scholarly journals Somatic embryogenesis of selected spruce species (Picea abies, P. omorika, P. pungens 'Glauca' and P. breweriana)

2011 ◽  
Vol 77 (3) ◽  
pp. 189-199 ◽  
Author(s):  
Teresa Hazubska-Przybył ◽  
Krystyna Bojarczyk
Keyword(s):  
Somatic Embryogenesis ◽  
Picea Abies ◽  
Sucrose Concentration ◽  
Medium Composition ◽  
Embryogenic Tissue ◽  
Zygotic Embryos ◽  
Embryogenic Cell ◽  
Nursery Production ◽  
Embryogenic Cell Lines

Somatic embryogenesis was studied in four spruce species (<em>Picea abies</em>, <em>P. omorika</em>, <em>P. pungens</em> 'Glauca' and <em>P. brewenana</em>) to determine if this method can be used for in vitro propagation of coniferous trees. The highest frequency of initiation of embryogenic tissue was obtained when mature zygotic embryos were used as explants. It ranged then from 10.8% (<em>P. brewenana</em>) to 23.75% (<em>P. omorika</em> and <em>P. pungens</em> 'Glauca'). The frequency of embryogenic tissue initiation was strongly affected by medium composition, i.e. addition of appropriate auxins (2,4-D, NAA, Picloram) and sucrose concentration (10-20 g<sup>-1</sup>"1). A lower frequency was obtained in <em>Picea omorika</em> (10%) when megagametophytes (endosperms with immature zygotic embryos) were used as explants. No emryogenic tissue was produced from hypocotyls, cotyledons and needles. A satisfactory frequency was achieved with the use of somatic embryos of <em>Picea abies</em> (30%). The proliferation of embryogenic cell lines of spruces was affected by medium type. The experiments resulted in production of somatic plantlets of <em>P. abies</em> and <em>P. omorika</em>. This enables the application of this method of spruce micropropagation for genetic and breeding research or for nursery production.

scholarly journals Pinus canariensis plant regeneration through somatic embryogenesis

2020 ◽  
Vol 29 (1) ◽  
pp. eSC05
Author(s):  
Ander Castander-Olarrieta ◽  
Paloma Moncaleán ◽  
Itziar A. Montalbán
Keyword(s):  
Somatic Embryogenesis ◽  
Cell Lines ◽  
Somatic Embryos ◽  
Embryogenic Tissue ◽  
Established Cell Line ◽  
Zygotic Embryos ◽  
Pinus Canariensis ◽  
Embryogenic Cell ◽  
Embryogenic Cell Lines ◽  
Established Cell

Aim of the study: To develop an efficient method to regenerate plants through somatic embryogenesis of an ecologically relevant tree species such as Pinus canariensis.Area of study: The study was conducted in the research laboratories of Neiker-Tecnalia (Arkaute, Spain).Material and methods: Green cones of Pinus canariensis from two collection dates were processed and the resulting immature zygotic embryos were cultured on three basal media. The initiated embryogenic tissues were proliferated testing two subculture frequencies, and the obtained embryogenic cell lines were subjected to maturation. Germination of the produced somatic embryos was conducted and acclimatization was carried out in a greenhouse under controlled conditions.Main results: Actively proliferating embryogenic cell lines were obtained and well-formed somatic embryos that successfully germinated were acclimatized in the greenhouse showing a proper growth.Research highlights: This is the first report on Pinus canariensis somatic embryogenesis, opening the way for a powerful biotechnological tool for both research purposes and massive vegetative propagation of this species.Keywords: acclimatization; Canary Island pine; micropropagation; embryogenic tissue; somatic embryo.Abbreviations used: embryogenic tissue (ET); established cell line (ECL);  somatic embryogenesis (SE); somatic embryos (Se’s).

scholarly journals Somatic Embryogenesis in Arabidopsis thaliana Is Facilitated by Mutations in Genes Repressing Meristematic Cell Divisions

