scholarly journals 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)

2020 ◽  
Vol 29 (2) ◽  
pp. 137
Author(s):  
Fitri Rachmawati ◽  
Dewi Permanik ◽  
Ronald Bunga Mayang ◽  
Budi Winarto
Keyword(s):  
Somatic Embryogenesis ◽  
Activated Charcoal ◽  
Embryogenic Callus ◽  
Culture System ◽  
In Vitro Propagation ◽  
Clonal Propagation ◽  
Ms Medium ◽  
Indirect Somatic Embryogenesis ◽  
Half Strength

<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>

scholarly journals Clonal Propagation of Phalaenopsis amabilis (L.) BL. cv. 'Golden Horizon' Through In vitro Culture of Leaf Segments

1970 ◽  
Vol 46 (2) ◽  
pp. 163-168 ◽  
Author(s):  
P Sinha ◽  
MAA Jahan
Keyword(s):  
In Vitro Culture ◽  
Natural Environment ◽  
Activated Charcoal ◽  
Clonal Propagation ◽  
Ms Medium ◽  
Huge Amount ◽  
Leafy Shoots ◽  
Half Strength ◽  
Leaf Segments

A protocol was established for mass clonal propagation of Phalaenopsis amabilis cv. 'Golden horizon' through in vitro culture of young leaf segments from mature plant. Explants were cultured on half strength Murashige and Skoog (1/2 MS) medium supplemented with N6-benzyladenine (2.0 mg l-1), a-naphthaleneaceetic acid (0.5 mg l-1), 2% (w/v) sucrose, 10% (v/v) coconut water, 2 g l-1 peptone and 1 g l-1 activated charcoal. Each section of explant produced 15 protocorm-like bodies (PLBs) after 12 weeks of culture. When phytohormone was omitted from the medium and 150 mg l-1 L-glutamone was added PLBs were found to be enlarged with leafy shoots and new PLBs were induced from the base of the old ones. Leafy shoots rooted on half strength MS medium supplemented with 2 g l-1 peptone, 2% (w/v) sucrose, 10% (v/v) CW and 1 g l-1 activated charcoal, where 100% explants were developed into plantlets with roots within 8 weeks. The addition of 2.5 g l-1 banana pulp powder enhanced the number and length of roots. Within the first 32 weeks after initiation of culture about 1500 plantlets as well as a huge amount of PLBs were achieved from a single explant section. The plantlets were acclimatized in natural environment. Key words: Phalaenopsis orchid; Leaf segments; Protocorm-like bodies; Micropropagation DOI: http://dx.doi.org/10.3329/bjsir.v46i2.8182 Bangladesh J. Sci. Ind. Res. 46(2), 163-168, 2011

scholarly journals In Vitro Propagation of Croton Scabiosus Bedd. (Euphorbiaceae), an Endemic and Vulnerable Tree Species

2013 ◽  
Vol 3 (3) ◽  
pp. 229-241
Author(s):  
B. Ravi Prasad Rao ◽  
S. Salamma
Keyword(s):  
Tree Species ◽  
In Vitro Propagation ◽  
Andhra Pradesh ◽  
Natural Habitat ◽  
Eastern Ghats ◽  
Ms Medium ◽  
Species Population ◽  
Half Strength ◽  
Site Conservation

Croton scabiosus Bedd. (Euphorbiaceae), an endemic tree species of Southern Eastern Ghats of Andhra Pradesh, India, categorised as ‘Vulnerable’ is attempted for in vitro propagation. Seed pathological problems, poor germination, recurrent fires and tree fellings in the native habitat are the major threats of the species. With an objective to standardise a suitable multiplication protocol to augment the species population in the natural habitat, the present study focus on multiplication of the species through micropropagation. Of the various growth regulators employed in the experimentation, maximum mean number of shoots i.e. 5.37±0.12 was found on MS medium fortified with 0.5mg/l BAP and 2.5mg/l IAA. Maximum mean length of shoots 5.27±0.14 was found on 0.5mg/l BAP+1mg/l IAA. In vitro rooting was found on half strength MS medium fortified with 2.5mg/l IBA (8.92±0.19 mean number of roots with 5.04±0.05 mean length of roots with 90% response). Only 10% of survival rate was observed and the species warrants effective on site conservation methods.

