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

1990 ◽  
Vol 68 (2) ◽  
pp. 236-242 ◽  
Author(s):  
F. M. Tremblay

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.

1987 ◽  
Vol 65 (4) ◽  
pp. 656-659 ◽  
Author(s):  
Inger Hakman ◽  
Larry C. Fowke

Embryogenic callus was initiated from immature embryos of Picea glauca (white spruce) and Picea mariana (black spruce) cultured on defined media supplemented with 2,4-dichlorophenoxyacetic acid (1 × 10−5 M), N6-benzyladenine (5 × 10−6 M), and 1% sucrose. Seeds from cones stored at 4 °C for up to 3 months yielded embryogenic callus. Much higher frequencies of embryogenic callus were obtained from white spruce than from black spruce. Embryogenic callus contained loosely organized cells and somatic embryos of various sizes. The embryos consisted of a cluster of tiny dividing cells (embryonic region) with attached large vacuolated cells (suspensor region). Upon subculture of embrogenic callus to media either lacking growth regulators or with reduced concentrations (5 × 10−7 M, 2,4-dichlorophenoxyacetic acid and 5 × 10−6 M N6-benzyladenine) somatic embryos could be stimulated to develop into plantlets.


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.


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.


2001 ◽  
Vol 44 (3) ◽  
pp. 247-255 ◽  
Author(s):  
Juliana A. Fernando ◽  
Murilo Melo ◽  
Marli K. M. Soares ◽  
Beatriz Appezzato-da-Glória

Mature zygotic embryos of Carica papaya L. ‘Sunrise Solo’ were used as explants for embryogenesis induction. The explants were inoculated on Murashige and Skoog culture medium supplemented with 2 mg.L-1 2,4-dichlorophenoxyacetic acid and incubated in darkness at 25+2°C. Histological analysis of callogenesis and somatic embryogenesis indicated occurrence of direct and indirect somatic embryogenesis development. Direct somatic embryo formation was observed from hypocotyledonary epidermic cells only from explant 18 days after inoculation. Somatic embryos formed indirectly were originated from embryogenic superficial cells of pre-embryonic complexes located on peripherical and on internal cell layers of callus 49 days after inoculation. Diverse morphological differences including disformed embryos were observed among the somatic embryos.


Biologia ◽  
2010 ◽  
Vol 65 (5) ◽  
Author(s):  
Shaoyu Chen ◽  
Shanna Chen ◽  
Fang Chen ◽  
Tao Wu ◽  
Yinbin Wang ◽  
...  

AbstractSomatic embryogenesis (SE) was successfully induced from mature zygotic embryos of seven families of Picea likiangensis (Franch.) Pritz after 20 weeks culture on initiation medium. Three basal media (one-half strength LM medium, one-half strength LP medium and improved LP medium) with different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-benzyladenine (6-BA) were tested but only one-half strength LM medium supplemented with 2,4-D and 6-BA was successful for the embryogenic cultures (EC) initiation. The initiation frequencies of EC varied greatly from different families when culturing on the same initiation medium. The highest frequency (41.3%) was induced from one of the families on one-half strength LM medium supplemented with 3 mg L−1 2,4-D and 1.5 mg L−1 6-BA and 16.83% on average for seven families. EC were subcultured and proliferated on the same medium as the initiation one every 10 days. 3 lines of EC induced from the same family were applied in maturation experiment. Cotyledonary somatic embryos were observed after EC were transferred to maturation media of one-half strength LM medium containing 20-80 mg L−1 abscisic acid and 7.5% polyethylene glycol (PEG-4000). However, one-half strength LM medium supplemented with 40 mg L−1 or 60 mg L−1 ABA and 7.5% PEG gave the best maturation and the 3 lines showed different ability in maturation. Over 80% cotyledonary somatic embryos germinated normally on DCR medium containing 0.2% activated carbon. The success on SE induction of the species has provided an effective clonal propagation method for this important tree’s genetic improvement.


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.


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. .


2017 ◽  
Vol 17 (1) ◽  
pp. 9
Author(s):  
Yosi Zendra Joni ◽  
Riry Prihatini ◽  
Darda Efendi ◽  
Ika Roostika

<p>Somatic embryogenesis is a technique for regenerating embryos derived from somatic cells of various plant species. This technique along with the utilization of plant growth regulator (PGR) might benefit for mass propagation and improvement of plant species through biotechnological tools. The study aimed to determine the effect of different plant growth regu-lators, namely 6-benzyladenine (BA) and thidiazuron (TDZ) on the embryogenic callus induction as well as casein hydrolysate and malt extract on the somatic embryo development of mangosteen. The explants used were in vitro young stems of mangosteen clone Leuwiliang. This study consisted of two experiments, namely induction of embryogenic callus and formation of somatic embryo. The first experiment was arranged as factorial in a completely randomized design with BA (0 and 0.7 mg l-1) as the first factor and TDZ (0, 0.1, 0.5 and 1.0 mg l-1) as the second factor. The second experiment consisted of four treatments, i.e. casein hydrolysate and malt extract at the rate of 500 and 1,000 mg l-1. The results showed that the best medium for embryogenic callus induction was MS supplemented with 0.1 mg l-1 TDZ, which resulted semifriable calli. Casein hydrolysate and malt extract could not induce the formation of somatic embryos. After two times subcultures on the same MS medium supplemented with 0.5 mg l-1 TDZ and 0.7 mg l-1 BA, a total of 33.8 somatic embryos per explant was induced. The successful somatic embryogenesis would support mangosteen breeding and in vitro mass propagation program.</p>


Plants ◽  
2022 ◽  
Vol 11 (2) ◽  
pp. 168
Author(s):  
Muhammad Ajmal Bashir ◽  
Cristian Silvestri ◽  
Amelia Salimonti ◽  
Eddo Rugini ◽  
Valerio Cristofori ◽  
...  

An efficient in vitro morphogenesis, specifically through somatic embryogenesis, is considered to be a crucial step for the application of modern biotechnological tools for genetic improvement in olive (Olea europaea L.). The effects of different ethylene inhibitors, i.e., cobalt chloride (CoCl2), salicylic acid (SA), and silver nitrate (AgNO3), were reported in the cyclic somatic embryogenesis of olive. Embryogenic callus derived from the olive immature zygotic embryos of the cultivar Leccino, was transferred to the expression ECO medium, supplemented with the ethylene inhibitors at 20 and 40 µM concentrations. Among these, the maximum number of somatic embryos (18.6) was obtained in media containing silver nitrate (40 µM), followed by cobalt chloride (12.2 somatic embryos @ 40 µM) and salicylic acid (40 µM), which produced 8.5 somatic embryos. These compounds interfered on callus traits: white friable embryogenic calli were formed in a medium supplemented with 40 µM cobalt chloride and salicylic acid; in addition, a yellow-compact embryogenic callus appeared at 20 µM of all the tested ethylene inhibitors. The resulting stimulatory action of silver nitrate among all the tested ethylene inhibitors on somatic embryogenesis, clearly demonstrates that our approach can efficiently contribute to the improvement of the current SE protocols for olive.


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.


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