SOMATIC EMBRYOGENESIS IN CELL SUSPENSION CULTURE OF COWPEA (VIGNA UNGUICULATA (L.) WALP)

1995 ◽  
Vol 43 (4) ◽  
pp. 385-390 ◽  
Author(s):  
S. Kulothungan ◽  
A. Ganapathi ◽  
A. Shajahan ◽  
K. Kathiravan
Keyword(s):  
Somatic Embryogenesis ◽  
Liquid Medium ◽  
Vigna Unguiculata ◽  
Somatic Embryos ◽  
Leaf Explants ◽  
Dichlorophenoxyacetic Acid ◽  
Ms Medium ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Seedling Leaf ◽  
Cotyledonary Stage

Embryogenic callus was induced from seedling leaf explants of cowpea (Vigna unguiculata (L.) Walp. cv. C152 on Murashige and Skoog (MS) medium containing 2.0 mg 1−1 2,4-dichlorophenoxyacetic acid (2,4-D). The maximum frequency of somatic embryogenesis was noticed when this callus was transferred to MS liquid medium supplemented with 2 mg 1−1 2,4-D. Further studies on ontogeny of somatic embryos showed that the cells destined to become somatic embryos divided into spherical or filamentous proembryos. Subsequent divisions in the proembryo led to globular, heart, torpedo-shaped, and cotyledonary-stage somatic embryos. Tiny plantlets were obtained by transferring the cotyledonary-stage somatic embryos to MS liquid medium containing 0.5 mg 1−1 2,4-D.

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.

scholarly journals Plant regeneration from protoplasts of Gentiana straminea Maxim

2016 ◽  
Vol 11 (1) ◽  
pp. 55-60 ◽  
Author(s):  
Guomin Shi ◽  
Lina Yang ◽  
Tao He
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Somatic Embryos ◽  
Protoplast Culture ◽  
Dichlorophenoxyacetic Acid ◽  
Ms Medium ◽  
Fresh Weight ◽  
Embryogenic Calli ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Solid Liquid

AbstractA protocol is described for plant regeneration from protoplasts of Gentiana straminea Maxim. via somatic embryogenesis. Protoplasts were isolated from embryogenic calli in an enzyme solution composed of 2% Cellulase Onozuka R-10, 0.5% Macerozyme R-10, 0.5% Hemicellulase, and 0.5 M sorbitol with a yield of 3.0 × 106 protoplasts per gram of fresh weight. Liquid, solid-liquid double layer (sLD) and agar-pool (aPL) culture systems were used for protoplast culture. The aPL culture was the only method that produced embryogenic, regenerative calli. With aPL culture, the highest frequencies of protoplast cell division and colony formation were 39.6% and 16.9%, respectively, on MS medium supplemented with 2 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.5 mg/L N6-benzylaminopurine (BA). Microcalli were transferred to solid MS medium containing a reduced concentration of 2,4-D (0.5 mg/L) to promote the formation of embryogenic calli. Somatic embryos developed into plantlets on MS medium supplemented with 2 mg/L BA at a rate of 43.7%.

scholarly journals Somatic Embryogenesis in Four Tree Legumes

2011 ◽  
Vol 2011 ◽  
pp. 1-8 ◽  
Author(s):  
Premananda Das
Keyword(s):  
Somatic Embryogenesis ◽  
Somatic Embryos ◽  
Zygotic Embryos ◽  
Dichlorophenoxyacetic Acid ◽  
Ms Medium ◽  
Embryogenic Calli ◽  
Friable Embryogenic Callus ◽  
Napthaleneacetic Acid ◽  
Acacia Catechu ◽  
2,4 Dichlorophenoxyacetic Acid

