scholarly journals Somatic embryogenesis from leaf transverse thin cell layer derived-callus of Vietnamese ginseng (Panax vietnamensis Ha et Grushv.)

2016 ◽  
Vol 14 (1) ◽  
pp. 63-73
Author(s):  
Vu Thi Hien ◽  
Nguyen Phuc Huy ◽  
Bui Van The Vinh ◽  
Hoang Xuan Chien ◽  
Hoang Thanh Tung ◽  
...  
Keyword(s):  
Somatic Embryogenesis ◽  
Cell Layer ◽  
Culture Media ◽  
Callus Formation ◽  
Basal Medium ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Transverse Thin Cell Layer ◽  
Cell Layers ◽  
2,4 Dichlorophenoxyacetic Acid

No report on plant regeneration via somatic embryogenesis of P. vietnamensis has been previously published. In the present study, somatic embryogenesis via callus formation from cultures of leaf transverse thin cell layers (tTCLs) of Vietnamese ginseng (Panax vietnamensis Ha et Grushv.) was investigated. α-naphthaleneacetic acid (NAA), 2,4-dichlorophenoxyacetic acid (2,4-D), 6-benzylaminopurine (BA) and thidiazuron (TDZ) were added separately and in combination into the culture media. Explant necrosis or low callogenesis rates were observed when 1-mm wide leaf tTCLs were cultured on media with TDZ, BA, 2,4-D or NAA. On the other hand, calli were successfully induced from the tTCL explants cultured on medium supplemented with either 2,4-D and BA or 2,4-D and TDZ. Callogenesis was observed under both light and dark conditions. The highest callogenesis rate (100%) was obtained on Murashige and Skoog (MS) basal medium supplemented with 1.0 mg l-1 2,4-D in combination with 0.1 mg l-1 TDZ in darkness after eight weeks of culture. White calli were cut into small pieces (1.0 x 1.0 cm dimension) and placed on MS media containing 1.0 mg l-1 2,4-D, 0.5 mg l-1 NAA and TDZ at various concentrations (0.01; 0.1; 0.2; and 0.5 mg l-1), and the best callus proliferation was recorded on medium containing 1.0 mg l-1 2,4-D and 0.2 mg l-1 TDZ. Somatic embryogenesis, with a success rate of 53.3% and 35 embryos per explant, was achieved when calli were subcultured onto MS medium supplemented with 1.0 mg l-1 2,4-D, 0.5 mg l-1 NAA and 0.2 mg l-1 TDZ.

Callus formation and shoot bud differentiation in anther culture of Solanum surattense

1979 ◽  
Vol 57 (22) ◽  
pp. 2524-2527 ◽  
Author(s):  
S. Sinha ◽  
R. P. Roy ◽  
K. K. Jha
Keyword(s):  
Anther Culture ◽  
Callus Formation ◽  
Low Frequency ◽  
Basal Medium ◽  
Pollen Grains ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Cytological Examination ◽  
Shoot Bud ◽  
2,4 Dichlorophenoxyacetic Acid

In anther culture of Solatium surattense, the Murashige and Skoog's medium supplemented with 2,4-dichlorophenoxyacetic acid (2.2 mg/L), indoleacetic acid or naphthaleneacetic acid (1.9 mg/L), and kinetin (2.2 mg/L) served as “callus-producing medium.” Histological and cytological observations indicated that the callus originated from the pollen grains. Synergistic action of kinetin (5.0 mg/L) and coconut milk (15%) in basal medium was able to induce differentiation of shoot buds either from the anthers directly or from the callus. Directly differentiating buds were formed by whole shoot bud morphogenesis of pollen. They were produced at a low frequency and showed presence of well-developed radicular and plumular regions. But the buds originating from callus lacked radicular ends. Root initiation in such buds was achieved by transferring them to basal medium. Cytological examination of the androgenic plantlets revealed a chromosomal series ranging from the haploid to the hexaploid with a few aneuploids.

scholarly journals Somatic embryogenesis and plant regeneration from callus cultures of Cleome rosea Vahl

