruddin, Syafaruddin
Jurnal Tanaman Industri dan Penyegar Vol 3, No 1 (2012): Buletin Riset Tanaman Rempah Dan Aneka Tanaman Industri
Publisher : Pusat Penelitian dan Pengembangan Perkebunan

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Abstract

Induksi embrio somatik pada kopi arabika (Coffea arabica) dengan menggunakan beberapa zat pengatur tumbuh (ZPT) telah berhasil dilakukan. Pengaruh komposisi media terutama kombinasi antara jenis ZPT yang berbeda dan tanggap genotipe tanaman dilaporkan sangat bervariasi. Tujuan penelitian untuk mengkaji pengaruh pemberian 2,4-D dan kinetin dalam proses pembentukan dan pertumbuhan kalus embriogenik asal daun. Penelitian dilakukan di Laboratorium Pusat Penelitian dan Pengembangan Perkebunan (Puslitbangbun) Agustus 2011 sampai Januari 2012. Bahan tanaman yang digunakan adalah daun dari kopi arabika varietas Sigarar Utang yang merupakan tanaman koleksi Balai Penelitian Tanaman Industri dan Penyegar (Balittri). Penelitian menggunakan rancangan acak lengkap dengan 5 ulangan, masing-masing ulangan terdiri dari 5 eksplan. Peubah yang diamati, meliputi persentasi kalus yang terbentuk, morfologi kalus, berat basah kalus, dan jumlah globular. Hasil menunjukkan semua perlakuan dapat membentuk kalus, pertambahan berat eksplan tertinggi diperoleh pada media kombinasi 2,4-D 1 mg/l atau 2 mg/l dan kinetin 1 sampai 4 mg/l. Embrio somatik terbanyak diperoleh pada media yang diberi 2,4-D 0,5 mg/l dan kinetin 1 mg/l. Selain kalus, massa proembrio dan embrio, juga terbentuk akar adventif yang jumlahnya tidak nyata antar perlakuan.  The Effect of Composition Media to Callus Formation of Somaticembryogenesis of Arabica Coffee (Coffea arabica)ABSTRACT Induction of somatic embryos with plant growth regulators (PGR) has successfully performed in arabica coffee. However, the influence of media composition combined with different PGR, explants and genotype of plants is widely various in response yields. The objective of this study was to examine the effects of 2,4-D and kinetine in process of formation and growth of embryogenic callus developed from leaves of arabica coffee. The studiy was carried out at a laboratory of Indonesian Research Center for Estate Crops (Puslitbangbun) from August 2011 to January 2012. Plant materials used are coffee leaves var. Sigarar Utang taken from a germplasm collection of the crop grown at Pakuwon Research Station, Indonesian Research Institute for Industry Crops (Balittri) located at Sukabumi, West Java. A completely randomized design with 5 replications and plot size of five explants was used. Parameters observed are percentage of callus formation, morphology of the callus, fresh weight of callus, and number of globular. Results show that all treatments examined are able to form callus. The highest increase in weight of explants was obtained from the media treated with 2,4-D (conc. of 1mg/l or 2 mg/l) and kinetin (conc. of 1 to 4 mg/l). While, the most number of somatic embryo formed was obtained from those of treated with 2,4-D 0.5 mg/l and kinetin 1 mg/l. In addition to callus formation, proembryo mass, embryo and adventive roots were also formed in spite of not significant between different the treatments.
Penggunaan Air Kelapa dan Beberapa Auksin untuk Induksi Multiplikasi Tunas dan Perakaran Lada Secara In Vitro Sulistiyorini, Indah; Ibrahim, Meynarti Sari Dewi; Syafaruddin, Syafaruddin
Jurnal Tanaman Industri dan Penyegar Vol 3, No 3 (2012): Buletin Riset Tanaman Rempah Dan Aneka Tanaman Industri
Publisher : Pusat Penelitian dan Pengembangan Perkebunan