Somatic embryogenesis was achieved in four leguminous tree species, that is, Acacia catechu, Acacia arabica, Hardwickia binata, and Dalbergia sissoo using immature zygotic embryos as explants on Murashige and Skoog (MS) medium supplemented with 0.25–1.0 mg/l Kn (kinetin) and 2.0–3.0 mg/l 2,4-D (2,4-dichlorophenoxyacetic acid) or NAA (1-napthaleneacetic acid) and 3% sucrose. MS medium containing 2.0 mg/l 2,4-D and 1.0–1.5 mg/l Kn was noted to be most effective in inducing friable embryogenic callus (FEC); the number of somatic embryos per culture varied in MS medium supplemented with 1.0–2.0 mg/l 2,4-D or NAA and 0.25–1.5 mg/l kinetin. The maximum number of somatic embryos was obtained in MS medium containing 1.5–2.0 mg/l 2,4-D or NAA and 1.0–1.5 mg/l kinetin; proliferation of embryogenic calli was enhanced in cultures having 1.0–2.0 mg/l 2,4-D, 1.0–1.5 mg/l kinetin, and 400–600 mg/l L-Proline. The somatic embryos in various shapes and sizes after the first subculture on MS medium supplemented with 0.1 mg/l IAA and 0.25 mg/l BA; developed shoots and rooted in strength MS medium supplemented with 0.1 mg/l IBA or IAA. The somatic embryo-derived plantlets were transferred to the field after being hardened in the climate-controlled hardening chamber.

scholarly journals High Frequency of Somatic Embryogenesis and Recover of Fertile Cucumber Plants

HortScience ◽  
1990 ◽  
Vol 25 (7) ◽  
pp. 792-793 ◽  
Author(s):  
Paula P. Chee
Keyword(s):  
Somatic Embryogenesis ◽  
Growth Regulators ◽  
High Frequency ◽  
Somatic Embryos ◽  
Simple Procedure ◽  
Dichlorophenoxyacetic Acid ◽  
Ms Medium ◽  
Bipolar Structure ◽  
Hypocotyl Explants ◽  
2,4 Dichlorophenoxyacetic Acid

A simple procedure for regeneration of cucumber plants (Cucumis sativus L. cv. Poinsett 76) from cotyledon and hypocotyl explants has been developed. Somatic embryogenesis was induced on Murashige and Skoog (MS) salts and vitamins medium supplemented with 2,4-D at 2.0 mg·liter-1 and kinetin at 0.5 mg·liter-1. Development of embryos was accomplished on MS medium with NAA at 1.0 mg·liter-1 and kinetin at 0.5 mg·liter-1. Eighty-five percent of the mature somatic embryos formed showed a typical bipolar structure. All developed into morphologically normal plantlets when transferred to MS medium containing no growth regulators. Chemical name used: 2,4-dichlorophenoxyacetic acid (2,4-D).

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.

EFFICIENT CALLUS INDUCTION AND PLANT REGENERATION VIA SOMATIC EMBRYOGENESIS FROM IMMATURE LEAF-DERIVED PROTOPLASTS OF GROUNDNUT (ARACHIS HYPOGAEA L.)

1996 ◽  
Vol 44 (4) ◽  
pp. 387-396 ◽  
Author(s):  
Perumal Venkatachalam ◽  
Narayanasamypillai Jayabalan
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Arachis Hypogaea ◽  
Callus Cultures ◽  
Alternative Methods ◽  
Naphthalene Acetic Acid ◽  
Dichlorophenoxyacetic Acid ◽  
Ms Medium ◽  
High Yields ◽  
2,4 Dichlorophenoxyacetic Acid

High yields of protoplasts were obtained from immature leaves of aseptically grown plants of Arachis hypogaea using an enzyme solution containing cellulase 2.0% (w/v) and Macerozyme 1.0% (w/v) in 0.6 M mannitol. Isolated protoplasts were cultured in Kao's medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-benzylaminopurine (BAP). The protoplasts started to divide after 3–5 days of culture. Sustained divisions resulted in mass production of cell colonies and mini calli in 4 weeks. After 4 weeks, protoplast colonies were transferred to the Murashige and Skoog (MS) medium supplemented with a-naphthalene acetic acid (NAA) and BAP. Colonies proliferated into actively growing calli. Further attempts to regenerate plants from such calli were not successful. However, protoclones differentiated roots on the same medium. Alternative methods for plant regeneration from protoplast derived callus cultures were tried through somatic embryogenesis. Protoplast-derived calli treated with 2,4-D and BAP formed somatic embryos. Somatic embryogenesis began in the proembryo stage and proceeded from globular to dicotyledonary stage. Embryos were then transferred onto hormone-free MS medium for germination. Five to ten percent of these embryoids germinated and grew to plantlets. Regenerated plants were transferred to plastic cups and grown to maturity.