2010 ◽  
Vol 53 (3) ◽  
pp. 679-686 ◽  
Author(s):  
Claudia Simões ◽  
Norma Albarello ◽  
Cátia Henriques Callado ◽  
Tatiana Carvalho de Castro ◽  
Elisabeth Mansur
Keyword(s):  
Somatic Embryogenesis ◽  
Callus Formation ◽  
Callus Cultures ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Stem Explants ◽  
Ms Medium ◽  
Fold Reduction ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Embryogenic Callus Formation

This paper describes a protocol for the efficient vegetative propagation of Cleome rosea by somatic embryogenesis. Leaf and stem explants from nursery-grown seedlings of C. rosea were cultivated on Murashige and Skoog (MS) medium supplemented with indole-3-acetic acid (IAA), a -naphthaleneacetic acid (NAA), 4-amino-3,5,6-trichloropicolinic acid (picloram) or 2,4-dichlorophenoxyacetic acid (2,4-D). Nodular calli were produced from both explant types in the presence of 4.5 and 9.0 µM 2,4-D. Embryo development and maturation were achieved when calli from stem explants were transferred to media containing a ten-fold reduction of 2,4-D concentration initially used (0.45 and 0.90 µM). Leaf-derived calli did not form embryos with the same treatments. The highest frequency of embryogenic callus formation (85%) and number of embryo per callus (13.45 ± 2.8) were achieved during the first subculture on medium supplemented with 0.90 µM 2,4-D. Embryo conversion into plantlets was achieved following transfer to growth regulator-free MS medium solidified with 2 g.L-1 phytagel. An acclimatization rate of 53% was found three months after transfer to ex vitro conditions and the recovered plants presented a normal phenotypic aspect.

Selectivity of auxin for induction and growth of callus from excised embryo of spring and winter wheat

1984 ◽  
Vol 62 (7) ◽  
pp. 1393-1397 ◽  
Author(s):  
M. D. Zhou ◽  
T. T. Lee
Keyword(s):  
Winter Wheat ◽  
Chinese Spring ◽  
Shoot Growth ◽  
Callus Formation ◽  
Triticum Aestivum L ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Methoxybenzoic Acid

The callus-promoting activity of most commonly known as well as some rarely tested auxins was compared with that of 2,4-dichlorophenoxyacetic acid (2,4-D) for in vitro culture of the excised embryo of spring and winter wheat (Triticum aestivum L.), cv. Chinese Spring and cv. Fredrick. Different auxins in a concentration range from 1 to 50 μM showed widely different activities. Also the two wheat cultivars responded differently to the auxins. When rapid callus formation with limited root growth was used as the basis for comparison, 2-(2-methyl-4-chlorophenoxy)propionic acid (2-MCPP), α-naphthaleneacetic acid, 3,6-dichloro-2-methoxybenzoic acid (dicamba), 4-amino-3,5,6,trichloropicolinic acid (picloram), γ-(2,4-dichlorophenoxy)butyric acid, 2,4,5-trichlorophenoxyacetic acid, and 2,4,5-trichlorophenoxypropionic acid, in the order of effectiveness, were superior to 2,4,-D for callus induction from the embryo of 'Chinese Spring,' although the concentration required was higher than that of 2,4-D. For the winter wheat 'Fredrick,' however, only picloram, dicamba, and 2-MCPP performed as well as 2,4-D. All auxins tested promoted shoot growth; 2-methyl-4-chlorophenoxypropionic acid was most effective for 'Chinese Spring,' whereas picloram was most effective for 'Fredrick.'

