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 In vitro regeneration and somatic embryogenesis in citrus

2015 ◽  
Vol 24 (2) ◽  
pp. 247-262 ◽  
Author(s):  
El Sawy A Mohamed ◽  
Amina Gomaa ◽  
Nancy Danial
Keyword(s):  
Somatic Embryogenesis ◽  
Genetic Stability ◽  
Somatic Embryos ◽  
Rapd Analysis ◽  
In Vitro Regeneration ◽  
Rt Pcr ◽  
Mother Plants

Better results were obtained when stigma explants of variegated lemon and citron were used. After ten months, somatic embryos developed into plantlets at a frequency ranged from 13.3 for lime to 66.7% for lemon. Virus presence was tested by ELISA and RT?PCR. The results indicated that the plantlets regenerated through somatic embryogenesis are CTV?free. RAPD analysis was used to asses the genetic stability of plantlets as compared to the mother plants. The results indicated that most plantlets belong to the respective mother plants and the polymorphism percentage was genotype and explant?dependant.Plant Tissue Cult. & Biotech. 24(2): 247-262, 2014 (December

scholarly journals Recurrent somatic embryogenesis in long-term cultures of Gentiana lutea L. as a source for synthetic seed production for medium-term preservation

2012 ◽  
Vol 64 (2) ◽  
pp. 809-817 ◽  
Author(s):  
Irina Holobiuc ◽  
R. Catana
Keyword(s):  
Somatic Embryogenesis ◽  
Seed Production ◽  
Somatic Embryos ◽  
Plant Material ◽  
Synthetic Seed ◽  
Sugar Alcohols ◽  
Medium Term ◽  
Gentiana Lutea

Our aim was to establish an efficient and reproducible system for producing synthetic seeds from recurrent somatic embryogenesis in long-term cultures of Gentiana lutea L. This species is a vulnerable medicinal plant, protected both at the national and international levels, and is included in different Red Lists and Books. In vitro culture, as an alternative to classical methods of preservation, allows for the cyclic multiplication of plant material and short-, medium- and long-term preservation of tissue collections. Biotechnological approaches allow for maintenance of the plant material in a confined space and protection against biotic and abiotic factors. Somatic embryogenesis (SE) is the most efficient way to regenerate plants, ensuring material for preservation and fundamental research. In our experiment, recurrent somatic embryogenesis was developed in long-term cultures in the presence of sugar alcohols (mannitol, sorbitol) and in the absence of growth factors. This process proceeded at a high rate, with adventive somatic embryos being generated in a continuous process, followed by maturation, germination and development into plants. To follow the somatic embryogenesis process, histological samples were made. We used these embryogenic cultures for synthetic seed production and medium-term conservation. The viability of somatic embryos after moderate osmotic stress treatment was tested using TTC. Our methodology relied on the induction of somatic embryogenesis in the presence of auxins in the first cycle of in vitro cultures, long-term high embryogenic culture maintenance in the presence of sugar alcohols and synthetic seed production.

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.

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 Protocol development for somatic embryogenesis, SSR markers and genetic modification of Stipagrostis pennata (Trin.) De Winter

Plant Methods ◽  
2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Masoumeh Asadi-Aghbolaghi ◽  
Beata Dedicova ◽  
Sonali Sachi Ranade ◽  
Kim-Cuong Le ◽  
Farzad Sharifzadeh ◽  
...  
Keyword(s):  
Somatic Embryogenesis ◽  
Somaclonal Variation ◽  
Ssr Markers ◽  
Genetic Stability ◽  
Somatic Embryos ◽  
Large Scale ◽  
Scale Production ◽  
Ms Medium ◽  
Important Species ◽  
Regenerated Plants

Abstract Background Stipagrostis pennata (Trin.) De Winter is an important species for fixing sand in shifting and semi-fixed sandy lands, for grazing, and potentially as a source of lignocellulose fibres for pulp and paper industry. The seeds have low viability, which limits uses for revegetation. Somatic embryogenesis offers an alternative method for obtaining large numbers of plants from limited seed sources. Results A protocol for plant regeneration from somatic embryos of S. pennata was developed. Somatic embryogenesis was induced on Murashige & Skoog (MS) medium supplemented with 3 mg·L–1 2,4-D subsequently shoots were induced on MS medium and supplemented with 5 mg·L–1 zeatin riboside. The highest shoots induction was obtained when embryogenic callus derived from mature embryos (96%) in combination with MS filter-sterilized medium was used from Khuzestan location. The genetic stability of regenerated plants was analysed using ten simple sequence repeats (SSR) markers from S. pennata which showed no somaclonal variation in regenerated plants from somatic embryos of S. pennata. The regenerated plants of S. pennata showed genetic stability without any somaclonal variation for the four pairs of primers that gave the expected amplicon sizes. This data seems very reliable as three of the PCR products belonged to the coding region of the genome. Furthermore, stable expression of GUS was obtained after Agrobacterium-mediated transformation using a super binary vector carried by a bacterial strain LBA4404. Conclusion To our knowledge, the current work is the first attempt to develop an in vitro protocol for somatic embryogenesis including the SSR marker analyses of regenerated plants, and Agrobacterium-mediated transformation of S. pennata that can be used for its large-scale production for commercial purposes.