scholarly journals Somatic Embryogenesis in Carnation

HortScience ◽  
1992 ◽  
Vol 27 (1) ◽  
pp. 63-65 ◽  
Author(s):  
Les Frey ◽  
Yehoshua Saranga ◽  
Jules Janick
Keyword(s):  
Somatic Embryogenesis ◽  
Embryonic Development ◽  
Somatic Embryos ◽  
Single Cells ◽  
Dianthus Caryophyllus ◽  
Basal Medium ◽  
Dichlorophenoxyacetic Acid ◽  
Ex Vitro ◽  
Murashige And Skoog Medium ◽  
2,4 Dichlorophenoxyacetic Acid

Somatic embryogenesis was induced from internodal callus of `Scania', `Improved White Sim', and `Sandra' carnation (Dianthus caryophyllus L.). The optimum protocol for the induction of somatic embryogenesis included initiation of callus in liquid basal Murashige and Skoog medium supplemented with 3.0 μm 2,4-D followed by transfer to liquid basal medium lacking 2,4-D for embryo development. Somatic embryos originated from single cells and early embryonic development proceeded conventionally (i.e., via globular, heart-shaped, and torpedo stages), but clearly developed apical or root meristems were not always formed. A few embryos developed into seedlings and were acclimatized to ex vitro conditions. Chemical name used: 2,4-dichlorophenoxyacetic acid (2,4-D).

scholarly journals Development and regeneration of somatic embryos from leaves-derived calli of Coffea liberica

2020 ◽  
Vol 21 (12) ◽  
Author(s):  
FITRIA ARDIYANI ◽  
Edy Setiti Wida Utami ◽  
HERY PURNOBASUKI ◽  
SENJA APRILIA PARAMITA
Keyword(s):  
Somatic Embryogenesis ◽  
Embryo Development ◽  
Somatic Embryos ◽  
Economic Value ◽  
Genetic Material ◽  
Clonal Plants ◽  
Ms Medium ◽  
Cotyledonary Stage ◽  
Coffea Liberica

Abstract. Ardiyani F, Utami ESW, Purnobasuki H, Paramita SA. 2020. Development and regeneration of somatic embryos from leaves-derived calli of Coffea liberica. Biodiversitas 21: 5829-5834. Coffea liberica is an important and potentially commercial plant with a high economic value from the Coffea genus. Therefore, the availability of planting material is needed to increase productivity and ensure the sustainability of its farming. Somatic embryogenesis is a powerful propagation method used to produce clonal plants from limited genetic material. In the present research, we have shown that C. liberica could be successfully regenerated in vitro via somatic embryogenesis from leaves derived embryogenic callus. These calli were cultured on Murashige Skoog (MS) medium added with 1 mgL-1 BAP or in combination with 2.4 D (0.5, 1.0, 1.5 and 2 mgL-1) for embryo development induction. Furthermore, the medium containing only BAP was best for embryo development induction after culturing for 12 weeks, with the highest number of cotyledonary stage embryos (17.8%) and producing a total of embryo (20.2). Following cotyledonary stage embryo were cultured on new MS medium containing 0.5 mgL-1 BAP, 0.5 mgL-1 IAA, 0.5 mgL-1 NAA only, and 0.5 mgL-1 BAP in combination with 0.5 mgL-1 IAA or 0.5 mgL-1 NAA. Interestingly, the results showed that cotyledonary stage embryos were converted into complete plants at all treatment, but the MS medium containing 0.5 mgL-1 BAP was found to be the most effective in promoting regeneration with 2.6 leaves per-plantlet and height of 5.2 mm. Based morphological analysis confirm that the development of somatic embryo from leaves-derived calli of Coffea liberica started with the formation of embryo globular, heart, torpedo, cotyledonary stages, and finally conversion of cotyledonary embryo into complete plant.

