scholarly journals Differential success of somatic embryogenesis in random gene pool of Norway spruce

2008 ◽  
Vol 53 (No. 2) ◽  
pp. 74-87
Author(s):  
M. Mauleová ◽  
J. Vítámvás
Keyword(s):  
Somatic Embryogenesis ◽  
Norway Spruce ◽  
Somatic Embryos ◽  
Significant Variation ◽  
Open Pollination ◽  
Embryogenic Cultures ◽  
Peg 4000 ◽  
Partial Desiccation ◽  
Somatic Embryogenic ◽  
Mature Seeds

Somatic embryogenic cultures were established from proembryonal suspensor masses (PEMs) derived from mature seeds of Norway spruce. In this study we used more than 4,300 seeds of <i>Picea abies</i> from randomly collected commercial seed lot (originated from open-pollination). Most of the studies are focused on selected genotypes known for higher response to propagation protocols. As indicated in this study, there is a significant variation in success rate of somatic embryogenesis in randomly selected seed lot of Norway spruce. Nutrient GD (1 to 4), LP (1 to 5) media and different level of plant grow regulators (BA, NAA, kinetin and 2,4D) were used for initiation and proliferation of embryogenic cultures. Transfer of embryogenic callus onto medium containing abscisic acid stimulated development of early-established individual embryos. Media GD (5 and 6) and LP (9 to 11) supplemented with ABA (7.5; 20; 38 &mu;M) and PEG 4000 (2%), were used for stadium of maturation. Conversion of somatic embryos to plantlets was stimulated by partial desiccation treatment (HRH-treatment) and by medium changes. On these media plantlets started to regenerate within three weeks.

Somatic embryogenesis in mature zygotic embryos of Picea likiangensis

Biologia ◽  
2010 ◽  
Vol 65 (5) ◽  
Author(s):  
Shaoyu Chen ◽  
Shanna Chen ◽  
Fang Chen ◽  
Tao Wu ◽  
Yinbin Wang ◽  
...  
Keyword(s):  
Somatic Embryogenesis ◽  
Somatic Embryos ◽  
Clonal Propagation ◽  
Zygotic Embryos ◽  
Dichlorophenoxyacetic Acid ◽  
Embryogenic Cultures ◽  
Peg 4000 ◽  
Half Strength ◽  
Basal Media ◽  
2,4 Dichlorophenoxyacetic Acid

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.

EFFECT OF 2,4-D AND NAA ON SOMATIC EMBRYOGENESIS FROM APICAL PORTION OF GROUNDNUT (ARACHIS HYPOGAEA L.)

2021 ◽  
Vol 1 (7) ◽  
pp. 119-124
Author(s):  
Muniappan V ◽  
Manivel P ◽  
Prabakaran V ◽  
Palanivel S ◽  
Parvathi S
Keyword(s):  
Somatic Embryogenesis ◽  
Somatic Embryo ◽  
Arachis Hypogaea ◽  
Somatic Embryos ◽  
Mature Embryo ◽  
Induction Medium ◽  
Embryogenic Cultures ◽  
Apical Portion ◽  
Arachis Hypogaea L ◽  
Somatic Embryo Induction

Somatic embryogenesis was carried out epicotyl portion of the mature embryo/apical portion. The somatic embryo induction medium containing 2,4-D or NAA (10.0 to 50.0 mg/l). Of the two concentrations tested 2,4-D (30.0mg/l) recorded the highest percentage of response followed by NAA (30.0mg/l). But the highest number of somatic embryo were recorded in 30.0mg/l of 2,4-D followed by NAA. The apical portion of the mature embryo formed direct embryos without any intervention of callus. The maximum percentage of embryogenic cultures were noticed in 30.0mg/l of 2,4-D followed by NAA at 30.0mg/l. for the differentiation of somatic embryos, the embryogenic masses were transferred to medium without any growth regulator. The maximum number of somatic embryos per culture was recorded in 30 mg/l of 2,4-D followed by 30.0 mg/l of NAA. Keywords: Arachis hypogaea L.,Somatic Embryogenesis, 2,4-D and NAA

scholarly journals Somaclonal Variation During Picea abies and P. omorika Somatic Embryogenesis and Cryopreservation

