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

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.

2016 ◽  
Vol 19 (2) ◽  
pp. 168
Author(s):  
Toni Herawan ◽  
Mohammad Na'iem ◽  
Sapto Indrioko ◽  
Ari Indrianto

Sandalwood (Santalum album L.) is native species of Indonesia, especially in East Nusa Tenggara, is oneof the twenty two species of the genus Santalum in the world. Sandalwood is an important tree because it hashigh economic value can produce sandal oil these can be used for perfumes, cosmetics, pharmaceuticals, andare often used in religious ceremonies. In vitro particularly somatic embryogenesis has been widely appliedin the propagation of sandalwood. The Objective of this research is to obtain regeneration of sandalwoodthrough somatic embryogenesis using leaves explant from various clones. Medium for embryo induction is MS(Murashige and Skoog, 1962) solid medium containing treatment of 2,4-D (2,4-Dichlorophenoxyacetic acid)at various concentrations. To the media 0,15 mg /l kinetin, 40 g/l sucrose, and 2,5 g/l gelrite were added.Culture were incubated in the dark. Medium for Embryo development (maturation) is MS solid mediumcontaining treatment of BAP (Benzyl-amino-purine) at various concentrations. To the media 0,01 mg /l NAA(Napthalene-acetic-acid), 40 g/l sucrose, and 2,5 g/l gelrite were added. Culture were incubated in the light. Tostudy the specifi c structure of sandalwood somatic embryo early detection was conducted using histologicalanalysis. Results of anova showed that the clones, media, and interaction between clones with media did notsignifi cantly affect the development of sandalwood callus percentage. Results of anova showed that the clonesand BAP concentration signifi cantly effect to the embryo development of sandalwood.


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.


1980 ◽  
Vol 58 (13) ◽  
pp. 1464-1473 ◽  
Author(s):  
Piyush Swami ◽  
V. Raghavan

The effects of 2,4-dichlorophenoxyacetic acid (2,4-D), gibberellic acid (GA), and abscisic acid (ABA) on the morphogenesis of gametophytes of Lygodium japonicum growing as longer-than-broad biplanar plates in red light and as broader-than-long biplanar plates in blue light were studied. Addition of 2,4-D or GA to the medium induced a change in the form of gametophytes from biplanar to filamentous in red light. Gametophytes growing in a medium containing 2,4-D in blue light were longer than broad, very much like gametophytes growing in the basal medium in red light. Although ABA generally retarded the growth of gametophytes in both light regimes, its presence in a medium containing 2,4-D nullified the effect of the latter, causing gametophytes to become plate-like in red light and short and stunted in blue light. Changes in the morphology of gametophytes induced by growth hormones were accompanied by corresponding changes in their length:width ratio and cell number.


2010 ◽  
Vol 70 (2) ◽  
pp. 361-366 ◽  
Author(s):  
MBB Cassanego ◽  
A Droste ◽  
PG Windisch

Regnellidium diphyllum is considered as endangered, occurring in the State of Rio Grande do Sul, Brazil, and a few adjoining localities in Uruguay, Argentina and the State of Santa Catarina. It grows in wetlands frequently altered for agricultural activities. Herbicides based on 2,4-dichlorophenoxyacetic acid (2,4-D) are widely used in these fields. The effects of 2,4-D on the germination of megaspores and initial sporophytic development of R. diphyllum were investigated. Six concentrations of 2,4-D (0.32; 0.64; 1.92; 4.80; 9.60 and 19.20 mg.L-1), and the control (0.00 mg.L-1), were tested in vitro, using Meyer's medium. Cultures were maintained in a growth chamber at 24 ± 1 °C, under artificial light with nominal irradiance of 110 µmol.m-2/s and 16 hours photoperiod. Megaspore germination was lower at 9.60 and 19.20 mg.L-1 of 2,4-D (56 and 48%, respectively), compared with the control (68%). Herbicide concentrations of up to 1.92 mg.L-1 did not significantly decrease the number of sporophytes formed. At 19.20 mg.L-1, no sporophytes were formed. The lengths of the primary root, primary and secondary leaves were greater at concentrations of 0.32 and 0.64 mg.L-1 of 2,4-D. Low concentrations of 2,4-D do not affect germination rates and initial development of R. diphyllum in a significant way. However, higher concentrations (9.60 and 19.20 mg.L-1) affect substantially the germination of the megaspores and interfere with the establishment of the species.


1989 ◽  
Vol 19 (2) ◽  
pp. 285-288 ◽  
Author(s):  
S. A. Merkle ◽  
A. T. Wiecko

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.


1996 ◽  
Vol 44 (4) ◽  
pp. 387-396 ◽  
Author(s):  
Perumal Venkatachalam ◽  
Narayanasamypillai Jayabalan

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.


2006 ◽  
Vol 131 (5) ◽  
pp. 586-592 ◽  
Author(s):  
Amílcar M.M. Duarte ◽  
Amparo García-Luis ◽  
Rosa Victoria Molina ◽  
Consuelo Monerri ◽  
Vicente Navarro ◽  
...  

A winter gibberellic acid (GA3) spray consistently reduced flower formation, but had a variable effect on the amount of first-grade fruit in the early harvest of `Clausellina' satsuma (Citrus unshiu Marc.), and in the long term these applications had no significant effect on the value of the crop. Auxin applications increased the amount of first grade-early harvested fruit, and increased crop value as compared to hand-thinned trees. No significant differences in yield or fruit grade could be found among the different auxin applications tried, namely an application of 20 mg·L-1 2,4-dichlorophenoxyacetic acid (2,4-D) at flowering, or applications of 25 mg·L-1 naphthaleneacetic acid (NAA), or 50 mg·L-1 2,4-dicholorophenoxypropionic acid (2,4-DP) at the end of fruitlet abscission. Apart from their effect on size, the auxin applications had only a marginal effect on fruit quality.


HortScience ◽  
2006 ◽  
Vol 41 (5) ◽  
pp. 1325-1329 ◽  
Author(s):  
Martín Mata-Rosas ◽  
Ángel Jiménez-Rodríguez ◽  
Victor M. Chávez-Avila

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.


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