scholarly journals High Frequency Regeneration of Phalaenopsis amabilis (L.) Bl. cv. Lovely through In vitro Culture

1970 ◽  
Vol 20 (2) ◽  
pp. 185-193 ◽  
Author(s):  
Pinaki Sinha ◽  
Miskat Ara Akhter Jahan ◽  
John Liton Munshi ◽  
Rahima Khatun
Keyword(s):  
Plant Tissue ◽  
Activated Charcoal ◽  
High Frequency ◽  
Nutrient Medium ◽  
Clonal Propagation ◽  
Leaf Explants ◽  
Huge Amount ◽  
Leafy Shoots ◽  
Half Strength

Young leaf explants from mature plant of Phalaenopsis amabilis cv. Lovely were cultured on half strength of MS supplemented with BA (2.0 mg/l), NAA (0.5 mg/l), 2% (w/v) sucrose, 10% (v/v) coconut water, 2 g/l peptone and 1 g/l activated charcoal. Each section of explant pro­duced  ten protocorm-like bodies (PLBs) after 12 weeks of culture. The PLBs were subcultured on the same nutrient medium but without phytohormone and addition of 150 mg/l                     L-glutamine, where PLBs were found to be enlarged with leafy shoots and new PLBs were induced from the base of the old ones. Plantlet development from leafy shoots was achieved on half strength MS supplemented with 2 g/l peptone, 2% (w/v) sucrose, 10% (v/v) CW and 1 g/l activated charcoal, where 100% explants were developed into plantlets with roots within eight weeks. The addition of 2.5 g/l banana powder enhanced the number and length of roots. Within the first 32 weeks after initiation of culture about 1200 plantlets as well as a huge amount of PLBs were achieved from a single explant section. The plantlets were acclimated in natural environment.   Key words: Orchid, Phalaenopsis amabilis, Protocorm-like bodies, Clonal propagation   D.O.I. 10.3329/ptcb.v20i2.6913   Plant Tissue Cult. & Biotech. 20(2): 185-193, 2010 (December)

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 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 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 In vitro Propagation of Solanum capsicoides All. – An Important Therapeutic Agent 'Kantakari'

1970 ◽  
Vol 20 (2) ◽  
pp. 179-184
Author(s):  
N.P. Anish ◽  
M.G. Rajesh ◽  
Jiby Elias ◽  
N. Jayan
Keyword(s):  
Survival Rate ◽  
Key Words ◽  
Plant Tissue ◽  
In Vitro Propagation ◽  
Therapeutic Agent ◽  
Shoot Tip ◽  
Shoot Induction ◽  
Half Strength ◽  
Shoot Tip Explants

Shoot tip explants from in vitro germinated seedlings of Solanum capsicoides All. inoculated on MS containing 2 mg/l BA produced maximum shoot induction response (26 shoots per explant). Rooting of the microshoots (19.4 roots per explant) was obtained better in half strength of MS supplemented with NAA (0.5 mg/l). Well rooted plantlets were successfully hardened with 80 per cent survival rate.   Key words: Solanum capsicoides, Propagation, Therapeutic agent   D.O.I. 10.3329/ptcb.v20i2.6912   Plant Tissue Cult. & Biotech. 20(2): 179-184, 2010 (December)

scholarly journals IN VITRO REGENERATION AND MASS PROPAGATION OF AZIMA TETRACANTHA [LAM.] FROM THE LEAF EXPLANTS THROUGH CALLUS CULTURE

2017 ◽  
Vol 4 (2) ◽  
pp. 52-56
Author(s):  
Mallika Devi T
Keyword(s):  
Callus Induction ◽  
In Vitro Regeneration ◽  
Leaf Explants ◽  
Shoot Formation ◽  
Ms Medium ◽  
Apical Leaf ◽  
Half Strength ◽  
Regenerated Plantlets ◽  
Azima Tetracantha

In the present study the protocol for callus induction and regeneration in Azima tetracantha has been developed in culture medium. The young apical leaf explants were used for callus induction on MS medium containing BAP and NAA at 1.0 and 0.4mgl-1 respectively showed maximum callus induction (73%). The amount of callus responded for shoot formation (74%) was obtained in the MS medium containing BAP (1.5 mgl-1) and NAA (0.3mgl-1).The elongated shoots were rooted on half strength medium supplemented with IBA (1.5 mgl-1) and Kn (0.4 mgl-1) for shoots rooted. Regenerated plantlets were successfully acclimatized and hardened off inside the culture and then transferred to green house with better survival rate.