2017 ◽  
Vol 59 (1) ◽  
pp. 93-103 ◽  
Author(s):  
Teresa Hazubska-Przybył ◽  
Monika Dering
Keyword(s):  
Somatic Embryogenesis ◽  
Picea Abies ◽  
Somaclonal Variation ◽  
Genetic Stability ◽  
Somatic Embryos ◽  
Plant Material ◽  
Stress Factors ◽  
Embryogenic Cultures ◽  
Embryogenic Lines

AbstractEmbryogenic cultures of plants are exposed to various stress factors bothin vitroand during cryostorage. In order to safely include the plant material obtained by somatic embryogenesis in combination with cryopreservation for breeding programs, it is necessary to monitor its genetic stability. The aim of the present study was the assessment of somaclonal variation in plant material obtained from embryogenic cultures ofPicea abies(L.) Karst. andP. omorika(Pančić) Purk. maintainedin vitroor stored in liquid nitrogen by the pregrowth-dehydration method. The analysis of genetic conformity with using microsatellite markers was performed on cotyledonary somatic embryos (CSE), germinating somatic embryos (GSE) and somatic seedlings (SS), obtained from tissues maintainedin vitroor from recovered embryogenic tissues (ETc) and CSE obtained after cryopreservation. The analysis revealed changes in the DNA of somatic embryogenesis-derived plant material of bothPiceaspp. They were found in plant material from 8 out of 10 tested embryogenic lines ofP. abiesand in 10 out of 19 embryogenic lines ofP. omorikaafterin vitroculture. Changes were also detected in plant material obtained after cryopreservation. Somaclonal variation was observed in ETc and CSE ofP. omorikaand at ETv stage ofP. abies. However, most of the changes were induced at the stage of somatic embryogenesis initiation. These results confirm the need for monitoring the genetic stability of plants obtained by somatic embryogenesis and after cryopreservation for both spruce species.

scholarly journals Somatic Embryogenesis and Plant Regeneration in Ephedra foliata (Boiss.); a non Coniferous Gymnosperm

1970 ◽  
Vol 20 (2) ◽  
pp. 133-143 ◽  
Author(s):  
Manjul Dhiman ◽  
Vandana Sharma ◽  
Sushma Moitra
Keyword(s):  
Somatic Embryogenesis ◽  
Somatic Embryos ◽  
Root Formation ◽  
Coarse Sand ◽  
Garden Soil ◽  
Longitudinal Splitting ◽  
Ephedra Foliata ◽  
Best Responses ◽  
Mature Seeds

Embryos from mature seeds of Ephedra foliata were excised and used as explant after longitudinal splitting. Half embryos were cultured in vitro to assess their morphogenic potential. When 2 - 10 µM  2,4-D with 2 - 10 µM Kn was used in MS with 10% coconut milk callus, roots and somatic embryos were induced. Lower concentration of 2 and 5 µM 2,4-D with 2 to 15 µM Kn gave best responses in terms of percentage of cultures showing somatic embryos. greater degree of callusing was obtained in a combination of 8 - 10 µM 2,4-D with Kn, but frequency of somatic embryos was low. Substituting BAP for Kn in the medium containing 2,4-D showed almost the same response as that in 2,4-D and Kn. The percentage of cultures showing root formation was low as compared to Kn combinations. Somatic embryos developed, but with a lesser frequency. When different concentrations of NAA were added with Kn, only rooting and callusing occurred. On this medium, there was a total absence of somatic embryos. When somatic embryos were transferred to a hormone free medium, they grew within 10 - 15 days. The fully grown somatic embryos then germinated and developed into plantlets. Chromosomal count confirmed retention of ploidy level.  Plantlets from somatic embryos were transferred to pots containing sterilized mixture of coarse sand and garden soil in equal proportions. They survived well and formed new nodes and internodes after nearly one month.   Key words:  Ephedra, Somatic embryogenesis, Micropropagation, Ephedrine   D.O.I. 10.3329/ptcb.v20i2.6893   Plant Tissue Cult. & Biotech. 20(2): 133-143, 2010 (December)

