Pengaruh TDZ terhadap induksi embrio somatik sagu (Metroxylon  sagu Rottb.) pada tiga metode kultur berbeda (Effect of TDZ on the somatic embryo induction of sago palm (Metroxylon  sagu Rottb.) in three different culture methods)

AbstractA right combination of cytokinin is able to support the process of callus differentiation to somatic embryo formation in plant somatic embryogenesis. Liquid culture application could increase the efficiency of in vitro culture process on plants. This research aimed to determine the best concentration of TDZ combined with kinetin for callus differentiation to somatic embryo of sago palm on three culture methods. Plant material used was embryogenic callus derived from tips meristem culture from sucker of Alitir sago palm. Callus was cultured on modified MS media added with: 0.0, 0.1, 0.5 and 1.0 mg/L TDZ combined with 0.5 mg/L kinetin for 12 weeks with subcultures every 6 weeks. Three culture methods used were suspension, temporary immersion system (TIS), and solid media. There were 12 treatments with 4 replicates. The results showed that the highest number of somatic embryos was achieved on TIS culture with 1.0 mg/L TDZ and 0.5 mg/L kinetin in 6 weeks (167.3 embryos/flask) and 12 weeks (389.2 embryos/flask) with its fresh weight of 18.4 g and 29.1 g, respectively. The highset survival rate in final culture (12 weeks) was achieved on TIS culture with 1.0 mg/L TDZ and 0.5 mg/L kinetin (100%). The shortest time for somatic embryos expression was achieved on TIS culture with 1.0 mg/L TDZ and 0.5 mg/L kinetin in two weeks after culture. Histological analysis of early-stage somatic embryos showed the presence of dense and compact cellular arrangements which formed growth spot axis for shoot or SAM (shoot apical meristem) and root or RAM (root apical meristem) that connected each other. [Key words: culture method, embryogenic callus, Metroxylon sagu Rottb., kinetin, sago palm, TDZ] AbstrakAplikasi kombinasi sitokinin yang tepat dapat mendorong proses diferensiasi kalus membentuk embrio somatik pada proses embriogenesis somatik tanaman. Penggunaan metode kultur cair dapat meningkatkan efisiensi proses kultur in vitro tanaman. Penelitian ini bertujuan untuk menentukan konsentrasi TDZ terbaik dikombinasikan dengan kinetin dalam proses diferensiasi kalus membentuk embrio somatik tanaman sagu pada tiga metode kultur. Bahan tanam penelitian berupa kalus embriogenik tanaman sagu asal kultur meristem pucuk dari anakan sagu jenis Alitir. Kalus dikulturkan pada media modifikasi dengan penambahan TDZ dengan konsentrasi 0,1; 0,5; dan 1,0 mg/L dikombinasikan dengan kinetin 0,5 mg/L selama 12 minggu yang disubkultur pada umur 6 minggu. Metode kultur yang digunakan terdiri atas tiga macam yaitu: kultur suspensi, sistem perendaman sesaat (SPS) dan media padat. Perlakuan terdiri atas 12 kombinasi perlakuan dengan empat ulangan. Hasil penelitian menunjukkan bahwa rerata jumlah embrio somatik tertinggi dicapai pada perlakuan metode kultur SPS dengan TDZ 1,0 mg/L baik pada umur kultur 6 minggu (167,3 buah) maupun umur 12 minggu (389,2 buah). Rerata bobot segar tertinggi juga diperoleh pada perlakuan metode kultur SPS dengan TDZ 1,0 mg/L pada umur kultur 6 minggu (18,4 g) dan 12 minggu (29,1 g). Rerata daya hidup kultur akhir (12 minggu) tertinggi sebesar 100% diperoleh pada perlakuan SPS. Induksi embrio somatik tercepat yakni setelah dua minggu diperoleh pada metode kultur SPS dengan TDZ 1,0 mg/L dikombinasikan dengan kinetin 0,5 mg/L. Analisis histologi embrio somatik stadium awal menunjukkan adanya susunan sel yang rapat dan kompak yang menyusun semacam poros atau berkas titik tumbuh tunas atau SAM (shoot apical meristem) maupun akar atau RAM (root apical mersitem) yang saling terhubung.[Kata kunci: kalus embriogenik, metode kultur, kinetin, TDZ, sagu, Metroxylon sagu]

