scholarly journals In vitro Embryogenesis Derived from Shoot Tips in Mass Propagation of Two Selected-Clones of Phalaenopsis

2016 ◽  
Vol 8 (3) ◽  
pp. 317-325 ◽  
Author(s):  
Budi WINARTO ◽  
Kristina Dwi ATMINI ◽  
Dedeh Siti BADRIAH ◽  
Mega WEGADARA
Keyword(s):  
Economic Value ◽  
Shoot Tips ◽  
Ms Medium ◽  
Market Demand ◽  
Mass Propagation ◽  
Full Strength ◽  
Half Strength ◽  
Alternative Choice ◽  
In Vitro Embryogenesis

Phalaenopsis is of high economic value and market demand in Indonesia; however, orchid products are mostly imported from other countries. ‘Kristina Dwi’ (KD) 69.274 and ‘Dedeh’ (D) 802.28 are two selected clones with high potential utilized and developed commercially. To support their commercialization, a reliable in vitro propagation protocol is essential.  In the current study, an in vitro mass propagation protocol for KD 69.274 and D 802.28 clones was successfully established using shoot tips as explant sources. A high number of embryos, up to 8.2 embryos per explant, with 58.5% explant regeneration, and 3.5 regenerated-explants in average were regenerated from shoot tips of KD 69.274 clone cultured on  half-strength Murashige and Skoog (MS) medium, with full strength micro, Fe-chellate and vitamin containing 0.5 mg/L thidiazuruon (TDZ) and 0.25 mg/L N6-benzyladenine (BA). The initial embryos were proliferated by culturing embryos individually on half-strength MS medium with 0.13 mg/L TDZ and 0.25 mg/L BA and resulted in high embryo regeneration up to 91.4%, with 10.2 embryos per explant and no embryo browning. The embryos were multiplied under periodical subcultures of 3 months each, resulting in gradual increasing number of embryos from the first subculture till the fifth subculture, with 23.6 embryos produced, then declined afterward. The embryos were easily germinated on half-strength MS medium with full strength of vitamin and hormone free, with 73.9% embryo germination and 14.9 germinated embryos. Healthy plantlets were stimulated on the same medium with 2 g/L activated charcoal (AC) and successfully acclimatized on Cycas rumphii bulk, with 88.3% survival plantlets. Finally, it can be summarized that a new in vitro mass propagation protocol, as new alternative choice for Phalaenopsis propagation, was successfully established.

scholarly journals Mass Propagation of Feronia limonia L. through Tissue Culture

1970 ◽  
Vol 45 (1) ◽  
pp. 75-78 ◽  
Author(s):  
Shahina Islam ◽  
Mosfequa Zahan ◽  
Shahina Akter ◽  
Tanjina Akhtar Banu ◽  
Ahashan Habib ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Plant Growth ◽  
Key Words ◽  
Multiple Shoot ◽  
Shoot Tips ◽  
Nodal Explants ◽  
Ms Medium ◽  
Ex Vitro ◽  
Mass Propagation

An efficient mass propagation method for Feronia limonia was developed from excised shoot tips and nodal explants of in vitro grown seedlings. Explants were cultured on MS medium with different conc. of NAA, Kn, IAA and BAP singly or in combinations. Highest number of micro shoots and better plant growth were obtained from these two explants on MS medium supplemented with 0.2 mg/l BAP alone. The regenerated shoots were successfully rooted on MS medium supplemented with 0.5 mg/l NAA. The in vitro raised plantlets were successfully established in soil following the formation of roots with 100% survivability under ex vitro condition. Key words: Feronia limonia; Mass propagation; Node; Shoot tips; Multiple shoot DOI: 10.3329/bjsir.v45i1.5186 Bangladesh J. Sci. Ind. Res. 45(1), 75-78, 2010

scholarly journals Regeneration of Hemidesmus indicus (L.) R. Br. using in vitro nodes: an alternative method for efficient multiplication of shoots

