Somatic embryogenesis and plant regeneration from cell suspension derived protoplasts of Solanum melongena (eggplant)

1986 ◽  
Vol 64 (2) ◽  
pp. 355-361 ◽  
Author(s):  
S. Gleddie ◽  
W. A. Keller ◽  
G. Setterfield
Keyword(s):  
Somatic Embryogenesis ◽  
Cell Suspension ◽  
Somatic Embryos ◽  
Solanum Melongena ◽  
Cell Suspension Cultures ◽  
Callus Cultures ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Subsequent Regeneration

Cell suspension cultures of eggplant (Solanum melongena L.) were initiated from embryogenic callus cultures and maintained in medium supplemented with either 1-naphthaleneacetic acid (NAA) or 2,4-dichlorophenoxyacetic acid (2,4-D). Higher yields of protoplasts were obtained from cells grown in 2,4-D than in NAA. The efficiency of cell division was also greater in protoplast cultures derived from cells grown in the presence of 2,4-D. Protoplast-derived cells formed somatic embryos in modified Kao or Nagata and Takebe media which were supplemented with 1 mg/L 2,4-D. Early stages of embryogenesis were observed in liquid medium; however, these embryos and associated cell colonies were transferred onto agar-solidified medium for subsequent regeneration. Mature plants were established in soil in the greenhouse.

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 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.

scholarly journals Enhancement of Somatic Embryogenesis and Production of Developmentally Arrested Embryos in Nigella sativa L.

HortScience ◽  
2004 ◽  
Vol 39 (2) ◽  
pp. 321-323 ◽  
Author(s):  
Hamid Elhag ◽  
Mahmoud M. El-Olemy ◽  
Mansour S. Al-Said
Keyword(s):  
Somatic Embryogenesis ◽  
Liquid Medium ◽  
Somatic Embryos ◽  
Nigella Sativa ◽  
Basal Medium ◽  
Callus Cultures ◽  
Naphthaleneacetic Acid ◽  
Root Explants ◽  
Friable Callus ◽  
2,4 Dichlorophenoxyacetic Acid

Somatic embryogenesis of Nigella sativa was investigated with the objective of inducing and isolating somatic embryos for biosynthetic studies. Callus cultures were initiated from leaf, stem, and root explants of axenic seedlings on MSB5 basal medium supplemented with kinetin (0.46 μm) and 2,4-D (4.5 or 13.5 μm) or NAA (5.4 or 16.2 μm) in the dark. Cultures initiated and subcultured on medium containing NAA produced friable callus with numerous roots regardless of explant type. Cultures initiated, subcultured, or both, on medium with low 2,4-D concentration produced shiny embryogenic masses. These cultures differentiated into somatic embryos on medium containing NAA. The embryos developed into leafy structures on basal medium devoid of growth regulators. When the embryogenic callus was transferred to liquid medium containing NAA, numerous embryos and clusters of embryos were released into the liquid medium but, in contrast to solid medium, development remained arrested at the early embryonic stages. The developmentally arrested embryos were tested for production of active constituents of N. sativa oil. Chemical names used: 2,4-dichlorophenoxyacetic acid (2,4-D); α-naphthaleneacetic acid (NAA); kinetin (K).

In vitro culture of Bouvardia ternifolia

1982 ◽  
Vol 60 (6) ◽  
pp. 917-921 ◽  
Author(s):  
Leonor Fernandez ◽  
Estela Sanchez de Jimenez
Keyword(s):  
Indoleacetic Acid ◽  
Cell Suspension Cultures ◽  
Callus Cultures ◽  
Suspension Cultures ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Napthaleneacetic Acid ◽  
Leaf Tissues ◽  
2,4 Dichlorophenoxyacetic Acid

Callus cultures were induced from radicle and leaf tissues of Bouvardia ternifolia (trompetilla). Optimum growth regulator concentrations for radicle callus cultures were 1 mg/L 2,4-dichlorophenoxyacetic acid and 0.005 mg/L kinetin; for leaf callus they were either 2 mg/L naphthaleneacetic acid and 0.002 mg/L benzylaminopurine or 5 mg/L of idoleacetic acid and 0.01 mg/L kinetin. Callus has been maintained in culture for nearly 3 years with a very rapid growth rate.A generation time of approximately 24 to 28 h was obtained for batch cell suspension cultures. Production of protoplasts from suspension cultures was optimized with a yield of 70 to 90%. Protoplast culture was achieved in droplets of fresh medium with 2 mg/L napthaleneacetic acid, 0.01 mg/L benzylaminopurine, and0.5 M mannitol. After 2 years, callus in culture still retained its organogenetic capacity. An average of 18 complete plantlets from approximately 2 g of callus can be obtained after transfer to medium with 0.1 mg/L indoleacetic acid and 0.1 mg/L benzylaminopurine.