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Abstract

Peningkatan produktivitas lada perlu didukung oleh ketersediaan benih unggul. Perbanyakan lada secara in vitro dapat digunakan sebagai alternatif untuk menghasilkan benih lada dalam jumlah banyak dan waktu yang relatif singkat. Salah satu faktor yang menentukan keberhasilan kultur in vitro adalah penggunaan zat pengatur tumbuh. Penelitian dilakukan di Laboratorium Kultur Jaringan Balai Penelitian Tanaman Industri dan Penyegar mulai bulan Maret-September 2011. Penelitian terdiri dari 2 kegiatan yaitu induksi multiplikasi tunas dan induksi perakaran. Masing-masing bertujuan untuk menganalisis penggunaan konsentrasi air kelapa terhadap multiplikasi tunas lada dan pengaruh penggunaan jenis dan konsentrasi auksin terhadap induksi perakaran lada secara in vitro. Rancangan yang digunakan adalah rancangan acak lengkap dengan uji lanjut DMRT. Perlakuan induksi multiplikasi terdiri dari konsentrasi air kelapa, yaitu 10, 20, 30, 40, 50% dan sebagai pembanding adalah BA 0,3 mg/l, sedangkan induksi perakaran lada digunakan beberapa auksin, yaitu IBA, IAA dan 2,4-D dengan konsentrasi masing-masing adalah 0,1, 0,3, dan 0,5 mg/l. Hasil penelitian menunjukkan penggunaan air kelapa untuk perlakuan induksi multiplikasi tunas pada semua konsentrasi lebih memacu pembentukan akar, selain itu kultur yang dihasilkan mempunyai pertumbuhan normal dan lebih vigor dibandingkan perlakuan BA 0,3 mg/l. Perlakuan BA 0,3 mg/l menghasilkan jumlah tunas dan jumlah daun lebih banyak dibandingkan perlakuan air kelapa sebesar 2,69 dan 10,73. Penggunaan IAA 0,1 mg/l untuk induksi perakaran mampu menginduksi akar sebanyak 8,26 lebih banyak dibandingkan auksin yang lain.  The Use of Coconut Water And Several Auxin for Shoot Multiplication And Rooting Induction in Black Pepper In VitroABSTRACT Increased productivity of pepper should be supported by the availability of improved seed. Propagation black pepper in vitro can be used as an alternative to produce large amounts of black pepper cuttings in a relatively short time. One of the factors that determine the success of in vitro culture is the use of plant growth regulators used. Research was conducted in the laboratory tissue culture from March to September 2011. This research consists of two activities, the induction of shoot multiplication and rooting induction. Each aims to analyze the addition of coconut water concentration on shoot multiplication black pepper and determine the effect of the addition of the type and concentration of auxin for induction in vitro rooting of black pepper. Design used were completely randomized design and use advanced testing DMRT. Treatment consisted of induction multiplication coconut water concentration, namely 10, 20, 30, 40, 50%, and as a comparison is BA 0.3 mg/l, and black pepper root induction treatment using several auxin is IBA, IAA and 2.4-D with the concentration of each was 0.1 mg/l, 0.3 mg/l and 0.5 mg/l.  The results showed the use of coconut water for shoot multiplication induction treatment at all concentrations stimulate root formation, in addition to the culture that has produced more normal growth and vigor than the treatment of BA 0.3 mg/l. Treatment BA 0.3 mg/l produce shoots leaves more than coconut water treatment at 2.69 and 10.73. The use of IAA 0.1 mg/l for induction were able to induce root 8.26 more as compared to other auxin.
The Induction of Primary and Secondary Somatic Embryo to Support Arabica Coffee Propagation Ibrahim, Meynarti Sari Dewi; Hartati, Raden Roro Sri; Rubiyo, Rubiyo; Purwito, Agus; Sudarsono, Sudarsono
Journal of Tropical Crop Science Vol 2 No 3 (2015): Journal of Tropical Crop Science
Publisher : Department of Agronomy and Horticulture, IPB University