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EFFECT OF GAMMA IRRADIATION ON THE GROWTH AND DEVELOPMENT OF SAGO PALM (Metroxylon sagu Rottb.) CALLI

Indonesian Journal of Agricultural Science ◽

10.21082/ijas.v17n1.2016.p35-40 ◽

2017 ◽

Vol 17 (1) ◽

pp. 35

Author(s):

Imron Riyadi ◽

Sumaryono Sumaryono

Keyword(s):

Gamma Irradiation ◽

Somatic Embryo ◽

Somatic Embryos ◽

Growth And Development ◽

Gamma Ray ◽

Embryogenic Calli ◽

Material Source ◽

Plant Materials ◽

Sago Palm ◽

Metroxylon Sagu

The application of gamma irradiation on plant materials may increase the genetic variation of the offspring with useful traits. The experiment was conducted to determine the effect of irradiation dosage of gamma ray on growth and development of sago palm (Metroxylon sagu) calli. Friable calli of sago palm derived from suspension culture were used as a material source. The primary calli were initiated from apical meristematic tissues of sago palm suckers of Alitir variety from Merauke, Papua. The treatments used were dosage of gamma ray irradiation at 0, 5, 10, 15, 20 and 25 Gy. The treated calli were then subcultured on modified Murashige and Skoog (MMS) solid medium containing 3% sucrose and 0.1% activated charcoal and added with 1 mg l-1 2,4-D and 0.1 mg l-1 kinetin. The results showed that at all irradiation dosages, calli biomass increased significantly. The highest proliferation of calli biomass of 5.33 folds from the initial culture after 4 weeks was achieved at gamma irradiation of 25 Gy, whereas the lowest proliferation of calli biomass of 3.4 folds was achieved at control. The best development of embryogenic calli was obtained at 10 Gy that produced 100% somatic embryos, whereas the lowest somatic embryo formation at 0% was obtained at 0 and 25 Gy after one subculture. High response of somatic embryo induction to gamma irradiation at 10 Gy may increase production of somatic embryos. These results can be used in in vitro breeding of sago palm via mutagenesis to create new elite varieties.

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Perkembangan kalus embriogenik sagu (Metroxylon sagu Rottb.) pada tiga sistem kultur in vitro Development of embryogenic callus of sago (Metroxylon sagu Rottb.) on three systems of in vitro culture

E-Journal Menara Perkebunan ◽

10.22302/ppbbi.jur.mp.v76i1.88 ◽

2016 ◽

Vol 76 (1) ◽

Author(s):

Pauline D KASI ◽

. SUMARYONO

Keyword(s):

In Vitro Culture ◽

Liquid Medium ◽

Embryogenic Callus ◽

Somatic Embryos ◽

Temporary Immersion System ◽

Embryogenic Cells ◽

Temporary Immersion ◽

Friable Embryogenic Callus ◽

Metroxylon Sagu

Summary Embryogenic callus of sago (Metroxylon sagu Rottb.) has been grown on three systems of in vitro culture i.e. agar-solidified medium, liquid medium, and temporary immersion system (TIS) medium to observe and compare the development of embryogenic callus over one passage of six weeks. A-half gram of embryogenic callus was cultured on a modified MS medium containing 10 mg/L 2,4-D and 0.1 mg/L kinetin. For histological studies, embryogenic callus was fixed in FAA and embedded in paraplast wax. Serial sections were stained with safranin 1% and observed microscopically. By the end of culture period, the development of embryogenic callus in TIS medium was relatively better than those of the other two media. Fresh weight of callus in liquid medium and TIS increased by 6.5-fold, while on agar-solidified medium increased by 5.4-fold in six weeks. About 40% of callus in liquid medium and TIS and 20% of callus on agar solidified medium have changed into somatic embryos at globular stage. Histology structure of embryogenic callus of the three systems of in vitro culture shows different pattern. On agar-solidified medium, secondary callus and friable embryogenic callus that consist of meristematic cells were formed. In contrast, more embryogenic cells were formed in liquid medium and TIS to support maturation process to somatic embryos. Therefore, temporary immersion system and liquid medium are recommended for maturation of embryogenic callus, whereas agar-solidified medium is for proliferation of embryogenic callus of sago. Ringkasan Kalus embriogenik sagu (Metroxylon sagu Rottb.) telah ditumbuhkan pada tiga sistem kultur in vitro yaitu medium padat, medium cair, dan medium dengan sistem perendaman sesaat (SPS) untuk mempelajari dan mem-bandingkan perkembangan dari kalus embrio-genik selama periode enam minggu. Setengah gram kalus embriogenik dikulturkan pada medium MS modifikasi yang mengandung 2,4-D 10 mg/L dan kinetin 0,1 mg/L. Untuk studi histologi, kalus embriogenik difiksasi dengan FAA dan embedding menggunakan lilin paraplast. Irisan diwarnai dengan safranin 1% dan diamati menggunakan mikroskop. Pada akhir periode kultur, pertumbuhan kalus pada medium dengan SPS lebih baik dibandingkan dengan medium cair dan padat. Bobot basah kalus pada medium cair dan SPS meningkat 6,5 kali sedangkan pada medium padat meningkat 5,4 kali dalam waktu enam minggu. Sebanyak 40% kalus pada medium cair dan SPS serta 20% kalus pada medium padat berubah menjadi embrio somatik fase globuler. Struktur histologi kalus embriogenik pada ketiga jenis sistem kultur in vitro menunjukkan pola yang berbeda. Pada medium padat terjadi pembentukan kalus sekunder dan kalus embriogenik remah yang terdiri atas sel-sel meristematik. Sebaliknya pada medium cair dan SPS pembentukan sel embriogenik lebih banyak yang menunjang proses pendewasaan menjadi embrio somatik. Oleh karena itu, medium cair dan SPS direkomendasikan untuk pendewasaan kalus embriogenik, sedangkan medium padat untuk proliferasi kalus embriogenik sagu.

