Shoot Tips Derived-Somatic Embryogenesis in Mass Propagation of 
Dendrobium Indonesia Raya  and #8216;Ina and #8217;

Rachmawati, Fitri; Purwito, Agus; Wiendy, Ni Made Armini; Mattjik, Nurhayati Ansori; Winarto, Budi

doi:10.9755/ejfa.2015-05-212

Cited by 7 publications

(11 citation statements)

References 18 publications

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“…Percobaan disusun menggunakan rancangan acak kelompok (RAK) faktorial, dua faktor dengan lima ulangan. Faktor pertama adalah tiga media inisiasi, yaitu MI-1 (1,5 mg/l TDZ + 0,5 mg/l BAP) sebagai kontrol (Rachmawati et al 2015& Rachmawati et al 2016), MI-2 (2,5 mg/l metathopolin (mT) + 0,05 mg/l BAP), dan MI-3 (5,0 mg/l mT + 0,05 mg/l BAP). Semua perlakuan menggunakan media dasar setengah kekuatan Murashige and Skoog (½ MS) dengan penambahan 20 g/l sukrosa dan 2 g/l gelrite.…”

Section: Inisiasi Kalus Embriogenik D 'Balithi Cf22-58'unclassified

“…Ketersediaan teknologi perbanyakan yang efektif dan efisien merupakan salah satu komponen penting untuk mendukung komersialisasi produk pemuliaan, khususnya terkait dengan penyediaan benih berkualitas. Protokol perbanyakan masal secara in vitro berbasis embrio somatik telah berhasil dikembangkan dan dilaporkan oleh yang berhasil mengembangkan protokol tersebut untuk Dendrobium 'Zahra FR-62' dan D. 'Gradita 31' ), D. 'Gradita 10' (Rachmawati et al 2014), dan D. Indonesia Raya 'Ina' (Rachmawati et al 2015), D. Sonia-28 (Julkiflee, Uddain & Subramaniam 2014), D. Sonia 'Ersakul' (Juntada et al 2015), dan D. aqueum (Parthibhan et al 2018). Protokol tersebut umumnya dimulai dari inisiasi embriogenesis, baik langsung maupun tidak langsung melalui kultur eksplan terpilih pada medium terseleksi, dilanjutkan dengan tahap perbanyakan/proliferasi kalus embriogenik dan atau embrio, pembentukan dan perkembangan embrio, perkecambahan embrio, penyiapan plantlet hingga aklimatisasinya.…”

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Protokol Perbanyakan Masal Dendrobium ‘Balithi CF22-58’ secara In Vitro Melalui Embriogenesis Somatik Tidak Langsung (In Vitro Propagation Protocol of Dendrobium ‘Balithi CF22-58’ via Indirect Somatic Embryogenesis)