Two waves of programmed cell death occur during formation and development of somatic embryos in the gymnosperm, Norway spruce

2000 ◽  
Vol 113 (24) ◽  
pp. 4399-4411 ◽  
Author(s):  
L.H. Filonova ◽  
P.V. Bozhkov ◽  
V.B. Brukhin ◽  
G. Daniel ◽  
B. Zhivotovsky ◽  
...  
Keyword(s):  
Cell Death ◽  
Somatic Embryogenesis ◽  
Programmed Cell Death ◽  
Norway Spruce ◽  
Somatic Embryos ◽  
Nuclear Dna ◽  
Second Phase ◽  
Embryo Suspensor ◽  
Large Central Vacuole ◽  
Suspensor Cells

In the animal life cycle, the earliest manifestations of programmed cell death (PCD) can already be seen during embryogenesis. The aim of this work was to determine if PCD is also involved in the elimination of certain cells during plant embryogenesis. We used a model system of Norway spruce somatic embryogenesis, which represents a multistep developmental pathway with two broad phases. The first phase is represented by proliferating proembryogenic masses (PEMs). The second phase encompasses development of somatic embryos, which arise from PEMs and proceed through the same sequence of stages as described for their zygotic counterparts. Here we demonstrate two successive waves of PCD, which are implicated in the transition from PEMs to somatic embryos and in correct embryonic pattern formation, respectively. The first wave of PCD is responsible for the degradation of PEMs when they give rise to somatic embryos. We show that PCD in PEM cells and embryo formation are closely interlinked processes, both stimulated upon withdrawal or partial depletion of auxins and cytokinins. The second wave of PCD eliminates terminally differentiated embryo-suspensor cells during early embryogeny. During the dismantling phase of PCD, PEM and embryo-suspensor cells exhibit progressive autolysis, resulting in the formation of a large central vacuole. Autolytic degradation of the cytoplasm is accompanied by lobing and budding-like segmentation of the nucleus. Nuclear DNA undergoes fragmentation into both large fragments of about 50 kb and multiples of approximately 180 bp. The tonoplast rupture is delayed until lysis of the cytoplasm and organelles, including the nucleus, is almost complete. The protoplasm then disappears, leaving a cellular corpse represented by only the cell wall. This pathway of cell dismantling suggests overlapping of apoptotic and autophagic types of PCD during somatic embryogenesis in Norway spruce.

scholarly journals Polyamines during somatic embryo development in Norway spruce (Picea abies [L.])

2009 ◽  
Vol 55 (No. 2) ◽  
pp. 75-80 ◽  
Author(s):  
J. Malá ◽  
M. Cvikrová ◽  
P. Máchová ◽  
O. Martincová
Keyword(s):  
Somatic Embryogenesis ◽  
Picea Abies ◽  
Norway Spruce ◽  
Somatic Embryos ◽  
Developmental Stages ◽  
Embryogenic Tissue ◽  
Growth Media ◽  
Mature Embryos ◽  
Proliferation Medium ◽  
Maturation Medium

Contents of free polyamines (putrescine, spermidine and spermine) were determined in different developmental stages of Norway spruce (<I>Picea abies</I> [L.] Karst.) somatic embryos by means of HPLC. Determinations were performed embryogenic tissue after 4 weeks of the growth on proliferation medium, after 2 and 5 weeks of the culturing on maturation medium, and 2 weeks after desiccation. Maturation of somatic embryos (after 5 weeks) was accompanied by increase of concentrations of putrescine (2.3 times) and spermidine (3.2 times). In comparison with above mentioned polyamines, spermine concentrations were significantly lower (4.3 times). Two weeks after desiccation, the concentrations of putrescine decreased 5.4 times and spermidine 2.2 times in comparison with mature embryos. To improve the efficiency of somatic embryogenesis of less responsive genotypes, the supplementation of growth media by polyamines is discussed.