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Induksi dan Proliferasi Embriogenesis Somatik In Vitro pada Lima Genotipe Kedelai

Jurnal Agronomi Indonesia (Indonesian Journal of Agronomy) ◽

10.24831/jai.v44i3.12782 ◽

2017 ◽

Vol 44 (3) ◽

pp. 261

Author(s):

Adam Saepudin ◽

Nurul Khumaida ◽

Didy Sopandie ◽

Dan Sintho Wahyuning Ardie

Keyword(s):

Somatic Embryo ◽

Embryogenic Callus ◽

Somatic Embryos ◽

Induction Medium ◽

Embryo Induction ◽

Soybean Genotypes ◽

Induction Experiment ◽

Embryogenic Callus Induction ◽

Somatic Embryo Induction

ABSTRACT Somatic embryo induction medium was reported to be genotype dependent for soybean. This study was aimed to obtain the optimum medium for embryo somatic induction and proliferation, and to regenerate somatic embryo of five soybean genotypes. Five soybean genotypes (Tanggamus, Anjasmoro, Yellow Biloxi, CG-22-10, and SP-10-4) were used in this study. The research was divided into four steps: (1) embryogenic callus induction of five soybean genotypes, (2) embryogenic callus proliferation of five soybean genotypes, (3) optimation of embryo somatic induction on five soybean genotypes and (4) embryo somatic regeneration of five soybean genotypes. The induction experiment showed that based on number of embryogenic callus, the best somatic embryo-induction medium was 3% sucrose+ NAA 5 mg L-1+2,4-D 5 mg L-1+ Vitamin B5. Embryogenic callus number for each genotype tested was increased on proliferation media of 3% sukrosa + 2,4-D 5 mg L-1 + NAA 5 mg L-1+ Vit B5, and Yellow Biloxi gave the highest number of proliferated somatic embryos compared to other genotypes. Increasing number of globular somatic embryo of all genotypes was obtained from the optimation of somatic embryo induction media being used, and Tanggamus genotype gave the highest number of globular somatic embryo which followed by Yellow Biloxi genotype. Tanggamus and Yellow Biloxi genotypes were also successfully formed the four steps of somatic embryos (globular, heart, torpedo, and cotyledonary stages), but in regeneration medium of MS0 and media MS + sukrosa 10 g L-1 + GA3 2 mg L-1 + BAP 4 mg L-1 + Vit B5 only Tanggamus genotype was regenerated into plantlet. Keywords: 2,4-D, NAA, somatic embryos, induction, proliferation

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Perkembangan kalus embriogenik sagu (Metroxylon sagu Rottb.) pada tiga sistem kultur in vitro Development of embryogenic callus of sago (Metroxylon sagu Rottb.) on three systems of in vitro culture

E-Journal Menara Perkebunan ◽

10.22302/iribb.jur.mp.v76i1.88 ◽

2016 ◽

Vol 76 (1) ◽

Author(s):

Pauline D KASI ◽

. SUMARYONO

Keyword(s):

In Vitro Culture ◽

Liquid Medium ◽

Embryogenic Callus ◽

Somatic Embryos ◽

Temporary Immersion System ◽

Embryogenic Cells ◽

Temporary Immersion ◽

Friable Embryogenic Callus ◽

Metroxylon Sagu

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Shoot apical development during in vitro embryogenesisThis review is one of a selection of papers published on the Special Theme of Shoot Apical Meristems.