scholarly journals Pear in Vitro Propagation Using a Double-phase Culture System

HortScience ◽  
1991 ◽  
Vol 26 (1) ◽  
pp. 62-64 ◽  
Author(s):  
R. Rodriguez ◽  
C. Díaz-Sala ◽  
L. Cuozzo ◽  
G. Ancora
Keyword(s):  
Culture System ◽  
In Vitro Propagation ◽  
Callus Formation ◽  
Plantlet Regeneration ◽  
Ms Medium ◽  
Inhibition Of Proliferation ◽  
Double Phase ◽  
Phase Culture ◽  
Modified Ms Medium

Proliferation of Pyrus communis L. cv. Abate Fetel, Precoce Morettini, and Guyot was accomplished with a yield of 10 to 15 new shoots per explant. The in vitro procedure is based on the use of 6.7 μm BAP as an overlay on a modified MS medium. Rooting without callus formation was achieved by immersing the basal end in 5 μm IBA solution for 1 min. The possible inhibition of proliferation and plantlet regeneration by GA3 and IBA is discussed. Chemical names used: 6-benzylaminopurine (BAP); indole-3-butyric acid (IBA); gibberellic acid (GA3).

scholarly journals Effect of Different Concentrations of Plant Growth Regulators on in Vitro Propagation of Curcuma roscoeana Wall.

HortScience ◽  
1997 ◽  
Vol 32 (3) ◽  
pp. 460D-460
Author(s):  
Chamchuree Sotthikul ◽  
Pimchai Apavatjrut
Keyword(s):  
Plant Growth ◽  
In Vitro Propagation ◽  
Clonal Propagation ◽  
Stem Explants ◽  
Ms Medium ◽  
Chiang Mai ◽  
Perennial Plant ◽  
The Media ◽  
Low Rate

Curcuma roscoeana Wall. is a tuberous perennial plant with tuberous rhizomes. It is an endangered species. In nature, it has a very low rate of multiplication. Propagation of C. roscoeana in vitro was done by culturing 0.5 × 1.0-mm shoot tips from young buds onto modified Murashige and Skoog (MS)+ 0.25 mg/L kinetin. Stem explants 10.0 mm in size, measured from the base of the plantlets longitudinally cut in half, were used in the experiments. The first experiment was done by varying the concentration of both kinetin and NAA, in MS liquid medium, at 0–8.0 mg/L and 0–0.05 mg/L, respectively. There were no significant differences of kinetin and NAA concentrations on the number of plantlets obtained. The 0.5-mg/L kinetin treatment gave the highest yield in number of new plantlets (3.1 plantlets/cultured explant). In the second experiment, various concentrations of BAP from 0 to 8.0 mg/l were tested. 2.8–3.7 plantlets were formed in the media with 0.05–2.0 mg/L of BAP. The most-suitable concentration of BAP was at 1.0 mg/L, providing 3.7 plantlets/cultured explants. Kinetin or BAP alone could be used in MS medium for rapid clonal propagation of C. roscoeana. The rooted plantlets could be successfully transferred into growing pots. Acknowledgement: The studies were supported in part by The King's Initiative Centre for Fruit and Flower propagation and Development, Ban Rai, Chiang Mai.

scholarly journals n vitro micro propagation of Spilanthes acmella (L.) Murr

2014 ◽  
Vol 8 (1) ◽  
pp. 55-60
Author(s):  
Bushra M. Jaber Alwash ◽  
Ansaam Z. Jassim
Keyword(s):  
In Vitro Propagation ◽  
Indole Acetic Acid ◽  
Ms Medium ◽  
Indole Butyric Acid ◽  
Micro Propagation ◽  
Spilanthes Acmella ◽  
Half Strength ◽  
The Mean ◽  
Initiation Stage

This study was aimed to In vitro propagation of Spilanthes acmella L. Murr. It is a medicinal plant not cultivated in Iraq. Seeds were sterilized and cultured on MS medium. Indole acetic acid IAA, Benzyladenin BA growth regulators’ were used at the initiation stage. The combination between IAA and BA was used in multiplication stage. Indole butyric acid IBA was used for rooting the shoots. Results showed that 1.5% sodium hypochlorite for 15 min was very effective for disinfecting and survival. A node exhibited relatively highest response as compared with apical meristems and leaflets culture. Supplying the culture medium with 1mg/l. BA was effective for lateral shoot induction. The mean number of shoots obtained from nodes were 7.43 with a mean length 0.9 cm. Adding BA at 0.5, 1.0 or 1.5 and IAA at 0.1 mg/l. to the growth medium was effective for multiplication. Mean number of the developed shoots were 12.00, 10, 84, 10.00 respectively. Adding 0.1, 0.5, 1.0 mg/l IBA to the half strength MS medium was very effective in root formation which produced 45.0, 42.5, 40.0 roots respectively with mean length of 3.25, 3.80, 3.80 cm respectively. Results of acclimatization stage showed that addition of 1:1 Patmos and loamy soil gave the highest rate of survival 100% after 4 weeks of acclimatization. This study showed the ability of in vitro propagation of Spilanthes acmella (L.) Murr

