Manifestation of embryogenic potential in culture of zygotic embryos of Quercus robur L.

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-1 glutamine, proline and casein hydrolysate and supplemented with 2,4-D (1,0-2,0 mg•l-1) and BAP (0,5-1,0 mg•l-1). 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-1) or on medium with ABA and GA3 (each 0,2 mg•l-1) 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-1 BAP when cultures were mantained at 2oC for 4 weeks. Only 8 somatic embryos developed into plantlets. Their transplantation to in vivo conditions was unsuccessful.

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Somatic Embryogenesis using Immature Zygotic Embryos from Developing Fruits of Papaya (Carica papaya)

HortScience ◽

10.21273/hortsci.30.4.783e ◽

1995 ◽

Vol 30 (4) ◽

pp. 783E-783

Author(s):

S.K. Dhir ◽

U.L. Yadava

Keyword(s):

Somatic Embryogenesis ◽

Somatic Embryos ◽

Carica Papaya ◽

Induction Medium ◽

Zygotic Embryos ◽

Synthetic Seed ◽

Factors Affecting ◽

Secondary Embryos ◽

Potential Use

An efficient protocol has been developed for the in vitro multiplication of papaya (Carica papaya L.) through somatic embryogenesis utilizing immature zgotic embryos. Somatic embryos were initiated on MS basel media supplemented with 5 mg·liter–1 2,4-D, 400 mg·liter–1 glutamine, and 6% sucrose. After culturing for 2 months, 65% of the explants became highly embryogenic. Each explant produced 50 to 80 embryos in 4 months on culture induction medium. Frequency of embryogenesis was increased (75 to 150 somatic embryos on 80% explants) upon supplementing medium with 4% maltose as a carbon source and 100 mg·liter–1 L-asparagine. The embryogenic callus appeared yellow and embryos at different stages of development were well-organized. On regular subculturing, these cultures continued to produce secondary embryos. Following their transfer to the hormone-free medium supplemented with 4% maltose, these embryos germinated. The somatic embryogenesis system is rapid, repetitive, and highly proliferative. Thus, this system may have a potential use in the development of synthetic seed and transgenic papaya plants. Details of important factors affecting somatic embryogenesis will be discussed.

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

Plants ◽

10.3390/plants9010003 ◽

2019 ◽

Vol 9 (1) ◽

pp. 3 ◽

Cited By ~ 2

Author(s):

Xiuxia Ren ◽

Ya Liu ◽

Byoung Ryong Jeong

Keyword(s):

Somatic Embryogenesis ◽

Somatic Embryos ◽

Zygotic Embryo ◽

Zygotic Embryos ◽

Ms Medium ◽

Hypocotyl Explants ◽

Cotyledon Explants ◽

Embryo Induction ◽

Naphthylacetic Acid ◽

Better Than

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.

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COMPARISON OF ZYGOTIC AND SOMATIC EMBRYOGENES1S IN CERCIS CANADENSIS

HortScience ◽

10.21273/hortsci.27.11.1167d ◽

1992 ◽

Vol 27 (11) ◽

pp. 1167d-1167 ◽

Cited By ~ 1

Author(s):

L.G. Buckley ◽

E.T. Graham ◽

R.N. Trigiano

Keyword(s):

Somatic Embryogenesis ◽

Somatic Embryos ◽

Zygotic Embryo ◽

Ammonium Ion ◽

Zygotic Embryos ◽

University Of Tennessee ◽

Cercis Canadensis ◽

Developmental Sequences ◽

The University ◽

Shoot Meristems

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.

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Plant Regeneration in Sorbus. pohuashanenesis Hedl by Somatic Embryogenesis

Advanced Materials Research ◽

10.4028/www.scientific.net/amr.183-185.1462 ◽

2011 ◽

Vol 183-185 ◽

pp. 1462-1466

Author(s):

Ling Yang ◽

Yu Hua Li ◽

Hai Long Shen

Keyword(s):

Somatic Embryogenesis ◽

Plant Regeneration ◽

Somatic Embryos ◽

Casein Hydrolysate ◽

Zygotic Embryos ◽

Ms Medium ◽

Immature Zygotic Embryos

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.