2021 ◽  
Vol 13 (2) ◽  
pp. 10831
Author(s):  
Ashutosh R. PATHAK ◽  
Aruna G. JOSHI
Keyword(s):  
Multiple Shoots ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Nodal Explant ◽  
Mass Propagation ◽  
Hemidesmus Indicus ◽  
Full Strength ◽  
Alternative Method

In vivo nodes of Hemidesmus indicus (L.) R. Br. induced healthy multiple shoots with branching in our earlier studies and thus in the present study, potency of in vitro nodes to regenerate shoots was evaluated. In vitro nodes were excised from eight-week-old shoots and placed in Murashige and Skoog’s (MS) medium fortified with sucrose (3%) and different concentrations of 6-benzyladenine (BA) and kinetin (Kn). After eight weeks, optimum of 5.42 ± 0.36 shoots with 100% response were regenerated in medium supplemented with BA (10 µM) and Kn (5 µM). These healthy shoots were placed in full, half and quarter strengths of liquid MS medium fortified with sucrose (1%) and α-naphthaleneacetic acid (NAA, 1-25 µM) for rooting. Among all the strengths of MS medium, full strength MS medium having 8 µM NAA formed maximum of 3.42 ± 0.55 roots (91.67% response) within four weeks. The protocol is in continuation with earlier study and it was confirmed that a single in vivo nodal explant can regenerate around 385 healthy elongated shoots within 4 months, which will help in mass-propagation of the species.

scholarly journals Direct Organogenesis from Rhizome Explants in Marsilea quadrifolia L.: A Threatened Fern Species

2015 ◽  
Vol 2015 ◽  
pp. 1-6 ◽  
Author(s):  
Mahipal S. Shekhawat ◽  
M. Manokari
Keyword(s):  
Multiple Shoots ◽  
Direct Organogenesis ◽  
Root Induction ◽  
Ms Medium ◽  
Full Strength ◽  
Marsilea Quadrifolia ◽  
Fern Species ◽  
Half Strength

An efficient micropropagation protocol has been developed for Marsilea quadrifolia L. through direct organogenesis. The mature rhizomes were used as explants and successfully sterilized using 0.1% HgCl2 for the establishment of cultures. The multiple shoots were differentiated from the explants on Murashige and Skoog (MS) medium augmented with 6-benzylaminopurin (BAP). Full strength MS medium was reported to be effective for the induction of sporophytes from the rhizomes after four weeks of inoculation. Maximum response (96%) with average of 6.2 shoots (2.72 cm length) was achieved on full strength of MS medium augmented with 0.5 mg/L BAP in culture initiation experiments. The cultures were further proliferated in clusters (79.0±0.37 shoots per explant) with stunted growth on half strength MS medium supplemented with 0.25 mg/L BAP after four weeks. These stunted shoots were elongated (5.30 cm long) on half MS medium devoid of growth hormones. Root induction and proliferation (3.0–4.0 cm long) were observed after 4th subculture of sporophytes on hormone-free half strength MS medium. The rooted plantlets were hardened in the fern house for 4-5 weeks and transferred to the field with 92% survival rate. There were no observable differences in between in vivo grown and in vitro propagated plantlets in the field.

scholarly journals Axillary Shoots Derived from Thin Cell Layer and Adenine Sulphate Application in in vitro Mass Propagation of Gerbera [Gerbera jamesonii (H. Bolus ex Bolus f.)]

2019 ◽  
Vol 11 (1) ◽  
pp. 63-76
Author(s):  
Budi WINARTO ◽  
Kurnia YUNIARTO ◽  
Dan M. WEGADARA
Keyword(s):  
Optimal Combination ◽  
Shoot Tip ◽  
Basic Medium ◽  
Ms Medium ◽  
Gerbera Jamesonii ◽  
Mass Propagation ◽  
Adenine Sulphate ◽  
Half Strength ◽  
Shoot Tip Explants