Canadian Journal of Botany ◽

10.1139/b06-070 ◽

2006 ◽

Vol 84 (11) ◽

pp. 1650-1659 ◽

Cited By ~ 8

Author(s):

Muhammad Tahir ◽

Claudio Stasolla

Keyword(s):

Shoot Apical Meristem ◽

Somatic Embryos ◽

Apical Meristem ◽

Flowering Plants ◽

Model Systems ◽

Marker Genes ◽

Zygotic Embryos ◽

Stage Of Development ◽

Meristem Development

Formation of the shoot apical meristem (SAM) has been extensively investigated in zygotic embryos of flowering plants, where it follows a prolonged and dynamic developmental pattern underlined by precise temporal and spatial changes in gene expression. Studies conducted on the plant model system Arabidopsis have revealed that SAM formation is controlled by a genetic network and involves the participation of several regulatory genes expressed at different stages of development. As a general rule apical meristem development in vivo occurs very early; at the globular stage of development in flowering plants and in club-stage embryos of conifers. Once formed, meristems of zygotic embryos are stable structures that become reactivated at the onset of germination. Shoot apical meristem formation during in vitro embryogenesis is demarked by structural events similar to those described for zygotic embryos, although differences can be observed during the late phases of development, where cellular differentiation and formation of intercellular spaces disrupt the architecture of SAMs produced in culture. These events, which denote the “unstable” nature of SAMs of somatic embryos, often result in poor conversion frequency and reduced plant regeneration. By using Picea glauca (Moench) Voss (white spruce) somatic embryos and microspore-derived embryos of Brassica napus L. (canola) as model systems, this review provides methods for improving SAM formation through manipulations of the culture medium which alter the cellular redox status. Meristem marker genes from Arabidopsis, such as WUSCHEL (which is required for the acquisition of stem fate identity), represent a valuable tool for estimating the quality of SAM produced by microspore-derived embryos of canola. In spruce, the identification of two novel meristem marker genes, HBK1 and PgAGO, will allow similar studies in conifers.

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Pengaruh Kinetin dan BAP terhadap Pertumbuhan dan Perkembangan Embrio Somatik Tanaman Sagu (Metroxylon sagu Rottb.)

Jurnal AgroBiogen ◽

10.21082/jbio.v6n2.2010.p101-106 ◽

2016 ◽

Vol 6 (2) ◽

pp. 101

Author(s):

Imron Riyadi

Keyword(s):

Plant Growth ◽

Somatic Embryo ◽

Somatic Embryos ◽

Growth And Development ◽

Culture Medium ◽

Sago Palm ◽

Murashige Skoog ◽

Expression Rate ◽

The Media ◽

Metroxylon Sagu

Effect of Kinetin and BAP to Growth and Development of Somatic Embryos of Sago Palm (Metroxylon sagu Rottb.). Imron Riyadi. Somatic embryos induction in sago palm (Metroxylon sagu Rottb.) have succesfully developed. Kinds and concentration of plant growth regulators (PGR’s) influence to growth and development of somatic embryos. The research was conducted to determine the optimal concentration of BAP and kinetin for proliferation, maturation and germination of sago palm somatic embryos. Cotyledonstage of somatic embryo derived from shoot tip cultures cultured on Modified Murashige-Skoog (MMS) with halfstrength macro-salts and added with 30 g/l sucrose, 2 g/l gelrite, 1 g/l activated charcoal. pH of media was adjusted at 5.6 before sterilized. The media were supplemented with 0.1-2.0 mg/l BAP and 0.1-2.0 mg/l kinetin in combination with 0.01 mg/l ABA each for supporting growth and development. The cultures was incubated at 26+1oC under a 12-h photoperiod with lighting providing an intensity 20 μmoles photons/m2/second for 11 weeks with replication 10 times. The results showed that the highest of somatic embryo proliferation was achieved in a culture medium with BAP at 0.5 mg/l + 0.01 mg/l ABA with an expression rate of 94%, the best maturation at 1.0 mg/l kinetin + 0.01 mg/l ABA with an expression rate of 93.5% and the most germination at 2.0 mg/l kinetin + 0.01 mg/l ABA with an expression rate of 100%. Transfer of these germinants to gelled media without PGR’s led to the development of normal plantlets.