Genetics ◽  
1998 ◽  
Vol 149 (2) ◽  
pp. 549-563
Author(s):  
Andreas P Mordhorst ◽  
Keete J Voerman ◽  
Marijke V Hartog ◽  
Ellen A Meijer ◽  
Jacques van Went ◽  
...  
Keyword(s):  
Somatic Embryogenesis ◽  
Liquid Medium ◽  
Cell Cultures ◽  
Egg Cell ◽  
Zygotic Embryos ◽  
Additive Effects ◽  
Additional Advantage ◽  
Embryogenic Cell ◽  
Immature Zygotic Embryos

Abstract Embryogenesis in plants can commence from cells other than the fertilized egg cell. Embryogenesis initiated from somatic cells in vitro is an attractive system for studying early embryonic stages when they are accessible to experimental manipulation. Somatic embryogenesis in Arabidopsis offers the additional advantage that many zygotic embryo mutants can be studied under in vitro conditions. Two systems are available. The first employs immature zygotic embryos as starting material, yielding continuously growing embryogenic cultures in liquid medium. This is possible in at least 11 ecotypes. A second, more efficient and reproducible system, employing the primordia timing mutant (pt allelic to hpt, cop2, and amp1), was established. A significant advantage of the pt mutant is that intact seeds, germinated in 2,4-dichlorophenoxyacetic acid (2,4-D) containing liquid medium, give rise to stable embryonic cell cultures, circumventing tedious hand dissection of immature zygotic embryos. pt zygotic embryos are first distinguishable from wild type at early heart stage by a broader embryonic shoot apical meristem (SAM). In culture, embryogenic clusters originate from the enlarged SAMs. pt somatic embryos had all characteristic embryo pattern elements seen in zygotic embryos, but with higher and more variable numbers of cells. Embryogenic cell cultures were also established from seedling, of other mutants with enlarged SAMs, such as clavata (clv). pt clv double mutants showed additive effects on SAM size and an even higher frequency of seedlings producing embryogenic cell lines. pt clv double mutant plants had very short fasciated inflorescence stems and additive effects on the number of rosette leaves. This suggests that the PT and CLV genes act in independent pathways that control SAM size. An increased population of noncommitted SAM cells may be responsible for facilitated establishment of somatic embryogenesis in Arabidopsis.

Evaluation of somatic embryogenesis for clonal propagation of western white pine

2000 ◽  
Vol 30 (12) ◽  
pp. 1867-1876 ◽  
Author(s):  
R E Percy ◽  
K Klimaszewska ◽  
D R Cyr
Keyword(s):  
Somatic Embryogenesis ◽  
Somatic Embryos ◽  
Clonal Propagation ◽  
Embryogenic Tissue ◽  
Zygotic Embryos ◽  
Pinus Monticola ◽  
White Pine ◽  
Western White Pine ◽  
Embryogenic Lines

A multiyear program was undertaken to develop a somatic embryogenesis system for clonal propagation of western white pine (Pinus monticola Dougl.). Developing seeds were used to initiate embryogenic lines from families used in blister-rust (Cronartium ribicola J.C. Fisch.) resistance breeding programs in British Columbia. The most responsive seeds contained zygotic embryos ranging in development from late cleavage polyembryony to the early dominance stage. Overall, 14 of 15 open-pollinated families produced embryogenic lines. The best results (0.8-6.7% initiation) were obtained using modified Litvay medium with 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-benzyladenine (BA) at 2.25 µM. Proliferation of embryogenic tissue was enhanced by culturing tissue as a thin layer on filter paper supports. Approximately 300 lines representing 18 open- and control-pollinated families were cryopreserved. The highest number of mature somatic embryos was obtained on maturation medium containing 120 µM abscisic acid, 180 mM sucrose, and 1.0% gellan gum. Of 61 lines tested on this medium, 77% produced mature somatic embryos. In vitro germination and early growth occurred at a high frequency (90-95%), and plants from 45 genotypes were subsequently transferred to a greenhouse.

scholarly journals Physiological and Structural Aspects of In Vitro Somatic Embryogenesis in Abies alba Mill

