Embryogenèse somatique et regénération de plantes à partir de protoplastes issus de suspensions cellulaires de céleri (Apium graveolens)

1991 ◽  
Vol 69 (7) ◽  
pp. 1583-1592
Author(s):  
C. Watin-de-Pontfarcy ◽  
C. Bigot
Keyword(s):  
Somatic Embryogenesis ◽  
Culture Cell ◽  
Cell Suspensions ◽  
Apium Graveolens ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Division Rate ◽  
Cell Division Rate ◽  
Variable Capacity ◽  
2,4 Dichlorophenoxyacetic Acid

Protoplasts were isolated from cell suspensions of Apium graveolens L. variety rapaceum (celeriac) initiated and subcultured on Gamborg medium (B5) with 2,4-dichlorophenoxyacetic acid (2.26 μM) and 6-benzylaminopurine (1.1 μM). The microcalli fraction above 450 μm was taken from 6-days-old cell suspensions and put into an enzyme mixture of cellulase Onozuka R-10 (2%) and pectolyase Y-23 (0.1%). Protoplasts were cultured on Lazar et al. modified medium with 2,4-dichlorophenoxyacetic acid (2.26 μM), naphthaleneacetic acid (0.89 μM), and zeatin (0.5 μM) at a density of 3 × 105∙mL−1. They were kept in darkness at alternating temperatures: 28 and 22 °C during 16 and 8 h, respectively. The culture medium was partially renewed after 7, 14, and 21 days of culture. Cell division rate represented about 30–40% of viable protoplasts. Addition of 2,4-dichlorophenoxyacetic acid (2.26 μM) in the microcalli plating medium improved embryogenic differentiation and subsequent plantlet isolation. Calli from protoplasts showed a variable capacity for somatic embryogenesis on Murashige and Skoog medium containing kinetin (1.4 μM). Plantlet growth was observed more particularly on microcalli plating medium without growth regulators. After acclimatization, a great number of plants with abnormal morphology was noted from calli cultivated on medium with kinetin. No variation of the normal ploidy level (2n = 2x = 22) has been recorded among the examined plants. Key words: Apium graveolens, protoplast, somatic embryogenesis, somaclonal variation, flowering.

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 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.

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 Somatic Embryogenesis and Plant Regeneration from Muscadine Grape Leaf Explants

HortScience ◽  
1993 ◽  
Vol 28 (1) ◽  
pp. 53-55 ◽  
Author(s):  
Carol Robacker
Keyword(s):  
Somatic Embryogenesis ◽  
Growth Regulators ◽  
Leaf Explants ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Vitis Rotundifolia ◽  
Regenerated Plants ◽  
Immature Leaf ◽  
Grape Leaf ◽  
2,4 Dichlorophenoxyacetic Acid

Immature leaf laminae and petioles of `Regale' and `Fry' muscadine grapes (Vitis rotundifolia Michx.) were cultured on Nitsch and Nitsch (NN) medium supplemented with 9.0 μm 2,4-D and 4.4 μm BA, and gelled with agar. Callus and original explant tissues were transferred to NN medium containing 10.7 μm NAA and 0.9 μm BA to proliferate embryogenic callus, which, when transferred to NN medium without growth regulators, yielded globular embryos. The embryos matured and germinated after being subcultured to fresh medium without growth regulators. Somatic embryogenesis incidence was greater from petioles than laminae: 90% of `Regale' and 50% of `Fry' petioles formed embryos, compared with 14% and 2% of laminae, respectively. Culturing germinated somatic embryos on NN medium with 1 μm BA enhanced shoot growth. Regenerated plants flowered and appeared morphologically normal. Chemical names used: N-(phenylmethyl)- 1H -purin-6-amine (BA); 2,4-dichlorophenoxyacetic acid (2,4-D); α- naphthaleneacetic acid (NAA).

scholarly journals Impact of Osmotica and Plant Growth Regulators on Somatic Embryogenesis of Date Palm

2019 ◽  
Vol 7 (3) ◽  
pp. 296-303
Author(s):  
Mouaad Amine Mazri ◽  
Ilham Belkoura ◽  
Reda Meziani ◽  
Hajar Es-Saoudy ◽  
Fahd Rachad ◽  
...  
Keyword(s):  
Somatic Embryogenesis ◽  
Date Palm ◽  
Dichlorophenoxyacetic Acid ◽  
Adventitious Bud ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Mature Embryos ◽  
Maturation Medium ◽  
Semi Solid ◽  
2,4 Dichlorophenoxyacetic Acid