Regeneration of Robiniapseudoacacia via somatic embryogenesis

1989 ◽  
Vol 19 (2) ◽  
pp. 285-288 ◽  
Author(s):  
S. A. Merkle ◽  
A. T. Wiecko
Keyword(s):  
Somatic Embryogenesis ◽  
Somatic Embryos ◽  
Basal Medium ◽  
Direct Somatic Embryogenesis ◽  
Secondary Embryogenesis ◽  
Dichlorophenoxyacetic Acid ◽  
Entire Study ◽  
Fruit Maturity ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Free Media

Tissue cultures were initiated from developing seeds of black locust (Robiniapseudoacacia L.) collected from three trees at weekly intervals from 1 week following anthesis until early fruit maturity. Explants were cultured on media containing 0, 2, or 4 mg/L 2,4-dichlorophenoxyacetic acid and 0 or 0.25 mg/L 6-benzyladenine. Seeds explanted onto hormone-supplemented media remained on these media for 1 or 3 weeks before being placed on hormone-free media, or were maintained on hormone-supplemented media for the entire study. Direct somatic embryogenesis was observed in a single culture, initiated from a seed collected 4 weeks after anthesis and cultured for 1 week on a medium supplemented with 4 mg/L 2,4-dichlorophenoxyacetic acid and 0.25 mg/L 6-benzyladenine before transfer to basal medium. Although it could not be discerned from which part of the explant somatic embryos were derived, secondary embryogenesis continued from the radicles of cotyledonary-stage somatic embryos. Most somatic embryos were well formed, with two distinct cotyledons. Embryos germinated precociously, producing plantlets that were initially weak but later gained vigor and resembled seedlings.

scholarly journals Shoot Generation and Callus Induction of Dioscorea hispida Dennst by Different Plant Growth Hormones and Basal Media

2020 ◽  
Vol 11 (2) ◽  
pp. 30-38
Author(s):  
Nor Hasima Mahmod ◽  
Zakiah Mustapha ◽  
Ahmad Hilman Ariffin Husni ◽  
Nurul Anisah Ishak ◽  
Hafsah Jaafar
Keyword(s):  
Callus Culture ◽  
Basal Medium ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Plant Growth Hormones ◽  
Efficient Type ◽  
Medicinal Properties ◽  
Tuber Sprouting ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Stem Segments

Dioscorea hispida Dennst produces tuber which possess valuable medicinal properties but unsustainable harvesting has led to its reduction. The plant propagates slowly because of its low tuber sprouting rate. In average, Dioscorea hispida Dennst tubers took approximately 60 d to break dormancy and sprout. Hence, callus culture is proposed as a possible efficient type of culture for manipulation of this species.  In the present study, calli were induced from stem segments to evaluate callus culture potential of Dioscorea hispida Dennst. Results indicate that the combination of 1 mgL-1 naphthaleneacetic acid (NAA), 1 mgL-1 6- benzylaminopurine (BAP) and 0.5 mgL-1 2,4-dichlorophenoxyacetic acid (2, 4-D) in Gamborg (B5) medium improved callus multiplication and differentiation in the stem culture as opposed to those in Murashige and Skoog (MS) medium. The findings from the present study provide the basis of callus culture protocol for stem explant of Dioscorea hispida Dennst with B5 being the more effective basal medium.

scholarly journals Somatic Embryogenesis and Organogenesis in Magnolia dealbata Zucc. (Magnoliaceae), an Endangered, Endemic Mexican Species

HortScience ◽  
2006 ◽  
Vol 41 (5) ◽  
pp. 1325-1329 ◽  
Author(s):  
Martín Mata-Rosas ◽  
Ángel Jiménez-Rodríguez ◽  
Victor M. Chávez-Avila
Keyword(s):  
Somatic Embryogenesis ◽  
Basal Medium ◽  
Direct Somatic Embryogenesis ◽  
Direct Organogenesis ◽  
Limiting Factor ◽  
Zygotic Embryos ◽  
Dichlorophenoxyacetic Acid ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Magnolia Dealbata