Forests ◽  
2020 ◽  
Vol 11 (11) ◽  
pp. 1210
Author(s):  
Terezia Salaj ◽  
Katarina Klubicová ◽  
Bart Panis ◽  
Rony Swennen ◽  
Jan Salaj
Keyword(s):  
Somatic Embryogenesis ◽  
Cell Lines ◽  
Somatic Embryos ◽  
Abies Alba ◽  
Embryogenic Tissue ◽  
Zygotic Embryos ◽  
Peg 4000 ◽  
Initiation Frequency ◽  
Embryogenic Tissues

Initiation of somatic embryogenesis from immature zygotic embryos, long-term maintenance of embryogenic tissue in vitro or by cryopreservation, as well as maturation, of somatic embryos of Abies alba Mill. are reported in this study. For the initiation of embryogenic tissues, a DCR medium containing different types of cytokinins (1 mg.L−1) were tested. During three consecutive years, 61 cell lines were initiated out of 1308 explants, with initiation frequencies ranging between 0.83 and 13.33%. The type of cytokinin had no profound effect on the initiation frequency within one given year. Microscopic observations revealed presence of bipolar somatic embryos in all initiated embryogenic tissues. Besides the typical bipolar somatic embryos, huge polyembryonal complexes, as well as “twin” embryos, were observed. Maturation of somatic embryos occurred on a DCR medium supplemented by abscisic acid (10 mg.L−1), polyethylene glycol (PEG-4000, 7.5%) and 3% maltose. The maturation capacity was cell-line dependent. All of the four tested cell lines produced cotyledonary somatic embryos, though at different quantities, of 16 to 252 per g of fresh weight. After germination, seedlings developed, but their further growth soon stopped after the formation of a resting bud. Altogether, seven cell lines were cryopreserved, using the slow-freezing technique. After rewarming, all tested cell lines showed regrowth rates between 81.8 and 100%.

scholarly journals In vitro germination and embryogenic competence acquisition of Euterpe edulis Martius immature zygotic embryos

2012 ◽  
Vol 12 (3) ◽  
pp. 171-178 ◽  
Author(s):  
Cleber Witt Saldanha ◽  
Maisa Pimentel Martins-Corder
Keyword(s):  
Somatic Embryogenesis ◽  
Calcium Chloride ◽  
Somatic Embryos ◽  
Sucrose Concentration ◽  
Zygotic Embryos ◽  
Palm Tree ◽  
In Vitro Germination ◽  
Embryogenic Competence ◽  
Immature Zygotic Embryos

This study evaluated different aspects of in vitro germination and embryogenic competence of immature zygotic embryos of E. edulis. The embryos germinated on full or half-strength MS (MS or MS/2) medium combined with sucrose (20, 30 and 40 g L-1). The effect of calcium chloride concentrations (0, 2, 4, 8, and 12 mM) on the induction of somatic embryogenesis was tested. The embryos were germinated on MS or MS/2. Germination of zygotic embryos and the number of roots per plantlet were not affected by the culture medium and sucrose concentration. Plantlet height and fresh weight were influenced by both; the difference was greatest in MS medium with 40 g L-1 sucrose. The induction of somatic embryogenesis was not influenced by doses of calcium chloride, whereas the number of somatic embryos formed was affected. The germination capacity of somatic embryos of heart-of-palm tree was not influenced by the media tested.

scholarly journals Somatic embryogenesis as an alternative for in vitro multiplication of Butia odorata from mature zygotic embryos

2020 ◽  
Vol 92 (suppl 1) ◽  
Author(s):  
SAMANTA S. DE CAMPOS ◽  
JONNY E. SCHERWINSKI-PEREIRA ◽  
REGINA B. BERND ◽  
CLAUDIMAR S. FIOR ◽  
SERGIO F. SCHWAZ
Keyword(s):  
Somatic Embryogenesis ◽  
Zygotic Embryos ◽  
In Vitro Multiplication

scholarly journals Micropropagation of Cyclopia genistoides, an Endemic South African Plant of Economic Importance

