Embryolike structures from cotyledons and ripe embryos of Norway spruce (Piceaabies)

1986 ◽  
Vol 16 (3) ◽  
pp. 664-668 ◽  
Author(s):  
P. Krogstrup

Embryos from imbibed ripe seeds and cotyledon expiants of 7-day-old Norway spruce (Piceaabies (L.) Karst.) seedlings produced the early stages of somatic embryogenesis. Using a modified Murashige and Skoog medium, a whitish, glossy callus was induced consisting of translucent cells embedded in a mucilaginous cloudy matrix. This embryogenic callus formed on the surface of explants treated first with N-6-benzyladenine followed by 2,4-dichlorophenoxyacetic acid + 6-furfurylaminopurine (kinetin) + N-6-benzyladenine. Transfer of this callus to media lacking growth regulators resulted in the formation of numerous bipolar embryoids with suspensorlike structures. These embryoids strongly resembled repressed embryos in polyembryonic seeds.

HortScience ◽  
1992 ◽  
Vol 27 (1) ◽  
pp. 63-65 ◽  
Author(s):  
Les Frey ◽  
Yehoshua Saranga ◽  
Jules Janick

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).


2015 ◽  
Vol 16 (1) ◽  
pp. 17
Author(s):  
Wardatus Sholeha ◽  
Bambang Sugiharto ◽  
Dwi Setyati ◽  
Parawita Dewanti

Induction of somatic embryogenesis in sugarcane requires the composition Plant Growth Hormone (PGH) appropriate. Utilizing of PGH (2,4-D and kinetin) is expected to induce sugarcane somatic embryogenesis. The purpose of this study was to obtain the concentration of 2,4-D and kinetin that effective for the multiplication of sugarcane var. NXI 1-3 through somatic embryogenesis. This study consists of four stages: callus induction, callus proliferation, regeneration of shoots and encapsulation. The plant material used is a spindle leaf sugarcane var. NXI 1-3. Callus induction used 2,4-D with different concentration (2 ppm, 3 ppm and 4 ppm). Callus proliferation used 2,4-D with concentration 1 ppm and 2 ppm. Regeneration of shoots used kinetin 0,5 ppm. The results are showed that the optimal induction of embryogenic callus on medium MS + sucrose 30 g / L + CH 300 ppm + 300 ppm PVP + 2,4-D 4 ppm as indicated by the high percentage of explants forming embryogenic callus that is equal to 40% and the respective concentration 2 ppm and 3 ppm is 33,3% and 37,5%. In proliferation stage, the development callus optimal on medium MS + sucrose 30 g / L + CH 300pm + PVP 300 ppm + 2,4-D 2 ppm and formulations for regeneration shoot on medium MS + sucrose 30 g / L + kinetin 0.5 ppm. The result of encapsulation can be shaped 100 sythetic seed. Keywords: Somatic embryogenesis, spindle leaf, kinetin, 2,4-D


CORD ◽  
2016 ◽  
Vol 32 (2) ◽  
pp. 6
Author(s):  
Carlos Oropeza

This paper focuses on the research efforts carried out by CICY in Mexico on micropropagation of coconut. They started during the nineties in collaboration with Wye College (UK) and ORSTOM-CIRAD (France), with the development of a protocol that was reproducible and more efficient than previous ones, based on plumule explants grown in different media based on Y3 medium added with activated charcoal, gelling agent and of particular importance growth regulators 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-benzylaminopurine (BAP). Within the next decade basic research was carried out to study the process of somatic embryogenesis from plumule explants, with an approach including morpho-histological, physiological, biochemical and molecular points of view, in order to gain knowledge that could be useful to further improvement of the process. Also different practical approaches were tested including changes in the media formulation, embryogenic callus multiplication and secondary somatic embryogenesis. As a result a highly efficient protocol was developed that could potentially yield over a hundred thousand somatic embryos from a single plumule explant. Embryos were able to germinate and convert to plantlets, that after planting, successfully grew to sexual maturity and fruit production. This protocol is currently being scaled up to a semi-commercial level. Also within the past five years, a protocol using rachilla explants has been developed for the production of embryogenic callus and its multiplication, and embryos produced were able to germinate and convert to plantlets, setting the basis to develop a process for massive  propagation of coconut, such as the one already developed using plumule explants.


