Anthurium anther culture was successfully established using half-anthers as explants. Explants were cultured on Winarto-Teixeira basal medium (WT-1) containing 0.01 mg/l a-naphthalene acetic acid (NAA), 0.5 mg/l thidiazuron (TDZ), and 1.0 mg/l 6-benzylaminopurine (BAP), or on New Winarto-Teixeira basal medium (NWT-3) supplemented with 0.02 mg/l NAA, 1.5 mg/l TDZ, and 0.75 mg/l BAP for callus initiation. Regenerated calli produced multiple shoots on WT-1, which were then rooted in NWT-3 supplemented with 1% activated charcoal. Plantlets were acclimatized ex vitro using a mixture of burned rice husk, rice husk, and bamboo peat (1:1:1, v/v/v) as the potting medium. There was considerable morphological and cytological diversity of regenerants derived from anther culture, which are described in detail in this study. The callus cluster color ranged from green to light green and had a high regeneration capacity (7.3 and 4.8 shoots/callus cluster), light reddish-yellow callus showed moderate regeneration (2.6 shoots/callus cluster), while reddish-yellow callus had the lowest regeneration capacity (1.5 shoots/callus cluster). Morphological variations clearly observed in regenerants derived from this technique included alterations in plant size, peduncle length, spathe position compared to leaves, the type and number of buds, spathe and spadix color, and spadix length. There were also cytological variations in both in vitro and ex vitro regenerants of anther culture with 23-29% haploids, 5-10% aneuploids, 56-69% diploids, and 3-4% triploids. The results strengthen other studies in which the development of anther cultures, especially via callus formation, resulted in morphological and cytological alterations. These variations have been discussed to great length in this paper.

A successful protocol for high frequency callus induction and plant regeneration from Anthurium andreanum Linden ex André cv. Tropical half-anthers is described. Different variables using Winarto and Teixeira and Murashige and Skoog basal media supplemented with several plant growth regulators [2,4-dichlorophenoxy acetic acid (0.1-1.0 mg/l), a-naphthalene acetic acid (0.01-0.2 mg/l), thidiazuron (0.5-2.0 mg/l), 6-benzylaminopurine (0.5-1.0 mg/l), and kinetin (0.5-1.0 mg/l)] were tested for their ability to induce high frequency callusing in half-anthers, indirect regeneration and rooting of shoots. Basal medium, as well as the combination and concentration of hormones applied, had a significant effect on callus formation, shoot regeneration and adventitious root formation. Winarto and Teixeira-1, an original basal medium containing 0.01 mg/l a-naphthalene acetic acid, 0.5 mg/l thidiazuron and 1.0 mg/l 6-benzylaminopurine was suitable for callus formation while an improved basal medium i.e., New Winarto-Teixeira-3 supplemented with 0.25 mg/l 2,4-dichlorophenoxy acetic acid, 0.02 mg/l a-naphthalene acetic acid, 1.5 mg/l thidiazuron and 0.75 mg/l 6-benzylaminopurine enhanced callus formation. High shoot regeneration and multiplication was also possible on New Winarto-Teixeira-3. Shoots formed a strong adventitious root system on New Winarto-Teixeira-3 containing 0.2 mg/l a-naphthalene acetic acid and 1.0 mg/l kinetin. Plantlets that varied in size and performance were successfully acclimatized and adapted to ex vitro conditions. Cytological analysis of 180 acclimatized-plantlets ex vitro revealed that 34 were haploid (n = 14-18), 15 aneuploid (n = 20-26), 126 diploid (n = 28-34) and 5 triploid (n = 45-57). The potential use of this protocol for developing half-anther culture of other Anthurium species or cultivars is discussed.

