scholarly journals Influence of Abscisic Acid and Sucrose on Somatic Embryogenesis in CactusCopiapoa tenuissimaRitt. formamostruosa

2013 ◽  
Vol 2013 ◽  
pp. 1-7 ◽  
Author(s):  
J. Lema-Rumińska ◽  
K. Goncerzewicz ◽  
M. Gabriel
Keyword(s):  
Somatic Embryogenesis ◽  
Abscisic Acid ◽  
Somatic Embryos ◽  
Sucrose Concentration ◽  
Turgor Pressure ◽  
Direct Somatic Embryogenesis ◽  
Fresh Weight ◽  
High Concentration ◽  
Globular Stage ◽  
Indirect Somatic Embryogenesis

Having produced the embryos of cactusCopiapoa tenuissimaRitt. formamonstruosaat the globular stage and callus, we investigated the effect of abscisic acid (ABA) in the following concentrations: 0, 0.1, 1, 10, and 100 μM on successive stages of direct (DSE) and indirect somatic embryogenesis (ISE). In the indirect somatic embryogenesis process we also investigated a combined effect of ABA (0, 0.1, 1 μM) and sucrose (1, 3, 5%). The results showed that a low concentration of ABA (0-1 μM) stimulates the elongation of embryos at the globular stage and the number of correct embryos in direct somatic embryogenesis, while a high ABA concentration (10–100 μM) results in growth inhibition and turgor pressure loss of somatic embryos. The indirect somatic embryogenesis study in this cactus suggests that lower ABA concentrations enhance the increase in calli fresh weight, while a high concentration of 10 μM ABA or more changes calli color and decreases its proliferation rate. However, in the case of indirect somatic embryogenesis, ABA had no effect on the number of somatic embryos and their maturation. Nevertheless, we found a positive effect of sucrose concentration for both the number of somatic embryos and the increase in calli fresh weight.

scholarly journals Induction of Somatic Embryogenesis in Vulnerable Medicinal Plant Hedychium spicatum Buch-Ham ex Smith

2014 ◽  
Vol 23 (2) ◽  
pp. 147-155 ◽  
Author(s):  
Dinesh Giri ◽  
Sushma Tamta
Keyword(s):  
Somatic Embryogenesis ◽  
Medicinal Plant ◽  
Somatic Embryos ◽  
Synthetic Seeds ◽  
Ex Situ ◽  
Secondary Embryogenesis ◽  
Zygotic Embryos ◽  
High Concentration ◽  
Cotyledon Explants ◽  
Short Period

This protocol has been developed for somatic embryogenesis in Hedychium spicatum. Simultaneously, a method has also been developed for the production of synthetic seeds by using somatic embryos. Direct somatic embryos were developed on cotyledon explants of zygotic embryos on MS supplemented with high concentration of NAA (20.0 µM). Induction of secondary embryogenesis was best in 2,4-D supplemented medium fortified with activated charcoal. Germination of somatic embryos was enhanced by using GA3. Besides this, round and semi-hard beads of somatic embryos (synthetic seeds) could be produced by using 2% Na-alginate and 100 mM calcium chloride and more than 30% germination of synthetic seeds was achieved in MS. Well acclimated plants produced via somatic embryogenesis and/or synthetic seeds were transferred to field where more than 60% survived. This simple study enabled us to obtain a number of plantlets throughout the year each cycle requiring a short period of time. Besides propagation, this study provided an ex situ method for conservation of this vulnerable Himalayan species.D. O. I.http://dx.doi.org/10.3329/ptcb.v23i2.17506Plant Tissue Cult. & Biotech. 23(2): 147-155, 2013  (December)

scholarly journals High-frequency Somatic Embryogenesis from Quiescent Seed Cotyledons of Cucumis melo Cultivars

1993 ◽  
Vol 118 (3) ◽  
pp. 425-432 ◽  
Author(s):  
D.J. Gray ◽  
D.W. McColley ◽  
Michael E. Compton
Keyword(s):  
Somatic Embryogenesis ◽  
Embryo Development ◽  
High Frequency ◽  
Somatic Embryos ◽  
Cucumis Melo ◽  
Sucrose Concentration ◽  
Male Sterile ◽  
Embryo Induction ◽  
Initial Culture ◽  
Dichlorophenoxy Acetic Acid

