scholarly journals Enhanced Somatic Embryo Induction of a Tree Peony, Paeonia ostii ‘Fengdan’, by a Combination of 6-benzylaminopurine (BA) and 1-naphthylacetic Acid (NAA)

Plants ◽  
2019 ◽  
Vol 9 (1) ◽  
pp. 3 ◽  
Author(s):  
Xiuxia Ren ◽  
Ya Liu ◽  
Byoung Ryong Jeong

Somatic embryogenesis is a preferred method for vegetative propagation due to its high propagation efficiency. In this study, zygotic embryos, cotyledons, and hypocotyls of Paeonia ostii ‘Fengdan’ were used as the explant to induce somatic embryogenesis. The results showed that a combination of 0.5 mg·L−1 thidiazuron (TDZ) and 0.5 mg·L−1 2,4-dichlorophenoxyacetic acid (2,4-D) was effective in inducing somatic embryos from the zygotic embryo and cotyledon explants. Hypocotyls only formed somatic embryos on Murashige and Skoog (MS) medium supplemented with both 0.5 mg·L−1 TDZ and 0.5 mg·L−1 1-naphthylacetic acid (NAA). Moreover, the compact callus was effectively produced from zygotic embryo, cotyledon, and hypocotyl explants in medium supplemented with a combination of 3.0 mg·L−1 6-benzylaminopurine (BA) and 1.0 mg·L−1 NAA, and then converted into somatic embryos in the same medium, and the ratio of the explants with embryo induction and number of embryos induced per explant were much higher than those induced by 0.5 mg·L−1 TDZ and either 0.5 mg·L−1 2,4-D or 0.5 mg·L−1 NAA. The MS medium was better than the woody plant medium (WPM) for inducing somatic embryos from zygotic embryo and hypocotyl explants, whereas the WPM was better than the MS medium for somatic embryogenesis induction from cotyledon explants. All of the somatic embryos developed well into mature embryos on their respective media supplemented with both 3.0 mg·L−1 BA and 1.0 mg·L−1 NAA. Overall, the protocols for indirect somatic embryogenesis from zygotic embryo, cotyledon, and hypocotyl of P. ostii ‘Fengdan’ were successfully established, which can greatly facilitate their propagation and breeding processes.

HortScience ◽  
2002 ◽  
Vol 37 (7) ◽  
pp. 1122-1123 ◽  
Author(s):  
Mohammed A.M. Aly ◽  
Bala Rathinasabapathi ◽  
Sheevani Bhalsod

Many members of the Plumbaginaceae are important flower crops wherein propagation is hindered by poor seed germination. Micropropagation via organogenesis is commercially practiced for certain Limonium species. However, somatic embryogenesis was not reported for members of the Plumbaginaceae until recently for L. bellidifolium Durmort. The induction of somatic embryogenesis from cotyledon explants in a modified Murashige and Skoog (MS) medium was examined in four other members of this family, Limonium aureum O. Kuntze, L. latifolium O. Kuntze, L. sinuatum Mill., and Armeria maritima Willd. Induction of embryogenic callus was achieved in all the species examined on MS medium supplemented with 4.5 μm 2,4-D and 88 or 118 mm sucrose. Species of the genus Limonium responded better than A. maritima Willd. in somatic embryo induction and maturation. Somatic embryos of L. aureum O. Kuntze matured readily on MS medium supplemented with 0.93 μm kinetin and 88 mm mannitol. Chemical name used: 2,4-Dichlorophenoxy acetic acid (2,4-D).


2014 ◽  
Vol 23 (2) ◽  
pp. 147-155 ◽  
Author(s):  
Dinesh Giri ◽  
Sushma Tamta

This protocol has been developed for somatic embryogenesis in Hedychium spicatum. Simultaneously, a method has also been developed for the production of synthetic seeds by using somatic embryos. Direct somatic embryos were developed on cotyledon explants of zygotic embryos on MS supplemented with high concentration of NAA (20.0 µM). Induction of secondary embryogenesis was best in 2,4-D supplemented medium fortified with activated charcoal. Germination of somatic embryos was enhanced by using GA3. Besides this, round and semi-hard beads of somatic embryos (synthetic seeds) could be produced by using 2% Na-alginate and 100 mM calcium chloride and more than 30% germination of synthetic seeds was achieved in MS. Well acclimated plants produced via somatic embryogenesis and/or synthetic seeds were transferred to field where more than 60% survived. This simple study enabled us to obtain a number of plantlets throughout the year each cycle requiring a short period of time. Besides propagation, this study provided an ex situ method for conservation of this vulnerable Himalayan species.D. O. I.http://dx.doi.org/10.3329/ptcb.v23i2.17506Plant Tissue Cult. & Biotech. 23(2): 147-155, 2013  (December)