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GENE EXPRESSION IN PECAN SOMATIC EMBRYOS

HortScience ◽

10.21273/hortsci.26.6.726a ◽

1991 ◽

Vol 26 (6) ◽

pp. 726A-726 ◽

Cited By ~ 2

Author(s):

Amnon Levi ◽

Hazel Y. Wetzstein ◽

Glen A. Galau

Keyword(s):

Gene Expression ◽

Somatic Embryos ◽

Zygotic Embryo ◽

Germination Rate ◽

Expression Patterns ◽

Gene Expression Patterns ◽

Zygotic Embryos ◽

Embryo Germination ◽

Coordinate Gene Expression ◽

Somatic Embryogenic

Repetitive somatic embryogenic lines of pecan (Carya illinoensis) were obtained and subcultured on basal WPM, following a one week induction of zygotic embryo tissue on modified WPM with 6 mg/L NAA. Gene expression of somatic embryos has been studied and compared with that occurring in zygotic embryos. Somatic embryos simultaneously expressed mRNA classes that are specific to each of the zygotic embryo cotyledon (Cot), maturation (Mat), and post abscission stages (Late embryogenesis, Lea). Somatic embryos exhibiting such multiple, nonregulated gene expression patterns have a low germination rate. Treatments found to enhance embryo germination (cold and desiccation) may be effective in part, by modifying gene expression patterns. Some of the Cot and Mat mRNA classes decreased following such treatments, while Lea mRNAs were not effected. Cold and desiccation treatments appear to coordinate gene expression in pecan somatic embryos, which might be associated with embryo germination.

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Histology of Somatic Embryogenesis in Regal Geranium

Journal of the American Society for Horticultural Science ◽

10.21273/jashs.119.3.648 ◽

1994 ◽

Vol 119 (3) ◽

pp. 648-651 ◽

Cited By ~ 5

Author(s):

D.P.M. Wilson ◽

J.A. Sullivan ◽

A.A. Marsolais ◽

M.J. Tsujita

Keyword(s):

Time Series ◽

Somatic Embryogenesis ◽

Strong Evidence ◽

Somatic Embryos ◽

Zygotic Embryo ◽

Zygotic Embryogenesis ◽

Parenchyma Cells

The origin and development of somatic embryos from petiole sections of Regal geranium (Pelargonium ×domesticum Bailey `Madame Layal') were studied using time-series sections at days 0, 4, 8, 14, and 24. Somatic embryos originated as early as day 4 of culture. The proembryo stage resembled that of a zygotic embryo and the somatic embryos developed through the globular, heart-torpedo, and cotyledonous stages characteristic of in vivo zygotic embryogenesis. A suspensor-like structure was observed with some somatic embryos but this was not consistent. Strong evidence is presented to suggest that somatic embryos arose from single subepidermal parenchyma cells.

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Effect of 2, 4-D and Picloram on Somatic Embryogenesis in Carica papaya var. P-7-9

Plant Tissue Culture and Biotechnology ◽

10.3329/ptcb.v29i1.41976 ◽

2019 ◽

Vol 29 (1) ◽

pp. 25-32

Author(s):

Kulbhushan Chaudhary ◽

Jai Prakash

Keyword(s):

Somatic Embryogenesis ◽

High Frequency ◽

Somatic Embryos ◽

Carica Papaya ◽

Zygotic Embryos ◽

Shoot Induction ◽

Free Medium ◽

Embryo Germination ◽

Shoot Bud ◽

Half Strength

Immature zygotic embryos of Carica papaya L. var. P-7-9 were inoculated on half-strength MS with different concentrations of 2,4-D and picloram alone and their combinations. The highest induction (80%) of somatic embryogenesis was MS supplemented with 2,4-D (7.0 mg/l) followed by 2,4-D (4.0 mg/l) + picloram (1.0 mg/l) but no embryogenesis was observed in auxin-free medium. The addition of osmoticum such as PEG and ABA significantly increased the maturation of somatic embryos. BAP, TDZ and NAA were used for shoot induction. Combination of 2.0 mg/l BAP and 0.5 mg/l NAA was found significantly better for shoot bud proliferation and 70% multiplication. The percentage of germinating embryos was enhanced significantly when the MS fortified with GA3 (0.5 mg/l). However, the high frequency of embryo germination and plantlets formation were obtained with 2,4-D (4.0 mg/l) + picloram (1.0 mg/l). Plant Tissue Cult. & Biotech. 29(1): 25-32, 2019 (June)