2014 ◽  
Vol 51 (2) ◽  
pp. 173-178 ◽  
Author(s):  
Aurelia Ślusarkiewicz-Jarzina ◽  
Maciej Zenkteler ◽  
Barbara Podlewska

Plants were obtained from cultured in vitro leaves of Chrysanthemum morifolium Ram. cv. Bronze Bornholm. The leaves were inoculated on Murashige and Skoog medium (MS) supplemented with cytokinins (kinetin - KIN, zeatin - ZEA, 6-benzyloaminopurine - BAP) and auxins (2,4-dichlorophenoxyacetic acid - 2,4-D, α-naphtaleneacetic acid - NAA, 3-indolilacetic acid - IAA, p-fluorophenylalanine - PFA) in various combinations and concentra-tions. The most suitable medium was that one which contained 4 mg/l KIN, 2 mg/l NAA and 50 mg/l PFA.


HortScience ◽  
1990 ◽  
Vol 25 (7) ◽  
pp. 792-793 ◽  
Author(s):  
Paula P. Chee

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).


1988 ◽  
Vol 18 (7) ◽  
pp. 937-941 ◽  
Author(s):  
M. Rafique Uddin ◽  
Martin M. Meyer Jr. ◽  
J. J. Jokela

Plantlets were obtained by organogenesis from cultured anthers of Populusdeltoides (Bartr.). Anthers formed callus in the dark on modified Murashige and Skoog medium supplemented with 9.0 μM 2,4-dichlorophenoxyacetic acid and 4.7 μM kinetin. Anther calli were differentiated into shoots by sequential transfer in the light onto Murashige and Skoog medium containing 4.4 μM benzylamino purine and 1.1 μM naphthaleneacetic acid for 4 weeks, followed by several transfers to woody plant medium with 2.2 μM benzylamino purine and 1.1 μM naphthaleneacetic acid. The shoots that formed were rooted by excising and transferring to woody plant medium supplemented with 1.0 μM indole-3-butyric acid. A few of these plants were found to be haploid. Two plants developed male terminal inflorescences, but died shortly thereafter.


HortScience ◽  
2017 ◽  
Vol 52 (1) ◽  
pp. 142-145 ◽  
Author(s):  
Modeste Kan Kouassi ◽  
Jane Kahia ◽  
Christophe N’guessan Kouame ◽  
Mathias Gnion Tahi ◽  
Edmond Kouablan Koffi

The effect of plant growth regulators on callus and somatic embryogenesis induction in four Cocoa (Theobroma cacao) genotypes was studied. Flower explants were harvested early in the morning and cultured on Driver and Kuniyuki Walnut (DKW) medium supplemented with 1 mg·L−1 of five auxins type (2,4 dichlorophenoxyacetic acid (2,4-D), 3,4 dichlorophenoxyacetic acid (3,4-D), 2,4,5 trichlorophenoxyacetic acid (2,4,5-T), 4-amino-3,5,6-trichloropicolinic acid (picloram), and 3,6-dichloro-2-methoxybenzoic acid (dicamba) in combination with 0.25 or 0.5 mg·L−1 of two cytokinins type (benzylaminopurine (BAP) and 6-furfurylaminopurine [kinetin (Kin)] in a factorial experiment. The plant growth regulators 2,4-D and 2,4,5-T proved to have a broad spectrum action on somatic embryogenesis induction compared with 3,4-D or picloram. There were no significant differences between the two concentrations of cytokinins. However, Kin was found to be more effective in promoting somatic embryogenesis than BAP. Combining 1 mg·L−1 2,4,5-T or 2,4-D with 0.25 mg·L−1 Kin had a broad spectrum action on embryogenesis induction. On the other hand, combining mg·L−1 picloram with 0.5 mg·L−1 Kin or 1 mg·L−1 3,4-D with 0.25 mg·L−1 Kin was only able to induce somatic embryogenesis in a few of the genotypes evaluated. The protocol developed during the current study differs from earleir works as the callus (derived from explants cultured on DKW media) was taken directly to embryo development media as opposed to earlier works in which the callus was taken through a secondary media before being transferred to an embryo development media.