A new route of in vitro mass propagation protocol of Gerbera jamesonii (H. Bolus ex Bolus f.) derived from application of thin cell layers (TCL) and adenine sulphate (AS) was successfully developed and established. Shoot tip explants and half-strength MS medium containing 0.25 mg/l N6-benzylaminopurine (BAP), 20 g/l sucrose and 7 g/l Swallow agar were used as explant source and basic medium. Different TCL of transversal TCL (tTCL) and longitudinal TCL (lTCL) in four slicing positions of 1, 2, 3 and 4; varieties and clones i.e. G. jamesonii ‘Black Jack’, ‘Carambole’, ‘Nuance’, ‘Violente’, 01.098 and 11.46 clone; AS concentrations viz. 0, 20, 40, 60, 80 and 100 mg/l were tested in the study. Each step of in vitro culture established had unique and specific results. In the initiation stage, first slicing position of ‘Black Jack’ shoot tip tTCL was the most optimal combination treatment to produce 7.0 shoots per explant with 13.5 leaves. The first slicing position on shoot tip explants of 01.098 clone tTCL and 20 mg/l AS in half-strength MS medium containing 0.25 mg/l BAP were the most optimal combination treatment in obtaining the highest number of shoots produced per shoot up to 9.4 shoots per shoot with 34.1 leaves and 2.37 cm length of leaves in the proliferation stage, however the treatment did not give significant effect compared to control. Under periodical subcultures on the basic medium, number of shoots and leaves increased gradually from the initial culture with 3-6 shoots per shoot and 9.4-11.6 leaves till the fourth subculture with 6-11 shoots per shoot and 16.7-28.8 leaves and declined thereafter. Subculturing of shoots in accordance to produce qualified shoots for planting materials could be carried out till sixth to seventh subculture. The highest shoot multiplication rate (SMR) was established on 01.098 clone with as high as 7.3. The well shoots were easily rooted on half-strength MS medium supplemented with 0.1 mg/l BAP, 0.05 mg/l NAA and 1.5 g/l AC. Plantlets were then transferred to ex vitro condition for acclimatization on a mixture of burned-rice husk and organic manure (1:1, v/v) with 85-100% survivability. The ‘Black Jack’ and 11.46 clone were the best genotypes on the acclimatization stage with 100% survivability of plantlets. Results of the study have implication that first slicing position of shoot tip tTCL can be applied in establishing of in vitro propagation protocol for other gerberas.

scholarly journals Mass propagation of Bambusa bambos (L.) Voss through in vitro culture

2017 ◽  
Vol 5 (2) ◽  
pp. 15-26 ◽  
Author(s):  
Raihan I Raju ◽  
Shyamal K Roy
Keyword(s):  
In Vitro Culture ◽  
Basal Medium ◽  
Nodal Segments ◽  
Garden Soil ◽  
Root Induction ◽  
Ms Medium ◽  
Mass Propagation ◽  
Surface Sterilization ◽  
Half Strength

Protocol for mass propagation of Bambusa bamboos (L.) Voss was developed through in vitro culture. Nodal segments containing pre-existing axillary bud, after surface sterilization, were inoculated on liquid Murashige and Skoog’s (MS) basal medium containing different concentrations and combinations of cytokinins (BAP, TDZ and Kn). The highest direct shoot induction (90%) was obtained in the MS liquid medium supplemented with 2.0 mg/l BAP and 1.0 mg/l TDZ with maximum average number of shoots (3.14 ± 0.06) per explant. Highest shoot multiplication (16.58 ± 0.24 shoots per culture) with highest average shoot length (9.21 ± 0.13 cm) was obtained when in vitro raised shoots were cultured in gelrite gelled MS medium in conjunction with 2.0 mg/l BAP and 1.0 mg/l TDZ. Incorporation of 10% coconut water with 4% sucrose in the above mentioned medium resulted satisfactory shoot growth and development with an average 26.7 ± 0.60 shoots per culture. For root induction, in vitro raised shoots were divided into clumps of 4-5 shoots in each clump and transferred onto both liquid and gelled half-strength MS medium containing different concentrations and combinations of auxins (IBA and NAA). Maximum rooting (86.67%) was achieved in half-strength of MS medium fortified with 2.5 mg/l IBA and 2.5 mg/l NAA with an average 8.72 ± 0.42 root per shoot. The rooted plantlets were then transferred to polybags containing garden soil, sand and compost mixture with 1:1:1 ratio. After a month the hardened plantlets were then transferred to the larger pots containing garden soil and compost with 1:1 ratio for sufficient growth and finally transplanted to the field. In this process, the highest 100% survivability was recorded from well-established rooted plantlets. The regenerated plants showed well developed root and shoot systems in field condition.Jahangirnagar University J. Biol. Sci. 5(2): 15-26, 2016 (December)

scholarly journals Regeneration of Ficus glomerata Roxb., using shoot tips and nodal explants