scholarly journals 116 Somatic Embryogenesis and Organogenesis in Cowpeas

HortScience ◽  
1999 ◽  
Vol 34 (3) ◽  
pp. 461D-461 ◽  
Author(s):  
Lurline Marsh
Keyword(s):  
Somatic Embryogenesis ◽  
Indoleacetic Acid ◽  
Casein Hydrolysate ◽  
The Other ◽  
Dichlorophenoxyacetic Acid ◽  
Embryonic Axes ◽  
Embryo Formation ◽  
Stage Embryo ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Cotyledonary Stage

Four cowpea [Vigna unguiculata (L). Walp] genotypes; IT 82E-18, IT 82E-16, Pinkeye Purple Hull, and Coronet were tested for somatic embryo formation and embryogenesis. Explants were 3-week-old cotyledons from which the embryonic axes were removed. Cotyledons were cultured in eight media combinations representing modifications of two media, one containing Murashige and Skoog Basal salt with B5 vitamins (MSB), 500 mg/L casein-hydrolysate (CS), 500 mg/L sodium chloride, 3% sucrose, 0.7% agar, 2mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.5 mg/L benzylamino purine, and the other containing (MSB), 3% sucrose, 40 mg/L 2-4-D and 0.2% gellan gum. After 1 month, 40% to 100% of explants produced calli and few produced shoots. Subcultured shoots in MS with 0.1 mg/L indole-3-butyric acid (IBA) or with IBA and 0.5mg/L kinetin (KT) failed to produced roots. The only green cotyledonary stage embryo was produced on this latter medium. Subculture of calli in MSB containing CS, mannitol, sucrose, agar, indoleacetic acid, and KT produced cream-colored globular embryos, roots, and a few leaves.

scholarly journals CPPU IN THE MEDIUM FOR SEED GERMINATION PROMOTES EMBRYOGENESIS FROM SEEDLING EXPLANTS IN COMMON BEAN.

HortScience ◽  
1992 ◽  
Vol 27 (6) ◽  
pp. 617f-617
Author(s):  
Mohamed F. Mohamed ◽  
Paul E. Read ◽  
Dermot P. Coyne
Keyword(s):  
Somatic Embryogenesis ◽  
Seed Germination ◽  
Liquid Medium ◽  
Common Bean ◽  
Somatic Embryos ◽  
Casein Hydrolysate ◽  
Bipolar Structure ◽  
Seedling Explants ◽  
Explant Cultures ◽  
Cotyledonary Stage

Few studies on embryogenesis in common bean (Phaseolus vulgaris L.) have been reported and only the early stages of somatic embryogenesis were observed. Dry seeds from two common bean lines were germinated in darkness on L-6 medium containing 4% sucrose, 0.2 g casein hydrolysate /liter and 2.0 g phytagel /liter. The medium for seed germination was supplemented with 0, 2, 4 or 6μM forchlorfenuron (CPPU). Explants from cotyledonary leaves, petioles, hypocotyls and shoot apices were prepared from 14 day-old seedlings. Callus was derived from explant cultures incubated in darkness at 26C on the medium containing 4 μM 2,4-D and 1 μM Kinetin. The callus was transferred after 4 weeks into 125 ml Erlenmeyer flasks containing 50 ml liquid medium and placed on a gyrotary shaker (120 rpm) under cool-white light (12 μmol.m-2.s-1). The liquid medium was used with 2, 4 or 6 μM of 2,4-D alone or with zeatin supplements at relative concentrations of 0.25 and 0.5. Up to 200 somatic embryos from 40 to 50 mg callus inoculations were induced after 4 to 5 weeks. Callus derived from seedlings grown on CPPU-containing medium gave more repetitive somatic embryos. Cotyledonary stage embryos with clear bipolar structure were observed only from callus derived from seedlings grown on CPPU when transferred to suspension cultures containing 2,4-D and zeatin. All somatic embryos differentiated strong roots and some developed leaf-like structures on conversion medium.

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