Somatic embryogenesis and plant regeneration from immature embryos of Carica papaya × Carica cauliflora cultured in vitro

1991 ◽  
Vol 69 (9) ◽  
pp. 1913-1918 ◽  
Author(s):  
M. H. Chen ◽  
C. C. Chen ◽  
D. N. Wang ◽  
F. C. Chen
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Somatic Embryos ◽  
Interspecific Hybrids ◽  
Carica Papaya ◽  
Immature Embryos ◽  
Embryogenic Cultures ◽  
Regenerated Plants ◽  
Hybrid Embryo

Somatic embryos were induced directly on immature embryos of Carica papaya × Carica cauliflora hybrids cultured on modified Murashige and Skoog's medium. When transferred to medium supplemented with abscisic acid, individual somatic embryos proliferated numerous daughter embryos through repeated embryogenesis. Light microscopic study of the repeatedly embryogenic cultures showed that daughter embryos arose from single superficial cells of parent embryos. Plant regeneration occurred following transfer of somatic embryos to medium devoid of plant growth regulators. Regenerated plants were intermediate between C. papaya and C. cauliflora in several morphological respects and showed isozyme patterns specific to both species as well as some new bands, indicating that they are indeed interspecific hybrids. Key words: Carica, interspecific hybrid, embryo culture, somatic embryogenesis.

Somatic Embryogenesis and In vitro Plant Regeneration in Vigna trilobata (L.) Verd. - A Potential Pasture Legume

1970 ◽  
Vol 19 (1) ◽  
pp. 89-99
Author(s):  
K. Choudhary ◽  
M. Singh ◽  
M. S. Rathore ◽  
N. S. Shekhawat
Keyword(s):  
Somatic Embryogenesis ◽  
Growth Regulators ◽  
Somatic Embryos ◽  
Basal Medium ◽  
Long Term Study ◽  
Continuous Application ◽  
First Time ◽  
Pasture Legume

This long term study demonstrates for the first time that it is possible to propagate embryogenic Vigna trilobata and to subsequently initiate the differentiation of embryos into complete plantlets. Initiation of callus was possible on 2,4-D. Somatic embryos differentiated on modified MS basal nutrient medium with 1.0 mg/l  of 2,4-D and 0.5 mg/l  of Kn. Sustained cell division resulted in globular and heart shape stages of somatic embryos. Transfer of embryos on to a fresh modified MS basal medium with 0.5 mg/l of Kn and 0.5 mg/l of GA3 helped them to attain maturation and germination. However, the propagation of cells, as well as the differentiation of embryos, were inhibited by a continuous application of these growth regulators. For this reason, a long period on medium lacking these growth regulators was necessary before the differentiation of embryos occurred again. The consequences for improving the propagation of embryogenic cultures in Vigna species are discussed. Key words: Pasture  legume, Vigna trilobata, Globular, Heart shape, somatic embryogenesis D.O.I. 10.3329/ptcb.v19i1.4990 Plant Tissue Cult. & Biotech. 19(1): 89-99, 2009 (June)

scholarly journals Telomere Length in Norway Spruce during Somatic Embryogenesis and Cryopreservation

Plants ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 416
Author(s):  
Tuija Aronen ◽  
Susanna Virta ◽  
Saila Varis
Keyword(s):  
Somatic Embryogenesis ◽  
Telomere Length ◽  
Norway Spruce ◽  
Genotypic Variation ◽  
Production Capacity ◽  
Stress Factors ◽  
Embryo Production ◽  
One Year ◽  
Plant Telomeres

Telomeres i.e., termini of the eukaryotic chromosomes protect chromosomes during DNA replication. Shortening of telomeres, either due to stress or ageing is related to replicative cellular senescence. There is little information on the effect of biotechnological methods, such as tissue culture via somatic embryogenesis (SE) or cryopreservation on plant telomeres, even if these techniques are widely applied. The aim of the present study was to examine telomeres of Norway spruce (Picea abies (L.) Karst.) during SE initiation, proliferation, embryo maturation, and cryopreservation to reveal potential ageing or stress-related effects that could explain variation observed at SE process. Altogether, 33 genotypes from 25 families were studied. SE initiation containing several stress factors cause telomere shortening in Norway spruce. Following initiation, the telomere length of the embryogenic tissues (ETs) and embryos produced remains unchanged up to one year of culture, with remarkable genotypic variation. Being prolonged in vitro culture can, however, shorten the telomeres and should be avoided. This is achieved by successful cryopreservation treatment preserving telomere length. Somatic embryo production capacity of the ETs was observed to vary a lot not only among the genotypes, but also from one timepoint to another. No connection between embryo production and telomere length was found, so this variation remains unexplained.

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

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