Rachmawati¹,

Permanik²,

Mayang³

et al. 2020

J.Hort

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Protokol perbanyakan klonal yang efektif dan efisien sangat diperlukan untuk produksi benih berkualitas pada komersialisasi produk unggulan hasil pemuliaan. Penelitian bertujuan untuk mendapatkan protokol perbanyakan klonal Dendrobium ‘Balithi CF22-58’ melalui embriogenesis tidak langsung. Percobaan dilakukan di Laboratorium Kultur Jaringan Balai Penelitian Tanaman Hias dari bulan Januari hingga Desember 2017. Penelitian ini menekankan pada penggunaan jenis eksplan, media, dan sistem kultur. Jenis eksplan yang diuji adalah tunas pucuk, tunas lateral, dan pangkal plantlet dengan tiga media inisiasi [½ Murashige and Skoog (MS) dikombinasikan dengan 1,5 mg/l thidiazuron (TDZ) dan 0,5 mg/l 6-benzylaminopurine (BAP) (MI-1), 2,5 mg/l metathopolin (mT) dan 0,05 mg/l BAP (MI-2), dan 5 mg/l mT dan 0,05 mg/l BAP (MI-3)]; empat media proliferasi, yaitu ½ MS dengan kombinasi: MP-1 (0,75 mg/l TDZ + 0,25 mg/l BAP), MP-2 (1,5 mg/l TDZ + 0,5 mg/l BAP), MP-3 (2,5 mg/l mT + 0,05 mg/l BAP), dan MP-4 (5,0 mg/l+ 0,05 mg/l BAP); dua sistem kultur (padat dan cair); dan tiga media regenerasi MPP-1 (½ MS dengan vitamin penuh (1/2 MS-FV) + 2% charcoal); MPP-2 (½ MS-FV); dan MPP-3 (2 g/l Rosasol 18:18:18 TE). Percobaan disusun menggunakan rancangan acak kelompok faktorial dengan lima ulangan. Hasil penelitian menunjukkan bahwa inisiasi kalus embriogenik (KE) tertinggi, yaitu 38,3% dengan waktu inisiasi 16,8 hari dihasilkan dari eksplan pangkal plantlet pada medium MI-1. Medium MP-2 dan sistem kultur cair mampu mempertahankan proliferasi KE sampai 83,1% dengan rasio penggandaan 3,23 kali. Perkecambahan embrio terbaik sampai 86,9% embrio berkecambah dengan 18,2 kecambah per rumpun dalam waktu 21,3 hari, ditunjukkan pada medium MPP-1, sedangkan pembesaran plantlet terbaik mencapai tinggi plantlet sampai 5 cm, jumlah daun hingga 4,9 helai, dan jumlah akar 2,8, dengan 2,6 cm panjang akar dan 0,27 g bobot basah plantlet, diperoleh pada medium MPP-3. Perbanyakan anggrek dengan protokol ini diperkirakan dapat menghasilkan sekitar 3.000–4.000 plantlet/eksplan/tahun. Protokol hasil penelitian ini sangat potensial diaplikasikan pada perbanyakan klonal Dendrobium melalui kultur jaringan. KeywordsDendrobium; Embriogenesis somatik; Perbanyakan masal; Proliferasi; Sistem kultur AbstractThe effective and efficient clonal propagation protocol is significantly needed for producing qualified seedling for commercialization of superior breeding products. The objective of the study was to establish clonal propagation protocol for Dendrobium ‘Balithi CF22-58’ via indirect somatic embryogenesis. The study was conducted at the Tissue Culture Laboratory in Indonesian Ornamental Crops Research Institute from January to December 2017. The study emphasized to utilize explant source, culture media, and culture system. Explant types were shoot tip, lateral shoot, and basal part of plantlets; three initiation media [half strength Murashige and Skoog (MS) medium containing 1.5 mg/l thidiazuron (TDZ) and 0.5 mg/l 6-benzylaminopurine (BAP) (MI-1), 2.5 mg/l metathopolin (mT) and 0.05 mg/l BAP (MI-2), and 5 mg/l mT and 0.05 mg/l BAP (MI-3)]; four proliferation media (half strength MS medium supplemented with: MP-1 (0.75 mg/l TDZ and 0.25 mg/l BAP), MP-2 (1.5 mg/l TDZ and 0.5 mg/l BAP), MP-3 (2.5 mg/l mT and 0,05 mg/l BAP), and MP-4 (5.0 mg/l and 0.05 mg/l BAP); two culture system were solid and liquid; and three regeneration media viz, MPP-1 (half strength MS medium with full vitamin and 2% activated charcoal); MPP-2 (MR-1 activated charcoal free), and MPP-3 (2 g/l Rosasol 18:18:18 TE). These experiments were arranged using a factorial randomized complete block design with five replications. Results of the study revealed that the highest initiation rate of embryogenic callus (EC) was up to 38.3% in 16.8 days after culture. The EC was regenerated from a basal part on MI-1 medium, MP-2 medium and liquid culture system were able to maintain proliferation of embryogenic callus up to 83.1% with 3.23 multiplication rate. The best embryo germination up to 86.9% with 18.2 germinated embryos per clump within 21.3 days was determined on MPP-1 medium. While the best plantlet performances with 5 cm height of plantlets, 4.9 number of leaves, 2.8 number of roots, 2.6 cm root length, and 0.27 g plantlet fresh weight was obtained MPP-3 medium. With this propagation protocol, 3,000 - 4,000 plantlets/explant/year can be produced. Results of the study have high potential to be applied for in vitro propagation of Dendrobiums.

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Section: Inisiasi Kalus Embriogenik D 'Balithi Cf22-58'unclassified

unclassified

Protokol Perbanyakan Masal Dendrobium ‘Balithi CF22-58’ secara In Vitro Melalui Embriogenesis Somatik Tidak Langsung (In Vitro Propagation Protocol of Dendrobium ‘Balithi CF22-58’ via Indirect Somatic Embryogenesis)

Rachmawati¹,

Permanik²,

Mayang³

et al. 2020

J.Hort

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“…Proliferasi kalus selanjutnya dilakukan dalam erlenmeyer 100 ml berisi 25 ml media cair PM-12 (medium ½ MS + 0.5 mg L -1 TDZ + 0.10 mg L -1 BA + 150 mL L -1 AK) dengan kepadatan kalus 3 g dan kultur digojok di atas orbital shaker dengan kecepatan ± 100 rpm, disubkultur 3-4 kali dengan interval 1 bulan (Gambar 1B). Semua tahapan kultur diinkubasi pada kondisi terang 12 jam di bawah lampu fluorescent dengan intensitas cahaya 13 µmol m -2 s -1 pada 23.5 ± 1.1 °C (Rachmawati et al, 2015). Kalus yang tumbuh dan berproliferasi dengan baik digunakan sebagai kultur starter untuk optimasi perbanyakan masal KE menggunakan bioreaktor.…”