scholarly journals Clonal Propagation of Rhynchostylis retusa (Lin.) Blume through in vitro Culture and their Establishment in the Nursery

2012 ◽  
Vol 22 (1) ◽  
pp. 1-11
Author(s):  
Pinaki Sinha ◽  
Miskat Ara Akhter Jahan
Keyword(s):  
Plant Tissue ◽  
Activated Charcoal ◽  
High Frequency ◽  
Nutrient Medium ◽  
Clonal Propagation ◽  
Nodal Segments ◽  
Nursery Plant ◽  
Half Strength ◽  
Repeated Subculture

For high frequency regeneration of Rhyncnostylis retusa (Lin.) Blume apical nodal segments were used. Half strength MS + 2% sucrose + 1.5 mg/l BA + 0.5 mg/l NAA + 2 g/l peptone + 10% (v/v) coconut water (CW) + 0.5 g/l activated charcoal (AC) was the best nutrient medium, on which 89% cultures induced 8 microshoots per culture. Subculture of microshoots for further 8 weeks on the same nutrient medium enhanced the number of microshoots up to 95. For further proliferation of microshoots, their development into shoots as well as formation of secondary microshoots from the base of the old ones, the best medium was half strength of MS + 2% sucrose + 2 g/l peptone + 10% (v/v) CW + 0.5 g/l AC + 150 mg/l L-glutamine. Plantlets with roots were obtained in half strength of  MS + 2% sucrose + 2 g/l peptone + 10% (v/v) CW + 0.5 g/l AC + 5.0  g/l banana powder, on which cent per cent shoots rooted within eight weeks. The pH of all the categories of cultures were maintained at 5.6 before adding 2.2 g/l gelrite and autoclaving, and the cultures were incubated at 2000 - 3000 lux for 16/8 hrs light/dark at 24 ± 2ºC. Regeneration of plantlets continued due to repeated subculture of microshoots and regenerants were acclimatized and established in the nursery. Plant Tissue Cult. & Biotech. 22(1): 1-11, 2012  (June) DOI: http://dx.doi.org/10.3329/ptcb.v22i1.11242 

scholarly journals Clonal Micropropagation of representatives of the genus Dactylorhiza Neck. ex Nevski

2021 ◽  
Vol 4 (46) ◽  
pp. 17-17
Author(s):  
Alexander Saakian ◽  
◽  
Keyword(s):  
Survival Rate ◽  
Growth Regulators ◽  
In Vitro Propagation ◽  
Culture Medium ◽  
Culture Media ◽  
Ms Medium ◽  
Organic Additives ◽  
Clonal Micropropagation ◽  
Half Strength

Abstract The aim of this study is to develop and improve methods of in vitro propagation of representatives of Dactylorhiza: D.baltica , D. fuchsii. For the study, we used protocorms obtained by the asymbiotic germination of seed during 90 days. It has been established that half-strength of Murashige and Skoog (1962) medium (½ MS) supplemented with 1-2 mg/l 6-Benzylaminopurine(6-BAP), potato puree (20g/l), and charcoal (1g/l) effectively influenced the development of protocorms, and seedlings formation in the studied species. The result of the study showed that the survival rate of protocorms was high in all experimental culture media, but in D. fuchsii it was better at a concentration 2mg/l of 6-BAP (95.4%), while in D. baltica it was high at 1mg/l (87.0%). The highest percentage of multiple protocorms (68%) and the formation of new secondary protocorms in D. fuchsii (5,5±0,3 units) were observed on a culture medium containing 2 mg/l 6-BAP. The highest percent of rooting of D. fuchsii protosoms (78%) and length of roots (0.9cm) observed in ½ MS medium without growth regulators. During the development of D. baltica protosoms, the culture medium of ½ MS containing 1 mg/l 6-BAP had the best effect on the number of roots (1.8±0.1root/protosom), while the medium supplemented with 2mg/l of 6-BAP contributed to the formation of a larger number of new secondary protocorms (3,2±0,1protocorm/unit). During the subsequent cultivation of protosoms of D. baltica on a culture medium containing 1 mg/l it was observed an increase in the height of shoots (4,8±0,3 см), and the length of roots (2,2±0,1 см), wherein the number of newly formed protocorms was higher by 30% on the medium supplemented with 2 mg/l 6-BAP. Keywords: DACTYLORHIZA BALTICA, DACTYLORHIZA FUCHSII, IN VITRO, PROTOCORMS, ORGANIC ADDITIVES

