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 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 In vitro Mass Clonal Propagation of Spathoglottis plicata Blume

1970 ◽  
Vol 19 (2) ◽  
pp. 151-160 ◽  
Author(s):  
Pinaki Sinha ◽  
M. Lokman Hakim ◽  
M. Firoz Alam
Keyword(s):  
Clonal Propagation ◽  
Nodal Segments ◽  
Fluorescent Light ◽  
Plantlet Formation ◽  
The Third ◽  
Regenerated Plants ◽  
Rooting Medium ◽  
Half Strength ◽  
In Vitro Clonal Propagation

For in vitro clonal propagation of Spathoglottis plicata Blume nodal segments of young shoots were cultured on half strength of MS  with  2% sucrose + 2.0 mg/l BA + 0.5 mg/l NAA + 2 g/l peptone + 15% (v/v) CW + 0.5 g/l AC,  incubated at 24 ± 2ºC under 3000 lux fluorescent light for a 16 hr photoperiod per day. About 19 micro-shoots were induced from the explants within 12 weeks. Subculture of micro-shoots for eight weeks on the same nutrient medium enhanced the number of micro-shoots up to 60. The clumps of the micro-shoots were dissected and cultured on half strength of MS  with 2% sucrose + 2 g/l peptone + 15% (v/v) CW + 0.5 g/l AC + 200 mg/l L-glutamine. The micro-shoot sections elongated to form shoots, and new micro shoots were induced from the base within eight weeks of culture. For plantlet formation the best rooting medium was determined as  half strength of MS  with 2% sucrose + 2 g/l peptone + 15% (v/v) CW + 0.5 g/l AC + 50 g/l banana pulp. After rearing 25 g mixture of urea, TSP and MOP (2 : 1 : 1) were applied per plant at three months intervals. All the regenerated plants blossomed on the third year. Key words: Spathoglottis plicata, Clonal propagation, Acclimation D.O.I. 10.3329/ptcb.v19i2.5432 Plant Tissue Cult. & Biotech. 19(2): 151-160, 2009 (December)

scholarly journals In Vitro Regeneration and Mass Propagation of Ruta graveolens L.—A Multipurpose Shrub

HortScience ◽  
2005 ◽  
Vol 40 (5) ◽  
pp. 1478-1480 ◽  
Author(s):  
Mohd Faisal ◽  
Naseem Ahmad ◽  
Mohammad Anis
Keyword(s):  
High Frequency ◽  
In Vitro Regeneration ◽  
Nodal Segments ◽  
Ms Medium ◽  
Ruta Graveolens ◽  
Mass Propagation ◽  
Regeneration Frequency ◽  
Whole Plant ◽  
Half Strength

A protocol for rapid in vitro propagation of Ruta graveolens L. through high-frequency shoot induction from nodal explants was established. Proliferation of shoots from nodal segments was achieved on Murashige and Skoog medium supplemented with various concentrations of BA, Kin, IAA, and NAA, either singly or in various combinations. The highest shoot regeneration frequency (98.5%) and the highest number of shoots per explant (40.2 ± 2.8) was obtained on MS medium supplemented with 10 μm BA and 2.5 μm NAA. In vitro regenerated shoots rooted best on half-strength MS medium containing 0.5 μm IBA. Rooted shoots, following acclimatization in the greenhouse, were successfully transferred to field conditions, and 90% of plants survived. The efficient in vitro regeneration of the whole plant can be used as a fast and reliable method to transform R. graveolens genetically for its active principles.

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 multiplication, micromorphological studies and ex vitro rooting of Hybanthus enneaspermus (L.) F. Muell. – a rare medicinal plant

2018 ◽  
Vol 77 (1) ◽  
pp. 80-87 ◽  
Author(s):  
Mahipal S. Shekhawat ◽  
M. Manokari
Keyword(s):  
Medicinal Plant ◽  
Large Scale ◽  
Nodal Segments ◽  
Ms Medium ◽  
Ex Vitro ◽  
Culture Bottle ◽  
Hybanthus Enneaspermus ◽  
Half Strength

AbstractHybanthus enneaspermusis a rare medicinal plant. We defined a protocol for micropropagation,ex vitrorooting of cloned shoots and their acclimatization. Surface-sterilized nodal segments were cultured on Murashige and Skoog (MS) medium with different concentrations of 6-benzylaminopurine (BAP) and kinetin (Kin). Medium supplemented with 1.5 mg L−1BAP was found optimum for shoot induction from the explants and 6.4±0.69 shoots were regenerated from each node with 97% response. Shoots were further proliferated maximally (228±10.3 shoots per culture bottle with 7.5±0.43 cm length) on MS medium augmented with 1.0 mg L−1each of BAP and Kin within 4–5 weeks. The shoots were rootedin vitroon half strength MS medium containing 2.0 mg L−1indole-3 butyric acid (IBA). The cloned shoots were pulse-treated with 300 mg L–1 of IBA and cultured on soilrite® in a greenhouse. About 96% of the IBA-pulsed shoots rootedex vitroin soilrite®, each shoot producing 12.5±0.54 roots with 5.1±0.62 cm length. Theex vitrorooted plantlets showed a better rate of survival (92%) in a field study thanin vitrorooted plantlets (86%). A comparative foliar micromorphological study ofH. enneaspermuswas conducted to understand the micromorphological changes during plant developmental processes fromin vitrotoin vivoconditions in terms of variations in stomata, vein structures and spacing, and trichomes. This is the first report onex vitrorooting inH. enneaspermusand the protocol can be exploited for conservation and large-scale propagation of this rare and medicinally important plant.

scholarly journals Influence of Cytokinins in Combination with GA3on Shoot Multiplication and Elongation of Tea Clone Iran 100 (Camellia sinensis(L.) O. Kuntze)

2014 ◽  
Vol 2014 ◽  
pp. 1-9 ◽  
Author(s):  
Reza Azadi Gonbad ◽  
Uma Rani Sinniah ◽  
Maheran Abdul Aziz ◽  
Rosfarizan Mohamad
Keyword(s):  
Camellia Sinensis ◽  
Woody Plants ◽  
Clonal Propagation ◽  
Shoot Multiplication ◽  
Shoot Elongation ◽  
Nodal Segments ◽  
Nodal Segment ◽  
Liquid Form ◽  
Solid Media

The use ofin vitroculture has been accepted as an efficient technique for clonal propagation of many woody plants. In the present research, we report the results of a number of experiments aimed at optimizing micropropagation protocol for tea (Camellia sinensis(L.) O. Kuntze) (clone Iran 100) using nodal segments as the explant. The effect of different combinations and concentrations of plant growth regulators (PGR) (BAP, TDZ, GA3) on shoot multiplication and elongation was assessed. The influence of exposure to IBA in liquid form prior to transfer to solid media on rooting of tea microshoots was investigated. The results of this study showed that the best treatment for nodal segment multiplication in terms of the number of shoot per explant and shoot elongation was obtained using 3 mg/L BAP in combination with 0.5 mg/L GA3. TDZ was found to be inappropriate for multiplication of tea clone Iran 100 as it resulted in hyperhydricity especially at concentrations higher than 0.05 mg/L. Healthy shoots treated with 300 mg/L IBA for 30 min followed by transfer to 1/2 strength MS medium devoid of PGR resulted in 72.3% of shoots producing roots and upon transferring them to acclimatization chamber 65% survival was obtained prior to field transfer.

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 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.

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