Development ◽  
1997 ◽  
Vol 124 (10) ◽  
pp. 2049-2062 ◽  
Author(s):  
E.D. Schmidt ◽  
F. Guzzo ◽  
M.A. Toonen ◽  
S.C. de Vries

The first somatic single cells of carrot hypocotyl explants having the competence to form embryos in the presence of 2,4-dichlorophenoxyacetic acid (2,4-D) were identified using semi-automatic cell tracking. These competent cells are present as a small subpopulation of enlarged and vacuolated cells derived from cytoplasm-rich and rapidly proliferating non-embryogenic cells that originate from the provascular elements of the hypocotyl. A search for marker genes to monitor the transition of somatic into competent and embryogenic cells in established suspension cell cultures resulted in the identification of a gene transiently expressed in a small subpopulation of the same enlarged single cells that are formed during the initiation of the embryogenic cultures from hypocotyl explants. The predicted amino acid sequence and in vitro kinase assays show that this gene encodes a leucine-rich repeat containing receptor-like kinase protein, designated Somatic Embryogenesis Receptor-like Kinase (SERK). Somatic embryos formed from cells expressing a SERK promoter-luciferase reporter gene. During somatic embryogenesis, SERK expression ceased after the globular stage. In plants, SERK mRNA could only be detected transiently in the zygotic embryo up to the early globular stage but not in unpollinated flowers nor in any other plant tissue. These results suggest that somatic cells competent to form embryos and early globular somatic embryos share a highly specific signal transduction chain with the zygotic embryo from shortly after fertilization to the early globular embryo.


2014 ◽  
Vol 65 (1-2) ◽  
pp. 37-41 ◽  
Author(s):  
Maria G. Ostrolucká ◽  
Diana Krajmerová

For the initiation of somatic embryogenesis early cotyledonary stage of zygotic embryo explants (from 15th July until late August) was suitable. The highest frequency of differentiation of somatic embryos was obtained on cotyledons of zygotic embryos cultured on basal modified medium MS (with 1/2 concentration macronutrients) or WPM medium containing 500 mg•l<sup>-1</sup> glutamine, proline and casein hydrolysate and supplemented with 2,4-D (1,0-2,0 mg•l<sup>-1</sup>) and BAP (0,5-1,0 mg•l<sup>-1</sup>). The development of somatic embryos was direct and indirect and the process was continuous over a long period. Primary somatic embryos were able to produce secondary embryos. Repetitive somatic embryogenesis led to the proliferation of a large number of new somatic embryos on their cotyledons, hypocotyl or radicula. The process of embryo differentation is asynchronous - various stages of somatic embryos could be observed in embryogenic culture. A somatic embryo conversion was rare on tested media. Embryo germination occured on medium containing BAP (0,1 mg•l<sup>-1</sup>) or on medium with ABA and GA<sub>3</sub> (each 0,2 mg•l<sup>-1</sup>) after a previous culture on WPM medium without plant growth regulators supplemented with sorbitol (6%). The embryo germination occurred also on WPM medium with 0.2 mg•l<sup>-1</sup> BAP when cultures were mantained at 2<sup>o</sup>C for 4 weeks. Only 8 somatic embryos developed into plantlets. Their transplantation to <em>in vivo</em> conditions was unsuccessful.


HortScience ◽  
1992 ◽  
Vol 27 (11) ◽  
pp. 1167d-1167 ◽  
Author(s):  
L.G. Buckley ◽  
E.T. Graham ◽  
R.N. Trigiano

Zygotic and somatic embryos are purported to follow similar developmental sequences, but few investigations have thoroughly compared the two processes. Developing pods of Cercis canadensis L. (redbud) were collected from trees on the Knoxville campus of the University of Tennessee once or twice per week from 28 March to 8 August 1991. At least 10 ovules/sample date were fixed in FAA to evaluate zygotic embryo ontogeny. A minimum of 40 ovules/sample date were aseptically excised and placed on SH medium supplemented with 9.0 μM 2,4-D and 5 mM ammonium ion to initate somatic embryogenesis. Zygotic and somatic embryos were prepared for histological examination using standard paraffin techniques. Somatic embryos developed primarily from cotyledons and epicotyls of zygotic embryos mat were cultured between 6 June and 19 July. Somatic and zygotic embryos were subtended by multiseriate suspensors and progressed through recognizable globular, cordate and cotyledonary stages of development. Cotyledon morphology was similar for both embryo types. However, many somatic embryos failed to differentiate dome-shaped shoot meristems exhibited by their zygotic counterparts.