Plants of Magnolia dealbata were regenerated from zygotic embryos through somatic embryogenesis and direct organogenesis. Medium and incubation conditions were determinating factors for the development of morphogenetic responses. Photoperiodic exposure was a limiting factor in the general development of the explants, and incubation in darkness allowed their development. The highest formation of shoots per responding explant were obtained on woody plant (WP) medium supplemented with 13.3 μM or 22.2 μM 6-benzylaminopurine (BA) in combination with 2.26 μM or in absence of 2,4-dichlorophenoxyacetic acid (2,4-D) from which 2.5 shoots per explant were induced. Subcultures on WP medium, supplemented with polyvinylpyrrolidone (PUP) 40,000 1 g·L–1) avoided necrosis of explants. Somatic embryos were formed in 85% of explants cultivated on WP medium with 2,4-D (2.3 μM or 4.5 μM); 20% induced indirect embryogenesis and 65% formed direct somatic embryogenesis. The plants were transferred to soil to acclimatize under greenhouse conditions, achieving 90% survival. Somatic embryo conversion to plantlets was obtained with subculture on WP basal medium without growth regulators. In vitro culture can play a key role in the propagation and conservation of this endangered species.

scholarly journals High Frequency Plant Regeneration System from Transverse Thin Cell Layer Section of In vitro Derived ‘Nadia’ Ginger Microrhizome

2014 ◽  
Vol 6 (1) ◽  
pp. 85-91
Author(s):  
Dikash Singh THINGBAIJAM ◽  
Devi Sunitibala HUIDROM
Keyword(s):  
High Frequency ◽  
Somatic Embryos ◽  
Cell Layer ◽  
Sucrose Concentration ◽  
Dichlorophenoxyacetic Acid ◽  
Thin Cell Layer ◽  
Regeneration System ◽  
Transverse Thin Cell Layer ◽  
2,4 Dichlorophenoxyacetic Acid

An efficient and reproducible procedure is outlined for rapid in vitro multiplication of Zingiber officinale var. ‘Nadia’ through high frequency shoot proliferation from transverse thin cell layer (tTCL) sections of in vitro derived microrhizome. In vitro derived microrhizome of size 500 μm in thickness was used as initial explants for induction of somatic embryos. Among the different phytohormones tested, tTCL explants shows maximum calli proliferation in medium containing 2 mg/L 2,4-Dichlorophenoxyacetic acid (88.30±0.11%). Reduced concentration of 2,4 Dichlorophenoxyacetic acid was supplemented with different cytokinins for regeneration of callus. Among the different medium tested, optimum redifferentiation of somatic embryos were observed in medium containing 0.2 mg/L 2,4 Dichlorophenoxyacetic acid and 6.0 mg/L BAP (141.08±0.25). Clump of regenerated plantlets were further subculture and transfer into microrhizome inducing medium containing high sucrose concentration (8%). Plantlets with well developed microrhizome were successfully acclimatized and eventually transferred to the field. The application of studying embryo section for regeneration of plants might be useful alternative to ginger improvement programme. Histological analysis showed formation of somatic embryos and regenerated adventitious shoot.

Isolation and cultivation of protoplasts from morphogenetic callus cultures of Lilium

1979 ◽  
Vol 57 (5) ◽  
pp. 512-516 ◽  
Author(s):  
John A. Simmonds ◽  
Daina H. Simmonds ◽  
Bruce G. Cumming
Keyword(s):  
Cell Wall ◽  
Culture Media ◽  
Continuous Light ◽  
Callus Cultures ◽  
Sodium Fluorescein ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Cell Wall Formation ◽  
Wall Formation ◽  
2,4 Dichlorophenoxyacetic Acid

Protoplasts isolated from Lilium callus which was maintained on media containing 2% sucrose contained large deposits of starch granules and lysed during isolation and washing procedures. Stable protoplast preparations could be obtained from callus which had been subcultured on sucrose-free medium for 3 weeks. Maximum protoplast yield (1.5 × 106 per gram fresh weight) was obtained when KCl (0.3 M) was the osmotic stabilizer. Inclusion of CaCl2 (25 mM) and MgSO4 (25 mM) in the isolation and wash media decreased protoplast lysis. Viability of protoplasts isolated in the high salts medium was determined by their ability to accumulate sodium fluorescein in the cytoplasm. No cell-wall formation occurred when salts were used as the osmoticum in various culture media. Continuous light (5000 lx) was inhibitory to protoplast survival. When protoplasts were transferred, via a series of wash solutions, to culture media using sugars as the osmoticum and cultured in darkness, cell-wall formation was detected after 3 days and cell divisions after 21 days. Zeatin (10−6 M), was needed for cell-wall formation. Cell division was stimulated by a combination of zeatin (10−6 M), naphthaleneacetic acid (10−5 M), and 2,4-dichlorophenoxyacetic acid (10−7 M) in the basic nutrient medium.

