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 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 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.

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

HortScience ◽  
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.

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.

Somatic embryogenesis in tissue cultures of Podophyllum hexandrum

1990 ◽  
Vol 68 (3) ◽  
pp. 487-491 ◽  
Author(s):  
N. Arumugam ◽  
Sant S. Bhojwani
Keyword(s):  
Somatic Embryogenesis ◽  
Somatic Embryos ◽  
Basal Medium ◽  
Podophyllum Hexandrum ◽  
Zygotic Embryos ◽  
Tissue Cultures ◽  
Globular Embryos ◽  
Further Development ◽  
Sucrose Level

In vitro multiplication of Podophyllum hexandrum Royle (Podophyllaceae) via somatic embryogenesis is reported. The callus derived from zygotic embryos on Murashige and Skoog medium containing 2 μM BA and 0.5μM IAA differentiated globular embryos. On this medium the globular embryos continued to multiply but failed to mature. Further development of the embryos occurred if the sucrose level in the basal medium was raised to 6% or the medium was supplemented with 1–10 μM NAA. Light and temperatures higher than 25 °C suppressed embryogenesis. Embryogenic potential of the callus has been maintained for over 20 months through subcultures. The somatic embryos developed into plantlets on the basal medium. Key words: endangered species, podophyllotoxin, Podophyllum, somatic embryogenesis.

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.

In vivo verification of Cronartium ribicola propagated on tissue cultures of Pinus monticola

1970 ◽  
Vol 48 (7) ◽  
pp. 1429-1430 ◽  
Author(s):  
A. E. Harvey ◽  
J. L. Grasham
Keyword(s):  
Infected Tissue ◽  
Cronartium Ribicola ◽  
Tissue Cultures ◽  
Pinus Monticola ◽  
White Pine ◽  
Rust Infection ◽  
Western White Pine ◽  
Host Tissues

Inoculations of western white pine seedling stems with rust-infected tissue cultures produced one successful rust infection after 5 months. The infection was typical of this rust (Cronartium ribicola J. C. Fisch. ex Rabenh.) and the presence of haustoria was confirmed. Infected cortex tissue from this seedling was used to reestablish the isolate on host tissues grown in vitro.

scholarly journals Somatic embryogenesis from zygotic embryos of Euterpe oleracea Mart.

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 ◽  
...  
Keyword(s):  
Somatic Embryogenesis ◽  
Somatic Embryos ◽  
Zygotic Embryos ◽  
Ms Medium ◽  
Euterpe Oleracea ◽  
Mineral Salts ◽  
Vegetable Material ◽  
Morphogenetic Responses ◽  
Euterpe Oleracea Mart

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.

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.

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