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (299.45 KB) | DOI: 10.29244/jtcs.2.3.6-13

Abstract

The primary and secondary somatic embryogenesis can be used to propagate Coffea arabica L clonally.  However, the success of this propagation was depended on plant growth regulator and varieties. This study aimed to examine the possibility of 2,4-D and thidiazuron application to form primary and secondary somatic embryo to support Arabica coffee clonal propagation. The study consisted of two activities (1) 2,4-D and thidiazuron Application to Induce Primary Somatic Embryogenesis of Arabica Coffee and (2) The Application of thidiazuron in Solid and Semi-Solid Media to Induce Secondary Somatic Embryos.  The results indicated significant effect of varieties and plant growth regulator on fresh weight, number of torpedo and germinated embryo.  However, it showed no significant effect on callus formation percentage. The best medium to induce primary somatic embryogenesis depending on variety, on the treatment of 4.52 μM 2,4 -D +18.16 μM thidiazuron was the best for AS2K and Sigarar Utang varieties, S 795 at 4.52 μM 2,4-D + 9.08 μM thidiazuron, whereas Kartika at 4.52 μM 2.4-D + 13.62 μM thidiazuron.  The morphology of coffee somatic embryo was normal.  Primary somatic embryo was developed indirectly, whereas the secondary somatic embryo was directly.  The application of 9.08 μM thidiazuron  increased the percentage and number of secondary somatic embryos, hence enhancing number of Arabica coffee planlet. Keywords : Coffea arabica L, 2,4-D, thidiazuron, semi-solid media, Indirect somatic embryogenesis
STUDI PENDAHULUAN : INDUKSI KALUS EMBRIOGENIK DARI EKSPLAN DAUN Echinaceae purpurea Ibrahim, Meynarti Sari Dewi; Kristina, N. Nova; Bermawie, Nurliani
Buletin Penelitian Tanaman Rempah dan Obat Vol 15, No 2 (2004): BULETIN PENELITIAN TANAMAN REMPAH DAN OBAT
Publisher : Pusat Penelitian dan Pengembangan Perkebunan

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/bullittro.v15n2.2004.%p

Abstract

Echinaceae is an introduced medicinal plant used to improve immune system of the body. Lately, interest on Echinaceae increased, however, good plant material for development is limited. One method to multiply plant material is the in vitro culture, so that research and development especially in vitro multiplication is required. The aim of the experiment is to procure a medium for the induction of embryonic callus and the technique of regeneration. The experiment is conducted at the Laboratory of Breeding and Genetic Resources, ISMECRI from January to December 2002, consisted of 2 steps, namely(1). Embryonic callus induction, (2). Callus regeneration. In the first step, explants were transferred into MS medium enrich with BA 0,1 mg/l + 2,4 D 0,5 mg/l and a combination medium of MS and LS medium supplemented with glutamine + BA (0,1 mg/l; 0,2 mg/l) + 2,4 D (0,5mg/l ; 1 mg/l). In the second step, regeneration was conducted on medium MS combined with LS + BA (0 mg/l + 0,2 mg/l + 0,4 mg/l), MS combined with LS + Kinetine (0,2 mg/l; 0,4 mg/l). The results showed that embryonic callus was obtained from culture of leaf explants on MS medium combined with LS. The best treatment for step regeneration was MS combined LS + BA 0,4 mg/l for shoot formation while MS combined LS + kinetine 0,4 mg/l induced root formation. 
Co-Authors Agus Purwito Hartati, Raden Roro Sri Indah Sulistiyorini Kristina, N. Nova Nurliani Bermawie Rr Sri Hartati, Rr Sri Rr. Sri Hartati Rubiyo Rubiyo Sudarsono Syafaruddin Syafaruddin