An efficient somatic embryogenesis system is reported for date palm cv. Al-Fayda, a genotype resistant to the bayoud disease. Callus induction was achieved from adventitious bud explants cultured for 6 months on semi-solid Murashige and Skoog (MS) medium containing 4.5 μM 6-(dimethylallylamino) purine (2iP) and various concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) or picloram. The highest somatic embryogenesis frequency (89%) was obtained on MS medium supplemented with 225 μM 2,4-D. Subsequently, embryogenic cultures were transferred to agitated liquid MS medium (maturation medium) containing various concentrations of mannitol, polyethylene glycol (PEG) or sorbitol. The highest rate of somatic embryo maturation (71.4 mature embryos per 100 mg callus) was achieved on the medium supplemented with 40 g l-1 PEG. Mature somatic embryos were then transferred to MS medium supplemented with gibberellic acid (GA3) or 1-naphthaleneacetic acid (NAA) and 6-benzylaminopurine (BAP) at various concentrations. The highest frequency of germination and conversion (26%) was obtained on the medium containing 5 μM NAA and 5 μM BAP. The developed plants were then transferred to ex vitro conditions, where a survival rate of 77.02% was observed. The regeneration protocol established in the present investigation will be used for mass propagation of date palm cv. Al-Fayda.

scholarly journals EMBRIOGENESIS SOMATIK IN VITRO DAN REGENERASI PLANLET DARI TIGA VARIETAS ALFALFA (Medicago sativa L.)

2019 ◽  
Vol 6 (1) ◽  
pp. 83
Author(s):  
Sulastri Sulastri ◽  
Winda Nawfetrias ◽  
Djatmiko Pinardi ◽  
Henti Rosdayanti
Keyword(s):  
Somatic Embryogenesis ◽  
Medicago Sativa ◽  
Callus Induction ◽  
Plantlet Regeneration ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Medicago Sativa L ◽  
Embryogenic Callus Induction ◽  
2,4 Dichlorophenoxyacetic Acid

In Vitro Somatic Embryogenesis and Plantlet Regeneration of Three Varieties of Alfalfa (Medicago sativa L.)ABSTRACTAlfalfa (Medicago sativa L.) is a valuable plant as a source of food for animal, forage, pharmaceutical, medicine, food supplement, and human consumption.  In vitro selection technology combined with induction or spontaneous mutagenesis has been effective in altering or isolating genetic variability for desirable characters.  Consequently, a reproducible in vitro propagation technique of that plant is mandatory. The aim of the research was to obtain information on the embryogenic callus induction, somatic embryogenesis, and plantlet regeneration of three varieties of alfalfa. The results showed that an optimum embryogenic callus induction (82%) was obtained on Murashige & Skoog (MS) basal medium containing 2 ppm 2,4-dichlorophenoxyacetic acid (2,4-D), 2 ppm kinetin and 2 ppm a-naphthaleneacetic acid (NAA). Those embryogenic calli could subsequently develop into somatic embryos, which germinated and regenerated into normal plantlets on R1 medium consisting of MS nutrients without the addition of plant growth regulator.Keywords: alfalfa, callus, embryogenic, plantlets, regeneration ABSTRAKAlfalfa (Medicago sativa) adalah tanaman berharga sebagai sumber makanan untuk hewan, yaitu hijauan pakan ternak, farmasi, obat-obatan, suplemen makanan dan konsumsi manusia. Teknologi seleksi in vitro yang dikombinasikan dengan induksi atau mutagenesis spontan telah terbukti efektif dalam mengubah atau mengisolasi variabilitas genetik untuk karakter yang diinginkan. Oleh sebab itu, keberhasilan teknik perbanyakan in vitro yang telah terbukti dapat direproduksi dari tanaman tersebut menjadi syarat yang harus terpenuhi. Tujuan dari penelitian ini adalah untuk mendapatkan informasi mengenai induksi kalus embriogenik, embriogenesis somatik dan regenerasi planlet dari tiga varietas alfalfa. Hasil penelitian menunjukkan bahwa induksi kalus embriogenik optimal (82%) didapat pada media Murashige & Skoog (MS) dengan  2 ppm 2,4-dichlorophenoxyacetic acid (2,4-D), 2 ppm kinetin dan 2 ppm a-naphthaleneacetic acid (NAA). Kalus embriogenik tersebut dapat membentuk embrio somatik, embrionya berkecambah dan beregenerasi membentuk planlet normal pada perlakuan media R1 yaitu nutrisi MS tanpa penambahan zat pengatur tumbuh.Kata Kunci: alfalfa, embriogenik, kalus, planlet, regenerasi

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.

Initiation and morphological development of somatic embryoids from barley cell cultures

1984 ◽  
Vol 62 (6) ◽  
pp. 1245-1249 ◽  
Author(s):  
L. S. Kott ◽  
K. J. Kasha
Keyword(s):  
Somatic Embryogenesis ◽  
Abscisic Acid ◽  
Gibberellic Acid ◽  
Cell Cultures ◽  
Dichlorophenoxyacetic Acid ◽  
Morphological Development ◽  
Low Concentrations ◽  
The Media ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Barley Cell

Somatic embryogenesis was induced in callus previously initiated from immature embryos of barley. These cultures ranged in age from 6 weeks to 30 months. Embryoids were readily initiated from homogenized suspension-grown aggregates when plated on modified B5 media with 2,4-dichlorophenoxyacetic acid. Low concentrations (0.1 and 0.05 mg∙L−1) of abscisic acid promoted further maturation of embryoids, while gibberellic acid (1 mg∙L−1) and kinetin (0.1 mg∙L−1) were used in the media to encourage embryoid germination. The development of somatic embryoids from initiation through maturation and germination is described.

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.'

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