scholarly journals Robust in vitro propagation and regeneration of ubi kuning high beta carotene cassava genotype through somatic embryogenesis

2017 ◽  
Vol 9 (4) ◽  
pp. 352-360
Author(s):  
SUPATMI SUPATMI ◽  
HANI FITRIANI ◽  
NURHAMIDARR RAHMAN ◽  
N. SRI HARTATI ◽  
ENNY SUDARMONOWATI
Keyword(s):  
Somatic Embryogenesis ◽  
Embryogenic Callus ◽  
Somatic Embryos ◽  
In Vitro Propagation ◽  
Light Condition ◽  
Beta Carotene ◽  
Induction Medium ◽  
Dark Light ◽  
High Beta

Supatmi, Fitriani H, Rahman N, Hartati NS, Sudarmonowati E. 2017. Robust in vitro propagation and regeneration of ubi kuning high beta carotene cassava genotype through somatic embryogenesis. Nusantara Bioscience 9: 352-360. Ubi kuning is a local genotype of cassava with high beta carotene content but the development of this genotype is still low because of plant disease susceptibility. Objectives of this study were to robust induce and regenerate somatic embryos of ubi kuning in vitro as well as to define a protocol of cyclic somatic embryogenesis of ubi kuning. Different size of leaf lobes, various concentration of picloram and different light conditions were tested to produce an effective and efficient somatic embryos (SEs).The best response of the induction of embryogenic callus was observed in leaf lobes explant with range size of 1-3 mm and >5mm cultured on induction medium (MS + 4% sucrose + 4 μM CuSO4 + 0.1 mM Glutamine + 0.8% Microagar) supplemented with either 10 or 18 mg/L picloram grown under dark light for 4 weeks. Retransferring embryogenic callus to the same medium supplemented with 16 mg/L picloram gave the advanced development of primary somatic embryos (PSEs) after 70 d grown under both dark and light condition treatments. A positive correlation between globular and cotyledon stages was obtained in all treatments (P≤ 0.01). The highest shoot and root growth (30% and 25%) was achieved in the regeneration of cotyledonary like-tissues cultured on callus embryogenic media (CEM) (MS basal+ 2.5 μM CuSO4 + 3% sucrose + 2.75 g/L phytagel) supplemented with 1.6 mg/L of BAP (6-Benzylaminopurine).

scholarly journals In vitro propagation of banana

1970 ◽  
Vol 34 (2) ◽  
pp. 269-278
Author(s):  
M Rezaul Karim ◽  
MA Malek ◽  
Sajia Rahman ◽  
M Al-Amin ◽  
M Ruhul Amin
Keyword(s):  
Plant Regeneration ◽  
In Vitro Propagation ◽  
Room Temperature ◽  
Ms Medium ◽  
Musa Sp ◽  
In Vitro Technique ◽  
Half Strength ◽  
The Mean ◽  
Basal Media

An in vitro technique for plant regeneration using meristem-derived plantlets of banana cv. BARI-l (Musa sp.) has been developed. Highest number of shoot regeneration was noticed on basal media supplemented with 7.5 mgL-1 BAP + 0.5 mgL-1 NAA at 30 days after inoculation (DAI). The mean number of shoots significantly reduced when the concentrations of BAP and NAA in the medium was high. Regenerated shoots were rooted on half strength MS medium containing 0.5 mgL-1 IAA + 0.5 mgL-1 IBA at 30 DAI. In vitro raised plantlets were transferred to poly bags containing ground soil and cowdung mixture (1:1) for acclimatization and hardening in room temperature (28-30°C) and the established plantlets are ready for planting in the field. Key Words: In vitro propagation; banana; Musa sp.DOI: 10.3329/bjar.v34i2.5799Bangladesh J. Agril. Res. 34(2): 269-278, June 2009

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