1970 ◽  
Vol 39 (1) ◽  
pp. 47-50 ◽  
Author(s):  
AKM Sayeed Hassan ◽  
Rahima Khatun
Keyword(s):  
Survival Rate ◽  
Shoot Proliferation ◽  
Shoot Tips ◽  
Nodal Explants ◽  
Ms Medium ◽  
Shoot Induction ◽  
Half Strength ◽  
Regenerated Plantlets ◽  
Ficus Glomerata

Shoot tips and nodal explants from in vitro growing seedlings of Ficus glomerata Roxb. (Moraceae). showed best shoot induction (88%) on MS medium supplemented with 0.5 mg/l BAP, where maximum number of shoots were produced per culture. In vitro raised shoots rooted well on half strength of MS medium with 2.0 mg/l IBA + 0.1 mg/l NAA. The survival rate of regenerated plantlets was 82%. Key words: Ficus glomerata Roxb.; Shoot proliferation; Micropropagation; Acclimatization DOI: 10.3329/bjb.v39i1.5525Bangladesh J. Bot. 39(1): 47-50, 2010 (June)

scholarly journals In vitro regeneration for mass propagation in commercial scale of medicinal plant vitex nigundo L.

1970 ◽  
Vol 18 ◽  
pp. 140-145 ◽  
Author(s):  
Md Abu Hena Mostofa Jamal ◽  
ANM Rubaiyath-Bin Rahman ◽  
Dipak Kumar Paul ◽  
Md Rezuanul Islam
Keyword(s):  
Survival Rate ◽  
Medicinal Plant ◽  
Shoot Tips ◽  
Nodal Segments ◽  
Root Induction ◽  
Ms Medium ◽  
Shoot Induction ◽  
Mass Propagation ◽  
Commercial Scale

Context: It is necessary to focus on the importance of adopting micropropagation technique for mass propagation of the plantlets in commercial scale as well as conservation and distribution of germplasm. Objective: The present investigation has been designed with a view to establishing protocol of in vitro regeneration of medicinal plant species i,e., Vitex nigundo L (Verbenaceae). Materials and Methods: Shoot tips and nodal segments were used for multiple shoot induction. All explants were cultured on MS medium supplemented with various plant growth regulators. HgCl2 was used as surface sterilizing agent. For in vitro rooting, individual shoots (3-4 cm) were cut from the proliferated shoot cultures and implanted on half and full strength of MS with different concentrations and combinations of NAA and IAA. The cultures were incubated for 16 h photoperiod at 25 ± 2ºC under a fluorescent light. Visual observation of culture was made every week. Data on shoot induction and proliferation and root induction were recorded after three weeks of inoculation and used for calculation. For each treatment 15 explants were used and all the treatments were repeated thrice. Established plantlets were transplanted in earthen pots under natural conditions and the survival rate was recorded. Results: The most effective surface sterilization treatment has been found 0.1 % HgCl2 for 7 minutes. Highest number of shoot was observed in MS medium supplemented with 3.0 mg/ BAP. It was rooted well in full MS containing 2.0 mg/l IAA. The survival rate was 85 % and propagated plantlets were successfully acclimatized in soil. Conclusion: It was observed that shoot tips are more responsive for micropropagation of Vitex nigundo L . Thus the fruitful utilization of rapid clonal propagation, germplasm conservation and distribution of Vitex nigundo, important medicinal plant of Bangladesh, is possible. Keywords: Vitex nigundo; Medicinal plant; Shoot induction; Micropropagation; Regeneration. DOI: http://dx.doi.org/10.3329/jbs.v18i0.8790 JBS 2010; 18(0): 140-145

scholarly journals Nutritional requirements for germination and in vitro development of three Orchidaceceae species in the southern Brazilian Amazon