Embryo induction and plant regeneration of Callerya speciosa (Fabaceae) through anther culture

2017 ◽  
Vol 65 (1) ◽  
pp. 80 ◽  
Author(s):  
Bilan Huang ◽  
Li Xu ◽  
Kelie Li ◽  
Yunlu Fu ◽  
Zhiying Li
Keyword(s):  
Anther Culture ◽  
Culture Medium ◽  
Basal Medium ◽  
Dichlorophenoxyacetic Acid ◽  
Anther Wall ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Anther Cultures

An in vitro protocol for Callerya speciosa (Champ.) Schot regeneration through embryogenesis was developed using the anthers as the explants. The late uninucleate stage of the microspore was optimal for the anther culture of C. speciosa. Embryonic callus was induced on a MS basal medium supplemented with 4.4 µM 6-benzylaminopurine (BA) and 9.04 µM 2,4-dichlorophenoxyacetic acid (2,4-D). Embryos were obtained on MS medium supplemented with 2.2 µM BA and 0.5 µM naphthaleneacetic acid (NAA). The highest percentage (16.7%) of embryos was achieved using the culture medium MS + 0.25 µM NAA + 1.1 µM BA. The highest percentage of embryos that developed into plants was 18.3%. However, haploid plants were not observed, which may have been due to the collection of the calli from the anther wall. The results presented here demonstrate the establishment of a highly efficient and rapid system for regenerating C. speciosa using anther cultures.

scholarly journals The Effects of 2,4-Dichlorophenoxyacetic Acid and α-Naphthaleneacetic Acid on Biomass Increment, Rhizogenesis and Somatic Embryogenesis of Suspension-cultured Dinh Lang Cells [Polyscias fruticosa (L.) Harms]

2021 ◽  
Author(s):  
T.T.B. Phuong ◽  
V.P. Trung ◽  
N.H. An ◽  
N.D. Tuan ◽  
P.T.T. Nguyen
Keyword(s):  
Somatic Embryogenesis ◽  
Somatic Embryos ◽  
Bioactive Compound ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Fresh Weight ◽  
Triterpenoid Saponins ◽  
Cell Biomass ◽  
2,4 Dichlorophenoxyacetic Acid

Background: Dinh Lang [Polyscias fruticosa (L.) Harms] is a medicinal plant widely grown in Vietnam, with proven note-worthy health benefits. However, Dinh Lang’s amounts of triterpenoid saponins could not meet the need of the pharmaceutical industry. Thus, this study’s purpose is to figure out the optimal condition for raising Dinh Lang’s cell biomass, rhizogenesis and somatic embryogenesis to provide materials for bioactive compound productions. Methods: Different 2,4-dichlorophenoxyacetic acid and α-naphthaleneacetic acid concentrations (0.5, 1.0, 1.5 and 2.0 mg/L) were examined to determine the best amount of each plant growth regulator for raising cells’ biomass, rhizogenesis and somatic embryogenesis. In each treatment, two grams of eight-week-old calli were cultured in 50 mL of liquid MS medium. Result: It is demonstrated by the results that liquid MS medium containing 1.5 mg/L α-naphthaleneacetic acid has the capacity of producing the highest numbers of somatic embryos (489 embryos per flask) and rooted cells (259.5 cells per flask), while the fresh weight of cells cultured in the medium given 1.5 mg/L 2,4-dichlorophenoxyacetic acid reached its peak of 5.7 g.

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