Section: Bahan Dan Metodeunclassified

“…Kalus yang dipanen dari bioreaktor dibilas dengan aquadest steril dan ditempatkan di atas kertas tissue steril dalam cawan petri, kemudian disimpan selama 3-7 hari pada suhu 18 ± 2 °C (Rachmawati et al, 2015). Percobaan menggunakan Rancangan Acak Lengkap (RAL) dengan delapan perlakuan media perkecambahan (MK), yaitu dua media dasar (½ MS dan 2 g L -1 pupuk majemuk NPK dengan perbandingan 32:10:10) dikombinasikan dengan empat perlakuan (tanpa zat pengatur tumbuh (zpt), 0.05 mg L -1 BA, 150 mL L -1 AK, dan kombinasi keduanya) dengan penambahan 20 g L -1 gula dan 7 g L -1 agar.…”

Section: Diferensiasi Dan Perkecambahan Embrio Somatik (Es) D Indoneunclassified

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Perbanyakan In Vitro Dendrobium Indonesia Raya ‘Ina’ melalui Embriogenesis Somatik Berbasis Sistem Bioreaktor

Rachmawati¹,

Wiendi²,

Mattjik³

et al. 2017

J Agron Indonesia

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An effective and efficient in vitro propagation system has important roles in preparing and producing high qualityseedlings of Dendrobium for commercial scale. The objective of this research was to establish an effective and efficient embryogenic callus (EC) proliferation method using bioreactor system and regeneration EC into plantlet for producing high quality seedlings of Dendrobium Indonesia Raya 'Ina'. Differences in callus densities (5, 10, 15, and 20

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Somatic embryogenesis from stem thin cell layers of Dendrobium aqueum

Parthibhan

Rao

Silva³

et al. 2018

Biologia plant.

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An efficient in vitro regeneration protocol through somatic embryogenesis was established from stem transverse thin cell layers (tTCLs) of Dendrobium aqueum Lindley, an imperiled orchid. This study outlines the induction and successive maturation stages of D. aqueum somatic embryos (SEs). The tTCLs (~ 0.5 mm thick) cultured on halfstrength Murashige and Skoog (MS) medium containing cytokinins and auxins, either individually or in combination, produced embryogenic callus (EC). Treatment with 0.5 mg dm -3 zeatin induced EC in 41.42 % of tTCLs. As many as 42.66 globular SEs per tTCL were formed in the presence of 1.5 mg dm -3 N 6 -(2-isopentyl) adenine (2iP) but only on 10.33 % of explants. The combined treatment of 2iP (1.5 mg dm -3 ) and 0.5 mg dm -3 6-benzyladenine resulted in 34 globular SEs on 14.7 % of tTCLs whereas the combination of 2iP and 1.0 mg dm -3 indole-3-butyric acid (IBA) induced 7.4 globular SEs on 52.33 % of tTCLs. Supplementation of activated charcoal, amino acids, and antioxidants alleviated browning at all the concentrations tested, but the EC response declined. The addition of 0.5 mg dm -3 polyvinylpyrrolidone to 1.5 mg dm -3 2iP and 1.0 mg dm -3 IBA produced 24 SEs on 19.89 % of tTCLs suggesting that the EC and SEs can be effectively induced by individual cytokinins whereas the synergistic treatments with other compounds can only enhance the induction of EC. Histological observations of EC showed the formation of globular SEs from sub-epidermal regions. Successive developmental stages of globular SEs and the intermediate stage of protocorm like bodies until the formation of plantlets were observed. The plantlets obtained through SEs showed no morphological variations, and inter simple sequence repeat profiles also confirmed the genetic fidelity of in vitroderived progeny with high monomorphism (97.78 %). In conclusion, the use of stem tTCLs is an effective method to produce SEs through indirect somatic embryogenesis in D. aqueum.

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Shoot Tips Derived-Somatic Embryogenesis in Mass Propagation of Dendrobium Indonesia Raya and #8216;Ina and #8217;

Cited by 7 publications

References 18 publications

Protokol Perbanyakan Masal Dendrobium ‘Balithi CF22-58’ secara In Vitro Melalui Embriogenesis Somatik Tidak Langsung (In Vitro Propagation Protocol of Dendrobium ‘Balithi CF22-58’ via Indirect Somatic Embryogenesis)

Protokol Perbanyakan Masal Dendrobium ‘Balithi CF22-58’ secara In Vitro Melalui Embriogenesis Somatik Tidak Langsung (In Vitro Propagation Protocol of Dendrobium ‘Balithi CF22-58’ via Indirect Somatic Embryogenesis)

Perbanyakan In Vitro Dendrobium Indonesia Raya ‘Ina’ melalui Embriogenesis Somatik Berbasis Sistem Bioreaktor

Somatic embryogenesis from stem thin cell layers of Dendrobium aqueum

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