2018 ◽  
Vol 24 (2) ◽  
pp. 87-94 ◽  
Author(s):  
Viviane Luiza Hunhoff ◽  
Lais Alves Lage ◽  
Ednamar Gabriela Palú ◽  
Willian Krause ◽  
Celice Alexandre Silva
Keyword(s):  
Activated Charcoal ◽  
Culture Media ◽  
Leaf Number ◽  
Alternative Form ◽  
Ms Medium ◽  
Root Number ◽  
Shoot Height ◽  
Full Strength ◽  
Half Strength

Tissue culture is an alternative form of producing healthy, vigorous and regular plants on a large scale. The purpose of this study was to evaluate the most efficient culture medium for in vitro plantlet germination and development of three Orchidaceae species. Seeds disinfested of three species were dispersed in distilled water and dripped into basic Murashige and Skoog (MS) medium. The experimental design was completely randomized in a factorial 3 x 4 (three species x four culture media), with 5 replications. Four treatments were established: (1) full-strength MS medium, with the full nutrient concentration (MSØ), (2) full-strength MS medium plus 0.3% activated charcoal (MSØ ACh), (3) half- strength MS medium (½ MS) and (4) half- strength MS medium with 0.3 % activated charcoal (½ MS ACh). Germination was evaluated after 15, 20, 25, 30, and 60 days. The shoot height, leaf number and length, root number and length of plantlets of the three studied species were assessed. In A. variegata, 73% germinated after 60 days in ½ MS ACh medium. In the same period, 100% of E. viparum and S. gloriosa seeds germinated in MSØ ACh medium. The plant height, leaf number and length, root number and length were significantly higher for the species A. variegata and E. viviparum in MSØ ACh medium. The culture media ½ MS and MSØ with addition of activated charcoal favored in vitro germination for the three orchid species of this study.

scholarly journals In Vitro Secondary Embryogenesis Derived from Meta-Topoline Treatment on Mass Propagation of Phalaenopsis ‘AMP 17’

2016 ◽  
Vol 8 (1) ◽  
pp. 62-68
Author(s):  
Dewi PRAMANIK ◽  
Muhammad PRAMA YUFDY ◽  
Herni SHINTIAVIRA ◽  
Budi WINARTO
Keyword(s):  
Secondary Embryogenesis ◽  
Multiplication Rate ◽  
Market Demand ◽  
Mass Propagation ◽  
Flower Stalk ◽  
High Productivity ◽  
The Third ◽  
Secondary Somatic Embryo ◽  
Half Strength

Phalaenopsis ‘AMP 17’ is an important orchid commodity in Indonesia with high market demand; however, scaling up the orchid commercially is constrained by the availability and sustainability of qualified seedlings. To overcome the problem, a reliable in vitro propagation protocol, especially via secondary embryogenesis, was undertaken. In the present study, in vitro secondary embryogenesis derived from meta-topoline (mT) treatment on mass propagation of Phalaenopsis ‘AMP 17’ was successfully established. Embryos, as explant sources, were prepared by culturing meristem tips of flower stalk shoots on Murashige and Skoog (MS) medium containing 1.5 mg/L thidiazuron (TDZ) and 0.25 mg/L N6-benzylaminopurine (BAP) for ± 3 months. High secondary somatic embryo (SSE) formation up to 64.90% with 12.30 SSEs regenerated per embryo was determined on half-strength MS augmented with 0.5 mg/L BAP and 2.5 mg/L mT. The combination also stimulated the result of high multiplication rate of SSE formation, up to 10.1 fold on the third subculture, maintained low conversion rate of germinated-embryos down to 55% and improved qualified-growth of the germinated embryos. The mT treatment produced 86% survival plantlets with high qualified-performance. The system could be applied as an alternative method to step forward towards an improved propagation protocol, commercially efficient due to high productivity. Detail findings in each step were discussed.

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.

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