2011 ◽  
Vol 14 (3) ◽  
pp. 23-29
Author(s):  
Phuc Thanh Vo ◽  
Tien Thi Thuy Le

Callus was initiated from hypocotyls and cotyledons explants of chilli Capsicum sp. in vitro on MS medium with 0,5 mg/l kinetin and 2,4-D /NAA (1,0; 1,5; 2,0; 2,5 and 3,0 mg/l). Callus from cotyledon explants was induced in the dark better than in the light, whereas callus from hypocotyl explants was initiated in the light better than in the dark. Callus was more friable and grew faster on medium with 2,4-D and kinetin. MS medium with 3,0 mg/l 2,4-D and 0,5 mg/l kinetin was optimal for the growth of callus from cotyledon explants. Besides, callus from hypocotyl explants grew best on MS medium with 1,5 mg/l 2,4-D and 0,5 mg/l kinetin. Capsaicinoid from callus which was determined by thin layer chromatography was recognized in all treatment experiments.


2011 ◽  
Vol 183-185 ◽  
pp. 1462-1466
Author(s):  
Ling Yang ◽  
Yu Hua Li ◽  
Hai Long Shen

Somatic embryogenesis was obtained by using immature zygotic embryos of S. pohuashanesis as explants and emblings were obtained. For induction of somatic embryos, immature zygotic embryos which 30 days old after pollination were cultured on solid MS medium with 1.0 mg•L-1 NAA, 0.1 mg•L-1 6-BA, 500 mg•L-1casein hydrolysate (CH) and 40 g•L-1 sucrose . Inducted somatic embryos were cultured in solid MS medium containing 500 mg•L-1CH and 40 g•L-1 sucrose. After 30 days of culture, many normal cotyledonary embryos were produced. Plantlets were regenerated when somatic embryos were transferred to MS medium with 30 g•L-1 sucrose. The somatic embryos germinated at a germination frequency of approximately 80%, but rate of the plantlets that successfully acclimated and continued growing was 40% in the greenhouse.


2011 ◽  
Vol 2011 ◽  
pp. 1-8 ◽  
Author(s):  
Premananda Das

Somatic embryogenesis was achieved in four leguminous tree species, that is, Acacia catechu, Acacia arabica, Hardwickia binata, and Dalbergia sissoo using immature zygotic embryos as explants on Murashige and Skoog (MS) medium supplemented with 0.25–1.0 mg/l Kn (kinetin) and 2.0–3.0 mg/l 2,4-D (2,4-dichlorophenoxyacetic acid) or NAA (1-napthaleneacetic acid) and 3% sucrose. MS medium containing 2.0 mg/l 2,4-D and 1.0–1.5 mg/l Kn was noted to be most effective in inducing friable embryogenic callus (FEC); the number of somatic embryos per culture varied in MS medium supplemented with 1.0–2.0 mg/l 2,4-D or NAA and 0.25–1.5 mg/l kinetin. The maximum number of somatic embryos was obtained in MS medium containing 1.5–2.0 mg/l 2,4-D or NAA and 1.0–1.5 mg/l kinetin; proliferation of embryogenic calli was enhanced in cultures having 1.0–2.0 mg/l 2,4-D, 1.0–1.5 mg/l kinetin, and 400–600 mg/l L-Proline. The somatic embryos in various shapes and sizes after the first subculture on MS medium supplemented with 0.1 mg/l IAA and 0.25 mg/l BA; developed shoots and rooted in strength MS medium supplemented with 0.1 mg/l IBA or IAA. The somatic embryo-derived plantlets were transferred to the field after being hardened in the climate-controlled hardening chamber.


2002 ◽  
Vol 24 (3) ◽  
pp. 601-603 ◽  
Author(s):  
Ana da Silva Ledo ◽  
Osmar Alves Lameira ◽  
Abdellatif Kemaleddine Benbadis ◽  
Ilmarina Campos de Menezes ◽  
Maria do Socorro Padilha de Oliveira ◽  
...  

The aim of this work was to study the morphogenetic responses of zygotic embryos of açai palm (Euterpe oleracea Mart.) submitted to several conditions of in vitro culture. Several research experiments were conducted, in laboratory, using vegetable material collected from açai palm plants at Embrapa Amazon Oriental, Belém-PA, Brazil. It was possible to verify the expression of a direct, repetitive and no-synchronized model of somatic embryogenesis in mature zygotic embryos cultivated in primary MS medium supplemented with 2,4-D (339.36 muM) and transferred to a secondary MS medium in the presence of NAA (0.537 muM) and 2iP (12.30 muM). The conversion of somatic embryos into seedlings was reached after 210 days with the transfer of the cultures to a third medium with sucrose and mineral salts concentrations reduced to a half, without growth regulators.


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).


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