scholarly journals Somatic Embryogenesis and Massive Shoot Regeneration from Immature Embryo Explants of Tef

2011 ◽  
Vol 2011 ◽  
pp. 1-7 ◽  
Author(s):  
Likyelesh Gugsa ◽  
Jochen Kumlehn

Tef (Eragrostis tef) provides a major source of human nutrition in the Horn of Africa, but biotechnology has had little impact on its improvement to date. Here, we report the elaboration of an in vitro regeneration protocol, based on the use of immature zygotic embryos as explant. Explant size was an important determinant of in vitro regeneration efficiency, as was the formulation of the culture medium. Optimal results were obtained by culturing 0.2–0.35 mm embryo explants on a medium containing KBP minerals, 9.2–13.8 μM 2,4-dichlorophenoxyacetic acid, 6 mM glutamine, and 0.5% Phytagel. Although this protocol was effective for both the improved cultivar “DZ-01-196” and the landrace “Fesho”, the former produced consistently more embryogenic tissue and a higher number of regenerants. An average of more than 2,800 shoots could be obtained from each “DZ-01-196” explant after 12 weeks of in vitro culture. These shoots readily formed roots, and plantlets transferred to soil were able to develop into morphologically normal, fertile plants. This regeneration and multiplication system should allow for the application of a range of biotechnological methods to tef.

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.


2017 ◽  
Vol 65 (1) ◽  
pp. 80 ◽  
Author(s):  
Bilan Huang ◽  
Li Xu ◽  
Kelie Li ◽  
Yunlu Fu ◽  
Zhiying Li

An in vitro protocol for Callerya speciosa (Champ.) Schot regeneration through embryogenesis was developed using the anthers as the explants. The late uninucleate stage of the microspore was optimal for the anther culture of C. speciosa. Embryonic callus was induced on a MS basal medium supplemented with 4.4 µM 6-benzylaminopurine (BA) and 9.04 µM 2,4-dichlorophenoxyacetic acid (2,4-D). Embryos were obtained on MS medium supplemented with 2.2 µM BA and 0.5 µM naphthaleneacetic acid (NAA). The highest percentage (16.7%) of embryos was achieved using the culture medium MS + 0.25 µM NAA + 1.1 µM BA. The highest percentage of embryos that developed into plants was 18.3%. However, haploid plants were not observed, which may have been due to the collection of the calli from the anther wall. The results presented here demonstrate the establishment of a highly efficient and rapid system for regenerating C. speciosa using anther cultures.


2014 ◽  
Vol 14 (2) ◽  
pp. 108-115 ◽  
Author(s):  
Sandra Zorat Cordeiro ◽  
Naomi Kato Simas ◽  
Anaize Borges Henriques ◽  
Alice Sato

Mandevilla guanabarica is an endemic plant from Brazil, with pharmacological and ornamental potential, both unexplored. This study established the best culture medium for in vitro plant maintenance, efficient protocol for its regeneration, and callogenesis from different explants excised from in vitro-grown plants. Woody plant medium with double boron concentration (WPM B) plus 2.27 µM thidiazuron or 0.49 µM 2-isopentenyladenine provided multiplication rates higher than 1:6. Shoots were 100% rooted on WPM B. After acclimatization, plants showed 83% survival. For callogenesis, the use of MS media supplemented with high concentrations of picloram or 2,4-dichlorophenoxyacetic acid produced, respectively, friable or compact non-morphogenic calluses from different types of explants. This micropropagation protocol allows the production of seedlings of M. guanabarica for ornamental or commercial uses, and for conservation purposes; calluses can be used to establish suspension cultures in prospecting for bioactive compounds.


Plants ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 761
Author(s):  
Arun Kumar Khajuria ◽  
Christophe Hano ◽  
Narendra Singh Bisht

Viola canescens Wall. ex. Roxb. is an important but threatened medicinal herb found at 1500–2400 m above mean sea level in the Himalayas. Overexploitation and habitat preference have put the plant under serious threat. Thus, the present study was undertaken to develop an efficient protocol for in vitro propagation via somatic embryogenesis. The results revealed that plant can be regenerated successfully through somatic embryogenesis using leaf derived calli. Regular subculturing of calli on Murashige and Skoog (MS) medium with 2,4-dichlorophenoxyacetic acid (2,4-D)/indole-3-butyric acid (IBA)/kinetin (Kn) and varying combinations of 2,4-D+Kn induced somatic embryogenesis. The maximum average number of somatic embryos (SE) (19.15 ± 2.66) was induced on the medium with 0.15 + 0.05 mg L−1 of 2,4-D and Kn, respectively, and this medium was used as a control. To enhance somatic embryo induction, the control MS medium was supplemented with l-glutamine (200–400 mg L−1) and casein hydrolysate (1–4%). The maximum average number of SE (27.66 ± 2.67) and average mature SE (13.16 ± 3.48) were recorded on the medium having 2 % l-glutamine and 50 mg L−1 casein hydrolysate. The induced SE were asynchronous, so, to foster their maturation, the culture medium (free from growth regulators) was supplemented with abscisic acid (ABA) and silver nitrate (AgNO3). The maximum average number (35.96 ± 3.68) of mature SE was noticed on MS medium supplemented with 1.5 mg L−1 ABA. Mature embryos had two well-developed cotyledons and an elongated hypocotyl root axis. The development of SE into plantlets was significant for embryos matured on the medium with AgNO3 and ABA, with 86.67% and 83.33% conversion on the medium with 0.20 mg L−1 6-benzylaminopurine (BAP). The plantlets thus produced acclimatized in a growth chamber before being transferred to the field, which showed 89.89% survival. The plants were morphologically similar to the mother plant with successful flowering.