A protocol for high-frequency somatic embryogenesis in Cucumis melo L. was developed using `Male Sterile A147 as a model cultivar. Basal halves of quiescent seed cotyledons were cultured on embryo induction (EI) medium containing concentration ranges of the auxin 2,4-D and the cytokinins BA, Bin, TDZ, or 2iP before transfer to embryo development (ED) medium. Medium with 2,4-D at 5 mg·liter-1 and TDZ at 0.1 mg·liter-1 was superior, with 49% of explants responding and an average of 3.3 somatic embryos per explant (6.8 somatic embryos per responding explant). More explants produced embryos when incubated on EI medium for 1 or 2 weeks (30% and 33%) than for 3 or 4 weeks or with no induction. However, 2 weeks was 2.9 times better than 1 week in terms of number of embryos per explant. One week of initial culture in darkness, followed by a 16 hour light/8 hour dark photoperiod, produced more responding explants (26%) than two or more weeks in darkness or no dark period at all; but 1 and 2 weeks of darkness resulted in a similar number of embryos per explant (2.1 and 2.8). Sucrose concentration in EI and ED media had a highly significant effect on embryo induction and development. EI medium with 3% sucrose resulted in more embryogenic explants than EI medium with 1.5% or 6% sucrose. However, treatments with 3% sucrose in EI medium and 3% or 6% sucrose in ED medium produced significantly more embryos per explant (8.5 and 11.9) than other treatments. Treatments did not affect embryo induction directly and regeneration per se but, instead, frequency and efficiency of somatic embryo development. The optimal treatments were tested with 51 other commercial varieties. All varieties underwent somatic embryogenesis, exhibiting a response of 5% to 100% explant response and 0.1-20.2 embryos per explant. Chemical names used: N-(phenylmethyl)-lH-purin-6-amine (benzyladenine or BA); N-(2-furanylmethyl)-lH-purin-6-amine (kinetin or BIN); N-phenyl-N'-1,2,3-thiadiazol-5-ylurea (thidiazuron or TDZ); N-(3-methyl-2-butenyl)-lH-purin-6-amine (2iP); (2,4-dichlorophenoxy) acetic acid (2,4-D).

scholarly journals NAA at a high concentration promotes efficient plant regeneration via direct somatic embryogenesis and SE-mediated transformation system in Ranunculus sceleratus

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Ke-dong Xu ◽  
Wei Wang ◽  
De-shui Yu ◽  
Xiao-li Li ◽  
Jia-min Chen ◽  
...  
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Marker Gene ◽  
Direct Somatic Embryogenesis ◽  
The Novel ◽  
Transformation System ◽  
High Concentration ◽  
High Frequencies ◽  
Classical Pattern ◽  
Flower Organs

AbstractThe novel methods for efficient plant regeneration via direct somatic embryogenesis (SE) and SE-mediated transformation system under high concentration of NAA in Ranunculus sceleratus were established. On MS media containing a high concentration of NAA (10.0 mg/L) in the dark, all inoculated explants (root, stem and leaf) formed somatic embryos at high frequencies, respectively, 66.03, 126.47 and 213.63 embryoids per explant, and 100% of the embryoids developed into plantlets on 1/2 MS rooting media. Morphological and histological analyses revealed that SE in R. sceleratus followed a classical pattern. All inoculated explants can be used as receptors for genetic transformation in R. sceleratus, through direct SE-mediated method after Agrobacterium infection. RcLEC1-B, as a marker gene, changed the number and morphology of flower organs and the development of cuticle in R. sceleratus, which indicated that the efficient transgenic system of R. sceleratus was established. To our knowledge, this is the first observation that both direct SE and transgenic transformation system, via induction of a single plant growth regulator, have been successfully constructed in R. sceleratus.

Regeneration of Robiniapseudoacacia via somatic embryogenesis

1989 ◽  
Vol 19 (2) ◽  
pp. 285-288 ◽  
Author(s):  
S. A. Merkle ◽  
A. T. Wiecko
Keyword(s):  
Somatic Embryogenesis ◽  
Somatic Embryos ◽  
Basal Medium ◽  
Direct Somatic Embryogenesis ◽  
Secondary Embryogenesis ◽  
Dichlorophenoxyacetic Acid ◽  
Entire Study ◽  
Fruit Maturity ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Free Media

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.

Propagation of interior spruce by somatic embryogenesis

1990 ◽  
Vol 20 (11) ◽  
pp. 1759-1765 ◽  
Author(s):  
F. B. Webster ◽  
D. R. Roberts ◽  
S. M. McInnis ◽  
B. C. S. Sutton
Keyword(s):  
Somatic Embryogenesis ◽  
Abscisic Acid ◽  
Relative Humidity ◽  
Somatic Embryos ◽  
Large Scale ◽  
Final Height ◽  
High Relative Humidity ◽  
Mature Embryos ◽  
Interior Spruce ◽  
General Production