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Embriogenesis Somatik Jeruk Keprok (Citrus reticulata L. cv Batu 55) Asal Hasil Perlakuan Kolkisin

Jurnal Hortikultura Indonesia ◽

10.29244/jhi.6.3.161-171 ◽

2015 ◽

Vol 6 (3) ◽

pp. 161 ◽

Cited By ~ 1

Author(s):

Agus Purwito ◽

Mohamad Prayogi ◽

Mia Kosmiatin ◽

Ali Husni

Keyword(s):

Somatic Embryo ◽

Embryogenic Callus ◽

Somatic Embryos ◽

Shoot Elongation ◽

Experimental Unit ◽

Culture Vessel ◽

Ms Medium ◽

Randomized Design ◽

Better Than

ABSTRACT The objective of this study was to obtain the best method of regeneration through somatic embryogenesis of citrus cv Batu 55 from callus resulted from in vitro polyploidization by colchicine. The experiment was conducted at the Laboratory of ICABIOGRAD, Bogor and Tissue Culture Laboratory, Department of Agronomy and Horticulture, Bogor Agricultural University from March 2014 until September 2014. This study consisted of proliferation of embryogenic callus, maturation, germination of somatic embryos, growth of shoots and roots. The research were comprised of four experiments, namely: 1). The effect of Phytagel concentration (2.5, 3.0, 3.5 and 4.0 g L-1) on proliferation of embryogenic callus, with 3 replications. Each experimental unit consisted of 3 clumps of callus, 2). The effect of ABA concentration (0, 0.1, 0.3 and 0.5 mg L-1) on somatic embryo maturation with 6 replications. Each experimental unit was one culture vessel containing five somatic embryos at globular phase, 3) The effect of vitamin composition (vitamin MS and vitamin MW) on germination of somatic embryo with 16 replications. Each experimental unit was one culture vessel containing four somatic embryos at cotyledonary phase, and 4) The effect 0.5 mg L-1 of plant growth regulators (NAA, IAA, IBA) and vitamin (MS and MW) on rooting and shoot elongation of germinated somatic embryos. Experiment was repeated five times. Each experimental unit was one culture vessel containing one plantlet as explant. All experiments were arranged as a completely randomized design. The result showed that the best concentration of Phytagel for callus proliferation was 2.5 g L-1. Maturation of somatic embryos was better when the somatic embryos were planted on medium supplemented with ABA 0.5 mg L-1. The MS medium supplemented with vitamin MS was better than supplemented with vitamin MW for the formation of plantlets, while roots and shoots elongation of the plantlet was better when explant was planted on the MS medium supplemented with vitamins MS and IBA 0.5 mg L-1. Key words: proliferation, maturation, germination, embryogenic callus, plantlet. ABSTRAK Penelitian ini bertujuan untuk mendapatkan metode embriogenesis somatik terbaik dari kalus Jeruk Keprok cv Batu 55 yang mendapatkan perlakuan poliploidisasi dengan kolkisin. Penelitian dilakukan di Laboratorium Balai Besar Litbang Bioteknologi & Sumber Daya Genetik Pertanian, dan Laboratorium Kultur Jaringan, Departemen Agronomi dan Hortikultura, Institut Pertanian Bogor mulai bulan Maret 2014 hingga September 2014. Penelitian ini terdiri atas proliferasi kalus embriogenik, pendewasaan, perkecambahan embrio somatik (ES), pertumbuhan tunas dan akar. Penelitian terdiri atas empat percobaan, yaitu: 1). Pengaruh konsentrasi Phytagel (2.5, 3.0, 3.5 dan 4.0 g L-1) terhadap proliferasi kalus embriogenik, dengan 3 ulangan, dimana setiap satuan percobaan terdiri atas 3 klum kalus, 2). Pengaruh konsentrasi ABA (0, 0.1, 0.3 dan 0.5 mg L-1) terhadap pendewasaan ES dengan 6 ulangan. Setiap satuan percobaan ialah satu botol kultur yang ditanam lima ES fase globular, 3) Pengaruh komposisi vitamin (vitamin MS dan vitamin MW) terhadap perkecambahan ES dengan 16 ulangan. Setiap satuan percobaan ialah satu botol kultur yang ditanami empat ES fase kotiledon, dan 4) Pengaruh 0.5 mg L-1 zat pengatur tumbuh (NAA, IAA, IBA) dan vitamin (MS dan MW) terhadap pertumbuhan tunas dan akar pada ES yang telah berkecambah. Setiap perlakuan diulang lima kali. Setiap satuan percobaan ialah satu botol kultur yang berisi satu planlet. Seluruh percobaan disusun menggunakan rancangan acak lengkap (RAL). Hasil menunjukkan bahwa konsentrasi terbaik Phytagel untuk proliferasi kalus adalah 2.5 g L-1. Pendewasan menjadi ES fase kotiledon akan lebih baik jika ES ditanam pada medium dengan ABA 0.5 mg L-1. Untuk pembentukan planlet, ES fase kotiledon akan lebih baik ditanam dalam medium MS yang ditambah vitamin MS dibanding yang ditanam pada medium MS ditambah vitamin MW. Medium untuk pertumbuhan tunas dan akar terbaik adalah medium MS yang ditambah dengan vitamin MS dan IBA 0.5 mg L-1. Kata kunci: proliferasi, pendewasaan, perkecambahan, kalus embriogenik, planlet.