Author(s):  
Kicia K. P. Gomes- Copeland ◽  
Izulmé R. I. Santos ◽  
Amanda G. Torres ◽  
João V. D. Gomes ◽  
Fabrício T. C. de Almeida ◽  
...  

Amaryllidaceae include plant species that present alkaloids with analgesic, anti-cancer, anti-bacterial, anti-viral, anti-fungal and anti-malarial activities. Due to this pharmacological value, several species of this family have been widely studied and among them is White lilly, Crinum americanum. The objective of this work was to induce callogenesis on leaf explants of C. americanum cultivated in vitro for future production of alkaloids. Leaf explants were grown on a culture medium (solid) Murashige and Skoog (1962) supplemented with different concentrations and combinations of plant growth regulators, auxin 2,4-dichlorophenoxyacetic acid and cytokinin 6-benzylaminopurine and their effect on callogenesis assessed for percentage oxidation and explants responsive to callus induction. Callus formation started 10 days after hormone inoculation, and within 30 days after inoculation the best callogenesis and callus biomass growth were observed in medium containing 2.5 mg L-1 of 2,4-dichlorophenoxyacetic acid and 10 mg L-1 of 6-benzylaminopurine. The lowest percentage of oxidation was observed on explants cultivated on medium containing 5 mg L-1 of 6-benzylaminopurine and 2.5 mg L-1 of 2,4-dichlorophenoxyacetic acid. The calli obtained were compact and embryogenic. This work contributes not only to future studies on in vitro callogenesis of this species, but also to a possible protocol for the production of alkaloids of interest from cell suspension cultures produced in vitro. This is the first report of callus formation in Crinum americanum explants.


2007 ◽  
Vol 59 (3) ◽  
pp. 199-202 ◽  
Author(s):  
Dragana Stojicic ◽  
Branka Uzelac ◽  
Dusica Janosevic ◽  
Ljubinka Culafic ◽  
Snezana Budimir

The potential for somatic embryogenesis in zygotic embryo and megagametophyte cultures of Pinus heldreichii was examined. Somatic embryogenesis was initiated from megagametophytes containing immature zygotic embryos at early stages of development. An induction frequency of up to 6.7% was obtained on Gresshoff and Doy medium in the presence of 2 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.5 mg/l benzyladenine (BA). Formation and further proliferation of embryogenic tissue were achieved upon transfer of explants to a medium with reduced levels of growth regulators. Somatic embryos are being cultured for further development. .


HortScience ◽  
1998 ◽  
Vol 33 (3) ◽  
pp. 460e-460 ◽  
Author(s):  
Marisa F. de Oliveira ◽  
Gerson R. de L. Fortes ◽  
João B. da Silva

The aim of this work was to evaluate the organogenesis of Marubakaido apple rootstock under different aluminium concentratons. The explants were calli derived from apple internodes treated with either 2,4-dichlorophenoxyacetic acid or pichloram at 0.5 and 1.0 μM and under five different aluminium concentrations (0, 5, 10, 15, 20 mg/L). These calli were then treated with aluminium at 0, 5, 10, 15, and 20 mg/L. It was observed shoot regeneration only for those calli previously treated with pichloram. There were no significant difference among the aluminium concentrations.


HortScience ◽  
1998 ◽  
Vol 33 (3) ◽  
pp. 483a-483
Author(s):  
Roy N. Keys ◽  
Dennis T. Ray ◽  
David A. Dierig

Guayule (Parthenium argentatum Gray, Asteraceae) is a latex-producing perennial desert shrub that is potentially of economic importance as an industrial crop for the desert Southwest. It is known to possess complex reproductive modes. Diploids are predominantly sexual and self-incompatible, while polyploids show a range of apomictic potential and self-compatibility. This paper describes the development of a relatively rapid and simple technique for characterizing reproductive modes of breeding lines of P. argentatum. Initial field experiments were based on an auxin test used successfully to characterize reproductive mode in the Poaceae. The application of 2,4-dichlorophenoxyacetic acid inhibited embryo formation in P. argentatum, but this was not the case with other auxins tested. Results of field experiments were ambiguous because: 1) the floral structure of P. argentatum is such that auxins might not have penetrated to the ovules, and 2) there was potential self-fertilization by pollen released within isolation bags. Therefore, in vitro culture of flower heads was tested because it provided much better control of environmental conditions, growth regulator application, and pollen release. Auxin alone, or in combination with gibberellic acid or kinetin, inhibited parthenogenesis in vitro. Embryo production did not vary using two substantially different nutrient media. In vitro flower head culture using a (Nitsch and Nitsch) liquid nutrient medium without growth regulators, enabled characterization of the reproductive mode of seven breeding lines, ranging from predominantly sexual to predominantly apomictic. The results of this technique were substantiated using RAPD analyzes of progeny arrays from controlled crosses.


1984 ◽  
Vol 62 (7) ◽  
pp. 1393-1397 ◽  
Author(s):  
M. D. Zhou ◽  
T. T. Lee

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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