1986 ◽  
Vol 16 (2) ◽  
pp. 420-422 ◽  
Author(s):  
S. A. Merkle ◽  
H. E. Sommer

Tissue cultures of yellow poplar (Liriodendrontulipifera L.) were initiated from immature and mature zygotic embryos. Nodular embryogenic callus developed from a low percentage of the cultures initiated from immature embryos on solid media supplemented with 2,4-dichlorophenoxyacetic acid, 6-benzyladenine, and casein hydrolysate. Embryoids differentiated from these culture lines within 1 month following transfer of embryogenic callus to hormone-free solid media. Although most embryoids appeared abnormal, embryoids with well-formed cotyledons and radicles were capable of developing into normal plantlets.


1993 ◽  
Vol 23 (5) ◽  
pp. 835-838 ◽  
Author(s):  
Paula M. Pijut

Immature cotyledonary explants excised from developing fruits of Juglanscinerea L. were cultured in vitro to induce regeneration of somatic embryos. Somatic embryos were initiated directly on cotyledons collected 9 weeks postanthesis and cultured on a Driver and Kuniyuki medium supplemented with 250 mg/L L-glutamine, 0.01 mg/L indole-3-butyric acid, 1 mg/L 6-benzylaminopurine, and 2 mg/L kinetin for 3 weeks, prior to transfer to hormone-free Driverand Kuniyuki medium. Embryogenic callus was initiated on explants collected 8–11 weeks postanthesis and cultured on two different media formulations containing 0.25 mg/L 6-benzylaminopurine and 2 mg/L 2,4-dichlorophenoxyacetic acid for 3 weeks, prior to transfer to hormone-free media. Globular to mature somatic embryos were differentiated, and conversion of somatic embryos into whole plants was incomplete. Competence of embryogenic callus was maintained for 1 year with regular subculturing on hormone-free media.


Author(s):  
Ketut Anom Wijaya ◽  
Adi Prawoto ◽  
Syrril Ihromi

Cocoa (Theobroma cacao L.) like most tropical trees is recalcitrant in tissue culture. Somatic embryogenesis is generally efficient micropropagation technique to multiply elite material. However, Somatic embryogenesis in cocoa is difficult and this species is considered as recalcitrant. One of the factors often considered as a component of in vitro recalsitrance is a high phenolic content and oxidation of these compounds. In cocoa tissue culture accumulate large amounts of poliphenolics compounds which probably impair further development. This study was conducted to investigate the composition of phenolic compounds in cocoa flower and leaves, and their changes troughout the somatic embryogenesis process. Calli were induced in cacao floral and leaves explants on a half-strenght Murashige and Skoog medium containing 30 g/L Glucose and combination of 2,4 dichlorophenoxyacetic acid (2,4 D) with kinetin (kin). Total polyphenol content was observed on Sulawesi 1 cocoa clone. Embryogenic and non-embryogenic callus were also compared. The percentage of callus production from flower tissue is 85%, percentage of embryogenic callus 40 %, although  the percentage of somatic embryo production from embryogenic callus callus is 70%. The conservation of callus into somatic embryos followed by decline in phenol content and an increase in peroxidase. The synthesis kinetics for these compounds in calli, under different somatic embryogenesis conditions, revealed a higher concentration under non-embryogenic conditions. So that, phenolic compound can influence the production of calli and an absence the phenolic compound can enhance production of somatic embryo.Kata kunci: Theobroma cacao L., polifenol, embrio somatik, kalus, flavonoid, katekin, in vitro recalcitance


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