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Induction of somatic embryogenesis in Pinus heldreichii culture

Archives of Biological Sciences ◽

10.2298/abs0703199s ◽

2007 ◽

Vol 59 (3) ◽

pp. 199-202 ◽

Cited By ~ 3

Author(s):

Dragana Stojicic ◽

Branka Uzelac ◽

Dusica Janosevic ◽

Ljubinka Culafic ◽

Snezana Budimir

Keyword(s):

Somatic Embryogenesis ◽

Somatic Embryos ◽

Zygotic Embryo ◽

Embryogenic Tissue ◽

Zygotic Embryos ◽

Dichlorophenoxyacetic Acid ◽

Immature Zygotic Embryos ◽

Induction Frequency ◽

2,4 Dichlorophenoxyacetic Acid ◽

Further Development

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. .

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Somatic embryogenesis and plantlet regeneration from embryos isolated from stored seeds of Picea glauca

Canadian Journal of Botany ◽

10.1139/b90-033 ◽

1990 ◽

Vol 68 (2) ◽

pp. 236-242 ◽

Cited By ~ 64

Author(s):

F. M. Tremblay

Keyword(s):

Somatic Embryogenesis ◽

Embryogenic Callus ◽

Picea Glauca ◽

Somatic Embryos ◽

Germination Rate ◽

Casein Hydrolysate ◽

White Spruce ◽

Zygotic Embryos ◽

Dichlorophenoxyacetic Acid ◽

2,4 Dichlorophenoxyacetic Acid

White spruce (Picea glauca) embryogenic callus was obtained using 3- to 11-year-old seeds as a source of zygotic embryos. They were cultured on half-strength Litvay's medium supplemented with 10 μM 2,4-dichlorophenoxyacetic acid, 5 μM benzylaminopurine, 1 g/L casein hydrolysate, 500 mg/L glutamine, and 1% sucrose. The frequency of induction of embryogenic callus was significantly improved by incubation at 25 °C and by a 4-h imbibition of the seeds. The yield of embryogenic callus was significantly affected by the geographic provenance of the seeds and by their number of years in storage. A significant correlation was also found between the yield of embryogénie callus and the percentage of germination of the seedlot used. Even after 11 years of storage, 40% of the zygotic embryos could produce an embryogenic callus when dissected from seeds with a high germination rate. Somatic embryos were matured after transfer onto an embryo development medium composed of the same medium but including 6% sucrose, 1 μM 2,4-dichlorophenoxyacetic acid, and 5 μM kinetin. The somatic embryos developed further under in vitro conditions and were then transplanted into soil. The somatic embryoderived plantlets established in the greenhouse were similar to control plantlets obtained from germinated seeds. Mature embryos from stored seeds were shown to constitute a valuable source for white spruce somatic embryogenesis.

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Induction of Somatic Embryogenesis in Vulnerable Medicinal Plant Hedychium spicatum Buch-Ham ex Smith

Plant Tissue Culture and Biotechnology ◽

10.3329/ptcb.v23i2.17506 ◽

2014 ◽

Vol 23 (2) ◽

pp. 147-155 ◽

Cited By ~ 1

Author(s):

Dinesh Giri ◽

Sushma Tamta

Keyword(s):

Somatic Embryogenesis ◽

Medicinal Plant ◽

Somatic Embryos ◽

Synthetic Seeds ◽

Ex Situ ◽

Secondary Embryogenesis ◽

Zygotic Embryos ◽

High Concentration ◽

Cotyledon Explants ◽

Short Period

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)

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