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Effects of Plant Growth Regulators on In vitro Regeneration of Malaysian Indica rice (Oryza sativa L.) cv. MR219 by Shoot Apical Meristem

Asian Journal of Agricultural Research ◽

10.3923/ajar.2012.180.187 ◽

2012 ◽

Vol 6 (4) ◽

pp. 180-187 ◽

Cited By ~ 1

Author(s):

Lavanya Silvarajan ◽

Rosimah Nulit ◽

Faridah Qamaruz Za

Keyword(s):

Oryza Sativa ◽

Plant Growth ◽

Plant Growth Regulators ◽

Growth Regulators ◽

Shoot Apical Meristem ◽

Apical Meristem ◽

Oryza Sativa L ◽

Indica Rice ◽

In Vitro Regeneration

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EFFECT OF DIFFERENT SOURCES OF PLANT GROWTH REGULATOR ON THE INDUCTION AND DEVELOPMENT OF MANGOSTEEN SOMATIC EMBRYOS

Indonesian Journal of Agricultural Science ◽

10.21082/ijas.v17n1.2016.p9-16 ◽

2017 ◽

Vol 17 (1) ◽

pp. 9

Author(s):

Yosi Zendra Joni ◽

Riry Prihatini ◽

Darda Efendi ◽

Ika Roostika

Keyword(s):

Somatic Embryogenesis ◽

Plant Growth ◽

Embryogenic Callus ◽

Plant Growth Regulator ◽

Somatic Embryos ◽

Casein Hydrolysate ◽

Malt Extract ◽

Mass Propagation ◽

Embryogenic Callus Induction

Somatic embryogenesis is a technique for regenerating embryos derived from somatic cells of various plant species. This technique along with the utilization of plant growth regulator (PGR) might benefit for mass propagation and improvement of plant species through biotechnological tools. The study aimed to determine the effect of different plant growth regu-lators, namely 6-benzyladenine (BA) and thidiazuron (TDZ) on the embryogenic callus induction as well as casein hydrolysate and malt extract on the somatic embryo development of mangosteen. The explants used were in vitro young stems of mangosteen clone Leuwiliang. This study consisted of two experiments, namely induction of embryogenic callus and formation of somatic embryo. The first experiment was arranged as factorial in a completely randomized design with BA (0 and 0.7 mg l-1) as the first factor and TDZ (0, 0.1, 0.5 and 1.0 mg l-1) as the second factor. The second experiment consisted of four treatments, i.e. casein hydrolysate and malt extract at the rate of 500 and 1,000 mg l-1. The results showed that the best medium for embryogenic callus induction was MS supplemented with 0.1 mg l-1 TDZ, which resulted semifriable calli. Casein hydrolysate and malt extract could not induce the formation of somatic embryos. After two times subcultures on the same MS medium supplemented with 0.5 mg l-1 TDZ and 0.7 mg l-1 BA, a total of 33.8 somatic embryos per explant was induced. The successful somatic embryogenesis would support mangosteen breeding and in vitro mass propagation program.

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