scholarly journals Development of an in vitro Callus Induction Protocol and Shoot Proliferation for Selected Jatropha (Jatropha curcas) Accessions

2020 ◽  
Vol 30 (1) ◽  
pp. 131-141
Author(s):  
Hundessa Fufa ◽  
Jiregna Daksa
Keyword(s):  
Shoot Regeneration ◽  
Callus Induction ◽  
Jatropha Curcas ◽  
Plant Tissue ◽  
Leaf Explants ◽  
Shoot Proliferation ◽  
Adventitious Shoot ◽  
Regeneration Medium ◽  
Adventitious Shoot Buds

The present study was undertaken to establish a protocol for in vitro callusing of three Jatropha accessions, namely Metema, Adami Tulu and Shewa Robit from leaf explants. The medium supplemented with combination of 4.44 μM BAP and 4.52 μM 2,4-D resulted in maximum percentage of callus (100%) formed for all accessions. The maximum shoot regeneration (66.67%) from callus with 10.13 number of shoot was obtained from Shewa Robit in MS medum fortified with TDZ (2.27 μM ) and IBA (0.49 μM ). The presence of TDZ in the shoot regeneration medium has greater influence on the induction of adventitious shoot buds, whereas MS supplemented with BAP alone and combination with IBA did not induce shoot regeneration from callus culture. The results obtained in the present study would facilitate the high callus induction and regeneration responses in Jatropha for its improvement using biotechnological tools. Plant Tissue Cult. & Biotech. 30(1): 131-141, 2020 (June)

scholarly journals Indirect in vitro Regeneration of Viola canescens Wall. ex, Roxb. by using Leaf Calli

2018 ◽  
Vol 28 (2) ◽  
pp. 215-222 ◽  
Author(s):  
Arun Kumar Khajuria ◽  
NS Bisht
Keyword(s):  
Plant Regeneration ◽  
Natural Population ◽  
Plant Tissue ◽  
In Vitro Regeneration ◽  
Soil Condition ◽  
Medicinal Herb ◽  
Growth Chamber ◽  
Half Strength ◽  
Number Of Shoots

An efficient indirect plant regeneration protocol was developed for Viola canescens, an important medicinal herb used in broad spectra of diseases in number of folk medicines since aeon. Excessive use of this plant without any rehabilitating measure has led to decline its natural population. Present investigation reports the use of zeatin to regenerate the plant from the callus on MS following its acclimatization on the soil condition. Calli of the plant responded positively to zeatin and maximum number of shoots 13.07 ± 2.01 were obtained when 9.12 μM concentration of zeatin was used. Regenerated shoots were subsequently rooted with IBA on MS and half strength MS and showed maximum number of roots 14.13 ± 1.64 after 60 days when medium was fortified with 4.92 μM IBA, followed by transferring them to soil condition, acclimatization of the plantlet was carried in growth chamber and then finally to the field for their survival where it showed 80% survival. Plant Tissue Cult. & Biotech. 28(2): 215-222, 2018 (December)

scholarly journals High-frequency Direct Somatic Embryogenesis and Plantlet Regeneration from Leaves Derived from In Vitro-Germinated Seedlings of a Coffea arabica Hybrid Cultivar

HortScience ◽  
2016 ◽  
Vol 51 (9) ◽  
pp. 1148-1152 ◽  
Author(s):  
Jane Kahia ◽  
Margaret Kirika ◽  
Hudson Lubabali ◽  
Sinclair Mantell
Keyword(s):  
Tissue Culture ◽  
High Frequency ◽  
Somatic Embryos ◽  
Coffea Arabica ◽  
Alternative Methods ◽  
Direct Somatic Embryogenesis ◽  
Ms Medium ◽  
Embryogenic Cultures ◽  
Half Strength

Breeding work carried out during the period 1971–85 by the Coffee Research Institute, Ruiru, Kenya resulted in the release of a new improved hybrid Coffea arabica named Ruiru 11. The cultivar combines resistance to coffee berry disease (CBD) and leaf rust, with high yield and good cup quality attributes. The propagation by F1 hybrid seeds production, cuttings, and tip grafting do not produce enough planting materials. There was a need to explore alternative methods and tissue culture offers potential options. The objective of the study was to evaluate the effect of explant sources and cytokinins on induction and regeneration of somatic embryos. Eight different explants were cultured on half-strength Murashige and Skoog (MS) medium supplemented with 10 µm benzylaminopurine (BAP). The effect of kinetin, N6-(2-isopentyl) adenine (2iP) evaluated at (0, 0.5, 5, or 25 µm) or thidiazuron (TDZ) (0, 0.5, 1.0, or 5 µm) added in separate experiments was also evaluated. The percentage of embryogenic cultures and the numbers of embryos per explant were determined after 3 months’ culture. The explant type had a significant effect (P > 0.05) on the induction of somatic embryos. Explants from in vitro-germinated seedlings produced the highest embryogenic cultures (90%) and the highest mean number of embryos (19.36) per explant. Cytokinins strongly enhanced induction and regeneration of somatic embryos. TDZ at 1 µm produced the highest embryogenic cultures (100%) and the highest mean number of embryos (24.2). The embryos were germinated on half-strength MS medium without any hormones. A high (98%) survival rate of the regenerated plantlets was recorded over all the treatments in the greenhouse. This is the first report on induction of high-frequency direct somatic embryos from coffee juvenile tissues. This is of great significance in tissue culture and indeed molecular biology manipulations because it allows regeneration of coffee from several explants.