scholarly journals Effect of Cryopreservation on Olive (Olea europaea L.) Plant Regeneration via Somatic Embryogenesis

Plants ◽  
2020 ◽  
Vol 10 (1) ◽  
pp. 34
Author(s):  
Fatiha Bradaï ◽  
Carolina Sánchez-Romero
Keyword(s):  
Somatic Embryogenesis ◽  
Somatic Embryos ◽  
Aluminum Foil ◽  
Regeneration Capacity ◽  
Fresh Weight ◽  
Negative Effects ◽  
Embryogenic Cultures ◽  
Embryogenic Line ◽  
Cryopreservation Protocol ◽  
Embryogenic Lines

Olive somatic embryos have been successfully cryopreserved using the droplet-vitrification method on aluminum foil strips. Although acceptable recovery rates have been obtained after rewarming, the influence of this cryopreservation protocol on the somatic embryogenesis process is unknown. To evaluate the effect of cryopreservation on olive somatic embryogenesis, the behavior of cultures established from cryopreserved somatic embryos was compared with that of control, non-cryopreserved cultures in the different phases of the somatic embryogenesis process. In order to analyze the influence of the genotype, this investigation was carried out in two independent lines. During the proliferation step, only the line T1 was affected by cryopreservation, with higher fresh weight increases. Although similar total embryos were produced per culture, freezing in liquid nitrogen significantly improved the maturation pattern in the line P5. Better germination results were also found in this embryogenic line. The genotype plays a key role, largely determining the effect of cryopreservation on olive somatic embryogenesis. A specific genotype-dependent response was found depending on the culture step. Variations observed could not be associated to differences in the embryogenic lines’ instability to maintain their morphogenic competence after cryopreservation. Embryogenic cultures established after rewarming retained their regeneration capacity, with no evident negative effects affecting their regeneration capacity.

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 Light Spectra during Somatic Embryogenesis of Norway Spruce—Impact on Growth, Embryo Productivity, and Embling Survival

Forests ◽  
2021 ◽  
Vol 12 (3) ◽  
pp. 301
Author(s):  
Saila Varis ◽  
Mikko Tikkinen ◽  
Sakari Välimäki ◽  
Tuija Aronen
Keyword(s):  
Somatic Embryogenesis ◽  
Norway Spruce ◽  
Somatic Embryos ◽  
Pulse Width Modulation ◽  
Production Costs ◽  
Embryogenic Tissue ◽  
Light Spectrum ◽  
Propagation Process ◽  
Embryo Survival ◽  
Light Spectra

For Norway spruce (Picea abies (L.) Karst.) somatic embryogenesis (SE) culture conditions throughout the propagation process affect the final result. Many critical phases can be identified, and all of them cumulatively increase the production costs of SE plants if they cannot be controlled. In order to determine the best lighting protocol for each SE step, Norway spruce embryogenic tissue (ET) was proliferated, and somatic embryos were matured under different light wavelengths, wavelength combinations, and in the dark. Overall, using low-intensity LED lights during proliferation or at the end of maturation had little effect on the growth of ET, embryo productivity, or embryo survival; on the other hand, major negative effects could not be seen. This is beneficial from a practical point of view, indicating no need for lighting or protection of SE cultures from light during their handling in these steps of the propagation process. When somatic embryos were germinated under different spectra, significant differences in embling shoot and root growth, as well as in the survival of the emblings, were found. The best treatment varied between trials, and the genotype of the SE culture was found to have a stronger effect than the light spectrum, indicating that various light spectra and also intensity adjusted using pulse width modulation (PWM) can be successfully applied to the SE germination phase in Norway spruce.

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