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Fitri Rachmawati
Indonesia Ornamental Crop Research Institute (IOCRI)
Agriculture, Biological Sciences & Forestry

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Pengaruh Jenis Eksplan dan Asam Amino pada Inisiasi dan Proliferasi Kalus Embriogenik Phalaenopsis Var. ‘Raiza Agrihorti’ Fitri Rachmawati; Dedeh Siti Badriah; Budi Marwoto
Jurnal Hortikultura Vol 31, No 1 (2021): Juni 2021
Publisher : Indonesian Center for Horticulture Research and Development

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/jhort.v31n1.2021.p11-20

Abstract

(The Effect of Explant Types and Amino Acids on Embryogenic Callus Initiation and Proliferation of Phalaenopsis Var. ‘Raiza Agrihorti’)Penyiapan kalus embriogenik (KE) yang optimal memiliki peranan penting dalam menghasilkan benih bermutu Phalaenopsis skala komersial. Kendala utama yang dihadapi ialah inisiasi dan proliferasi KE yang masih rendah, serta akumulasi fenolik yang tinggi. Penelitian dilakukan di Laboratorium Kultur Jaringan Balithi dari Agustus 2019 hingga Juli 2020. Penelitian menggunakan Rancangan Acak Kelompok (RAK) pola split plot dan faktorial dengan lima ulangan. Percobaan-1: jenis eksplan (pucuk, pangkal, dan daun plantlet) sebagai petak utama dan perlakuan asam amino (tanpa asam amino, L-Proline, L-Glutamine, L-Cysteine, dan Casein-Hydrolisate) dengan konsentrasi 150 mg/l pada medium PC1 (1/2 MS + 1,0 mg/l TDZ + 0,5 mg/l BAP + 20 g/l sukrosa) sebagai anak petak. Percobaan-2: faktor-1 ialah jenis asam amino (L-Proline, L-Cysteine; L-Glutamine, dan Casein-Hydrolisate) dan faktor-2 ialah konsentrasi asam amino (0, 75, 150, 225, dan 300 mg/l). Hasil penelitian menunjukkan bahwa inisiasi KE Phalaenopsis var. ‘Raiza Agrihorti’ terbaik didapatkan dari pangkal plantlet dan 150 mg/l L-Glutamine dengan waktu inisiasi 18,3-24,0 hari, 80-100% pembentukan KE, dan ukuran KE 0,4-0,5 cm3. Proliferasi KE terbaik ditemukan pada L-Glutamine dengan konsentrasi 150 mg/l. Proliferasi KE mencapai 100% dengan penambahan berat segar sebesar 0,39 g, tingkat multiplikasi (MR) 4,55 kali dan pencokelatan 4,0%. Hasil penelitian ini berpotensi tinggi untuk diterapkan pada kultur starter Phalaenopsis hibrida lain.KeywordsPhalaenopsis hibrid; Asam amino; Inisiasi; Kalus embriogenik; ProliferasiAbstractSetup of the optimum Phalaenopsis embryogenic callus (EC) is an important role in producing qualified-seedlings of Phalaenopsis in commercial scale. The main constraints that are still being faced are the low rate of culture proliferation and high phenolic accumulations. The research was carried out at the Tissue Culture Laboratory-Indonesian Ornamental Plants Research Institite, from August 2019 through July 2020. The split plot and factorial designs were arranged using a Randomized Completely Block Design (RCBD) with five replications. Experiment-1: explants type (shoot tip, basal part, and leaf of plantlet) was used as main plot and amino acids (amino acids free, L-Proline, L-Glutamine, L-Cysteine, and Casein-Hydrolisate) with 150 mg/l concentration on medium PC1 (1/2 MS + 1,0 mg/l TDZ + 0,5 mg/l BAP + 20 g/l sukrosa) as subplot. Experiment-2: the first factor was amino acids type (L-Proline, L-Cysteine; L-Glutamine, and Casein-Hydrolisate) and the second factor was amino acids concentration (0, 75, 150, 225, and 300 mg/l). Results of the studies revealed that the best EC initiation of Phalaenopsis var. ‘Raiza Agrihorti’ was produced by basal part of plantlet and PC1 medium containing 150 mg/l L-Glutamine with EC Initiation time was 18.3-24.0 days, 80-100% of EC formation and size of 0.4-0.5 cm3. The best proliferation of EC was found in L-Glutamine with 150 mg/l concentration. EC proliferation reached 100% with 4.55 EC multiplication rate, 0.39 g EC fresh weight added, and EC browning as low as 4.0%. The established method is high possibly applied for other Phalaenopsis hybrids.
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) Fitri Rachmawati; Dewi Permanik; Ronald Bunga Mayang; Budi Winarto
Jurnal Hortikultura Vol 29, No 2 (2019): Desember 2019
Publisher : Indonesian Center for Horticulture Research and Development

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/jhort.v29n2.2019.p137-146

Abstract

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.
Co-Authors Budi Marwoto Budi Winarto Dedeh Siti Badriah Dewi Permanik Ronald Bunga Mayang