scholarly journals Transformation of Nicotiana alata Link and Otto. with an Autoregulatory Senescence-inhibitor Gene Construct

HortScience ◽  
1996 ◽  
Vol 31 (4) ◽  
pp. 617c-617
Author(s):  
Kenneth R. Schroeder ◽  
Dennis P. Stimart
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Leaf Explants ◽  
Shoot Proliferation ◽  
Nicotiana Alata ◽  
Ms Medium ◽  
Gene Encoding ◽  
Half Strength ◽  
Cytokinin Synthesis ◽  
Senesced Leaves

Leaf explants of Nicotiana alata Link and Otto. were surface disinfested and cultured on Murashige and Skoog (MS) medium containing 2.66 μm N6-benzyladenine (BA) to promote shoot proliferation. After 5 weeks, proliferated shoots were removed and remaining callus saved. Callus was inoculated with Agrobacterium tumefaciens encoding a senescence-specific promoter SAG12 cloned from Arabidopsis thaliana fused to a Agrobacterium tumefaciens gene encoding isopentenyl transferase which catalyzes cytokinin synthesis. Following inoculation, the callus was cocultivated for 6 days on BA medium. Selection for transgenics was done on BA medium plus 100 mg Kanamycin and 400 mg Ticarcillin (antibiotics) per liter. Proliferating shoots were rooted on MS medium containing antibiotics. Rooted cuttings were transplanted to soil, acclimated and flowered in the greenhouse. Transgenics were outcrossed to a commercial N. alata hybrid. Seed was germinated in vitro on half-strength MS medium plus antibiotics. Segregation of transgenics to nontransgenics was 1:1. Evaluation of leaf senescence on 5-month-old plants showed 2 to 14 times fewer senesced leaves on the transgenic than the nontransgenic plants.

scholarly journals Isolated culture of A. reptance L., its’ morphological and growth features

2021 ◽  
Vol 40 ◽  
pp. 01001
Author(s):  
Elen Poghosyan ◽  
Naira Sahakyan ◽  
Margarit Petrosyan ◽  
Irina Batlutskaya ◽  
Karen Trchounian
Keyword(s):  
Callus Culture ◽  
Nutrient Medium ◽  
Leaf Explants ◽  
Shoot Formation ◽  
Callus Cultures ◽  
Soil Conditions ◽  
Naphthaleneacetic Acid ◽  
Micro Propagation ◽  
2,4 Dichlorophenoxyacetic Acid

A growing demand for the ecologically pure products brings us for searching novel biotechnological approaches for plant cultivation. One of these approaches is the in vitro cultivation and further acclimatization of valuable plant species. The object of our investigation was Ajugareptance L. ornamental plant which possesses high metabolic activity. In vitro cultivation was carried out applying Murashige-Skoog nutrient medium and its modifications. Acclimatization of in vitro plants was implemented according Hazarika. In the presence of twice higher concentration of cytokinins over auxins and 0.2 mg/ml gibberellins callus culture was formed from the leaf explants. Callus tissue was formed in the presence of 0.2 mg/ml kinetin and 2 mg/ml indole-3-acetic acid which has denser structure than the first one. The shoot formation was observed on callus cultures growing on the same medium approximately after 5th passage. Callus culture growth was supported also by the adding of 2 mg/ml 2,4-dichlorophenoxyacetic acid. For the micropropagation, the already formed shoots were transferred to the nutrient medium which contains only 0.1 mg/ml 1-Naphthaleneacetic acid as a phytohormone. A. reptans culture has high regenerative ability and the micro-propagation index was 104 – 105. In vitro regenerated plants were successfully acclimatized to the soil conditions during two-week period.

Sign in / Sign up

Export Citation Format

Share Document