An effective and efficient in vitro propagation system has important roles in preparing and producing high quality-seedlings 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 g callus in 250 mL medium), aeration levels (2.5, 5.0, and 10.0 O 2 volume per medium volume per minute; vvm), and regeneration media half-strength MS and 2 g L-1 NPK (32:10:10) combinated by 0.00, 0.05 mg L-1 BA, 150 mL L-1 coconut water and their combinations were tested in this experiment. The experiments were arranged using randomized completely block design (RCBD) with three replications for EC proliferation and randomized completely desaign (RBD) for EC regeneration. The results showed that combination of aeration at 2.5 vvm and 10 g of EC was the most suitable aeration level and callus density for proliferation of EC in the 500 ml airlift bioreactor with 6.85 multiplication rate, 92.5% EC formation, and malformed callus morphology as low as 6.1%. The highest somatic embryos (SEs) formation was 87.7% with 44.5 SEs per clump and 92.1% SEs germination with 41.0 germinated-SEs per clump, 85.1% normal germinated-SEs, and whereas the best performance of plantlet was obtained from 1/2 MS + 0.05 mg L-1 BA semi solid medium. Plantlets were successfully acclimatized using Cycas rumphii medium with high survival rate (91.6%). ABSTRAK Sistem perbanyakan in vitro yang efektif dan efisien memiliki peranan penting dalam mempersiapkan dan memproduksi benih Dendrobium bermutu untuk skala komersial. Penelitian bertujuan mendapatkan metode proliferasi kalus embriogenik (KE) menggunakan bioreaktor dan regenerasi KE menjadi plantlet yang efektif dan efisien untuk perbanyakan masal benih D. Indonesia Raya 'Ina' bermutu. Perbedaan kepadatan kalus (5, 10, 15, dan 20 g kalus dalam 250 mL media), tingkat aerasi (2.5, 5.0, dan 10.0 volume O 2 per volume media per menit; vvm), dan media regenerasi (media dasar ½ MS dan 2 g L-1 NPK (32:10:10) yang dikombinasikan 0.00, 0.05 mg L-1 BA, 150 mL L-1 air kelapa, dan kombinasi keduanya) diuji dalam penelitian ini. Percobaan menggunakan Rancangan Acak Kelompok Lengkap (RAKL) pola faktorial dengan tiga ulangan untuk proliferasi KE dan Rancangan Acak Lengkap (RAL) dengan tiga ulangan untuk regenerasi KE menjadi plantlet. Hasil penelitian menunjukkan bahwa kombinasi tingkat aerasi 2.5 vvm dengan kepadatan kalus 10 g paling sesuai untuk proliferasi KE dalam airlift bioreactor 500 ml dengan 6.85 tingkat multiplikasi, 92.5% pembentukan KE, dan 6.1% malformasi morfologi kalus. Diferensiasi dan perkecambahan embrio somatik (ES) tertinggi (87.7%) dengan 44.5 ES per gerombol, 92.1% ES berkecambah dengan 41.0 kecambah per gerombol, 85.1% kecambah normal, dan menghasilkan pertumbuhan plantlet terbaik ditemukan pada media semi-solid 1/2 MS + 0.05 mg ...

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Ketersediaan protokol perbanyakan massa anggrek Dendrobium secara <em>in vitro</em> memiliki peranan penting dalam mendukung pengembangan industri benih di dalam negeri. Penelitian ini bertujuan mendapatkan teknologi perbanyakan massa Dendrobium Gradita 10 melalui embriogenesis somatik. Penelitian dilaksanakan di Laboratorium Kultur Jaringan dan Rumah Kaca Anggrek Balai Penelitian Tanaman Hias Segunung, Pacet, Cianjur, mulai bulan Maret sampai dengan Desember 2012. Penelitian<br />disusun menggunakan rancangan acak kelompok dengan lima ulangan. Jenis eksplan, media, periode subkultur, dan kepadatan kalus diujicobakan dalam penelitian ini. Hasil penelitian menunjukkan bahwa daun planlet dan media ½ Murashige &amp; Skoog (MS) + 1 mg/l Thidiazuron (TDZ) + 0,5 mg/l N6-benzyladenine (BA) merupakan jenis eksplan dan media terbaik untuk induksi kalus embriogenik hingga 80% dengan waktu pembentukan kalus tercepat (26,3 hari setelah kultur). Proliferasi kalus embriogenik terbaik terdapat pada media ½ MS + 0,3 mg/l TDZ + 0,1 mg/l α-naphthalene acetic acid (NAA) dengan kepadatan kalus 2–3 g kalus/25 ml medium. Pertumbuhan kalus embriogenik teroptimal terdapat pada periode subkultur yang ke-2. Konversi kalus embriogenik menjadi embrio somatik mencapai 79% pada subkultur ke-3 ditemukan pada media ½ MS + 0,05 mg/l BA. Perkecambahan embrio maksimal dengan 21,7 planlet per gerombol embrio ditemukan pada media ½ MS + 0,05 mg/l BA. Keberhasilan pengembangan teknologi perbanyakan massa anggrek Dendrobium Gradita 10 secara in vitro melalui embriogenesis somatik diharapkan memiliki dampak besar terhadap pengembangan teknologi perbanyakan massa benih untuk jenis Dendrobium yang lain.

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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<p>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. </p><p><strong>Keywords</strong></p><p><em>Dendrobium</em>; Embriogenesis somatik; Perbanyakan masal; Proliferasi;  Sistem kultur  </p><p><strong>Abstract</strong></p><p>The 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.</p>