scholarly journals Effects of Plant Growth Regulators and Sugars on Ehretia asperula Zoll. et Mor. Cell Cultures

2021 ◽  
Author(s):  
P.T.M. Tram ◽  
N.K. Suong ◽  
L.T.T. Tien
Keyword(s):  
Cell Suspension ◽  
Callus Induction ◽  
Malignant Tumors ◽  
Single Cells ◽  
Cell Suspension Cultures ◽  
Suspension Cultures ◽  
Dichlorophenoxyacetic Acid ◽  
Human Immune System ◽  
Friable Callus ◽  
2,4 Dichlorophenoxyacetic Acid

Background: Belonging to the Boraginacae family, Ehretia asperula Zoll. et Mor., called “Xa den”, is a precious medicinal plant also known as the “cancer tree” by the Muong ethnic group in Hoa Binh Province, Vietnam. Xa den has been demonstrated to inhibit the development of malignant tumors, reduce oxidation and enhance the human immune system. This research focused on examining friable callus induction from young stems of Ehretia asperula Zoll. et Mor. Methods: Samples of Xa den were less than two years old. Young stems with 2 to 6 leaves served as explants for callus induction. Explants placed on autoclaved B5 nutrients incubated at 25oC, in the dark. The testing factors were concentrations of 2,4-Dichlorophenoxyacetic acid (2,4-D) and Benzyl adenine (BA), types and concentrations of sugars.Result: Friable callus was induced on B5 medium with 0.4 mg/L of 2,4-D, 0.1 mg/L of BA and 30 g/L of glucose at the highest rate (100%). Additionally, callus grew best after 5 weeks of culture weighing 0.194 g. Friable callus was used as material for cell suspension cultures. After two weeks, the Xa den cell suspension cultures contained single cells and small cell clumps. The liquid medium had changed from dark yellow to light brown.

scholarly journals Establishment of Morphogenic and Regeneration Ability in two Indica Rice Cell Suspension Cultures after Exposure to NaCl

1970 ◽  
Vol 19 (2) ◽  
pp. 185-197
Author(s):  
T.L. Aditya
Keyword(s):  
Cell Suspension ◽  
Callus Induction ◽  
Somatic Embryos ◽  
Indica Rice ◽  
Cell Suspension Cultures ◽  
Nacl Stress ◽  
Callus Cultures ◽  
Regeneration Ability ◽  
Rice Varieties

An efficient protocol was developed for in vitro morphogenic ability along with plantlet regeneration of two Bangladeshi indica rice varieties (BR24 and BR26) via somatic embryogenesis by applying 50 mM NaCl stress in callus induction and suspension initiation media. Osmotic stress was induced by NaCl (50, 100, 150, 200 and 250 mM) on the cell growth in suspension maintenance media. In viability test stress adapted cells showed 85 - 95% viability up to 200 mM NaCl compared with stress shocked (MS1-50) and control (MS1-0) treatments. Higher stress adapted cells showed growth retardation and the induction of plasmolysis. For both genotypes somatic embryos were obtained in both MS based liquid and semisolid media with or without 50 and 100 mM NaCl. Cell suspension-derived micro-calli were partially desiccated (6 - 12 hr) and subsequently maintained in MS1 callus induction media supplemented with proline (12 mM), ABA (2 mg/l) and 0.6% phytagel in the presence or absence of 50 and 100 mM NaCl. Subsequently, desiccated somatic embryos were transferred in MS based regeneration media with or without 50 and 100 mM NaCl. Proline mediated callus was found to be more effective in embryo differentiation than ABA. Partial desiccation dramatically enhanced callus growth and partially increased regeneration percentage. BR24 showed a better regeneration response producing plantlets in presence of proline in control media while BR26 restored regeneration potential in the presence of ABA and 100 mM NaCl. Plantlets regenerated from salt stressed callus cultures were then grown in compost in a glasshouse and produced normal, fertile plants.  Key words: Indica rice, Cell suspension, Morphogenic, Regeneration D.O.I. 10.3329/ptcb.v19i2.5436 Plant Tissue Cult. & Biotech. 19(2): 185-197, 2009 (December)

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.

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.

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