2012 ◽  
Vol 67 (1-2) ◽  
pp. 65-76 ◽  
Author(s):  
Adam Kokotkiewicz ◽  
Maria Luczkiewicz ◽  
Anna Hering ◽  
Renata Ochocka ◽  
Krzysztof Gorynski ◽  
...  
Keyword(s):  
South African ◽  
Sucrose Concentration ◽  
Shoot Multiplication ◽  
Potassium Nitrate ◽  
Economic Importance ◽  
Medium Composition ◽  
Root Induction ◽  
Multiplication Rate ◽  
Shoot Cultures

An efficient micropropagation protocol of Cyclopia genistoides (L.) Vent., an indigenous South African shrub of economic importance, was established. In vitro shoot cultures were obtained from shoot tip fragments of sterile seedlings cultured on solid Schenk and Hildebrandt (SH) medium supplemented with 9.84 μM 6-(γ,γ-dimethylallylamino)purine (2iP) and 1.0 μM thidiazuron (TDZ). Maximum shoot multiplication rate [(8.2 ± 1.3) microshoots/explant)] was observed on this medium composition. Prior to rooting, the multiplied shoots were elongated for 60 days (two 30-days passages) on SH medium with one-half sucrose concentration, supplemented with 4.92 μM indole-3-butyric acid (IBA). The rooting of explants was only possible in the case of the elongated shoots. The highest root induction rate (54.8%) was achieved on solid SH medium with one-half sucrose and one-half potassium nitrate and ammonium nitrate concentration, respectively, supplemented with 28.54 μM indole-3-acetic acid (IAA) and 260.25 μM citric acid. The plantlets were acclimatized for 30 days in the glasshouse, with the use of peat/gravel/perlite substrate (1:1:1). The highest acclimatization rate (80%) was obtained for explants rooted with the use of IAA-supplemented medium. The phytochemical profile of the regenerated plants was similar to that of the reference intact plant material. HPLC analyses showed that C. genistoides plantlets obtained by the micropropagation procedure kept the ability to produce xanthones (mangiferin and isomangiferin) and the fl avanone hesperidin, characteristic of wild-growing shrubs.

scholarly journals Blue sky’s the limit? Somatic embryogenesis as a means of propagating recalcitrant blue spruce (Picea pungens) cultivar Hoopsii

2019 ◽  
Author(s):  
Jordan Demone ◽  
Jingqin Mao ◽  
Shen Wan ◽  
Maryam Nourimand ◽  
Äsbjörn Erik Hansen ◽  
...  
Keyword(s):  
Somatic Embryogenesis ◽  
Somatic Embryos ◽  
Growing Season ◽  
Zygotic Embryos ◽  
Embryo Abortion ◽  
Blue Spruce ◽  
Picea Pungens ◽  
Initiation Frequency ◽  
Propagation Methods

AbstractThe ‘triple-blue’ cultivar of blue spruce (Picea pungens Hoopsii) is notably recalcitrant towards the realm of traditional vegetative propagation methods. Its ability to naturally proliferate is limited by ovule and embryo abortion during the growing season, leading to low viable seed yield. In this study, we established a protocol using somatic embryogenesis (SE) as a means of propagating this popular ornamental cultivar. We collected cones from Hoopsii trees at seven different timepoints throughout the growing season (mid-June to late July in Ottawa (Plant Hardiness Zone 5A)). Female megagametophytes were harvested following each collection and immature zygotic embryos were plated onto induction media. Early somatic embryos began developing from the embryonic tissue (ET) three to five weeks following induction. The highest ET initiation frequency occurred from embryos collected June 20–July 10, suggesting that developmental stage of the embryo was a significant factor in SE induction. The conversion of mature somatic embryos into plantlets (emblings) was completed in eight–ten weeks at a rate of 92.8%. In this study, we demonstrate that in vitro somatic embryogenesis using our optimized protocol is a fast and prolific method for the mass propagation of Hoopsii blue spruce. This is the first report on the production of somatic Hoopsii emblings.

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