To apply somatic embryogenesis to clonal propagation of forest species, the technique must be applicable to a broad range of genotypes and allow efficient regeneration of phenotypically normal plants. Seventy-one lines (genotypes) of embryogenic cultures from six open-pollinated families were obtained by culturing immature embryos of interior spruce. Interior spruce represents a mixture of two closely related species, Piceaglauca (Moench) Voss and Piceaengelmannii Parry, from the interior of British Columbia where they hydridize with one another. The abscisic acid dependent developmental profile (the proportion of rooty embryos, shooty embryos, precociously germinating embryos, and mature embryos over a range of abscisic acid concentrations) differed among genotypes, but in general, production of mature somatic embryos was highest at 40 and 60 μM abscisic acid. Treatment of mature embryos with a high relative humidity treatment resulted in partial drying of the embryos and upon rehydration, markedly enhanced germination of the eight genotypes tested. Within 1 week of being placed under germination conditions, somatic embryos treated with the high relative humidity treatment showed 80–100% germination for 12 of the genotypes, and most genotypes had germination rates of greater than 40%. Survival of "emblings" (germinants from somatic embryos) following transfer to soil, acclimatization, and first season's growth in the nursery was 80% or greater for most genotypes. Over 1200 emblings were tested for nursery performance, representing the first large-scale evaluation of conifer somatic embryos under exvitro conditions. Growth rates, final height, shoot and root morphology, and frost hardiness were similar for emblings and seedlings following the first growing season. These results indicate that somatic embryogenesis can be used for the production of planting stock for a range of interior spruce genotypes.

scholarly journals Embriogenesis somatik langsung dan regenerasi planlet kopi arabika (Coffea arabica) dari berbagai eksplan Direct somatic embryogenesis and regeneration of arabica coffee plantlets (Coffea arabica) from different explants

2016 ◽  
Vol 71 (2) ◽  
Author(s):  
Fetrina OKTAVIA ◽  
. SWANTO ◽  
Asmini BUDIANI
Keyword(s):  
Somatic Embryogenesis ◽  
Growth Regulator ◽  
Somatic Embryos ◽  
Coffea Arabica ◽  
Culture Technique ◽  
Direct Somatic Embryogenesis ◽  
Root Explants ◽  
Soil Medium ◽  
Maturation Medium ◽  
Primary Somatic Embryos

SummaryTissue culture technique for arabica coffeefaces some problems, mainly in plantletsregeneration from cultured explants. Theobjectives of this experiment were to examine theeffect 2,4-D and 2-ip combinations on somaticembryogenesis and regeneration of arabicacoffee from several different explants. Basalmedium used in this experiment was MS mediumwith ½ concentration of macro and micro salts.Experiment to induce primary somatic embryos(SE) was arranged in factorial randomizedcomplete design with 10 repeats. The first factorwas the type of explants, leaf, epicotyl, hipocotyland root explants. The second factor was plantgrowth regulator i.e. combination of 1  M 2,4-Dwith 5, 10, 15, 20  M and combination of 5  M2,4-D with 5, 10, 15 and 20  M 2-ip. To multiplySE, secondary SE was induced from primary SEon medium containing combination of 0.6  MIAA and 13.3; 17.8 and 22.2  M BAP.Cotyledonary SE were germinated on mediacontaining GA 3 (0, 5, 10 and 15  M), and thenregenerated on medium free of growth regulator.Plantlets with 4-5 leaf pairs were transfered intothe soil medium for acclimatization. The resultsshow that primary SE can be induced from allexplants with the highest frequency on mediumcontaining 1  M 2,4-D and 15  M 2-ip.Induction of primary SE, in leaf explant wasmore effective than other explants. Mediumcontaining 0.6  M IAA and 22.2  M BAP gavethe highest percentage of SE multiplication i.e.52.6% with average SE number of 6.25. Plantletsregeneration can be conducted by culturing SEon maturation medium free of growth regulatorfor one month followed by germinating onmedium containing GA 3 , and then culturing onmedium free of growth regulator again. Thehighest percentage of germinated embryos wasobtained after three weeks and six weekscultured in the medium containing 5  M GA 3 , i.e49% and 90.15 respectively. From total plantletsobtained, 75% of them were normal. Sixtypercents of the young plants grew well in thegreenhouse.RingkasanTeknik kultur jaringan tanaman kopi arabikamasih menghadapi beberapa kendala terutamapada tingkat regenerasi planlet dari eksplan yangdikulturkan. Penelitian ini bertujuan untukmengetahui pengaruh kombinasi 2,4-D dan 2-ipterhadap embriogenesis somatik dan regenerasikopi arabika dari berbagai eksplan. Media dasaryang digunakan adalah medium MS ½konsentrasi garam makro dan mikro. Percobaaninduksi embrio somatik (ES) primer disusunmenurut rancangan acak lengkap faktorial dengan10 ulangan. Faktor pertama adalah jenis eksplan,erdiri atas daun, epikotil, hipokotil dan akar invitro. Faktor kedua adalah zat pengatur tumbuh,yaitu kombinasi 1 M 2,4-D dengan 5, 10, 15dan 20M 2-ip, serta kombinasi 5 M 2,4-Ddengan 5, 10, 15 dan 20 M 2-ip. Untuk mem-perbanyak jumlah ES yang didapatkan, dilakukaninduksi ES sekunder dari ES primer pada mediumyang mengandung kombinasi 0,6 M IAA dan13,3; 17,8 dan 22,2 M BAP. ES fase kotiledonkemudian dikecambahkan pada medium yangmengandung GA 3 (0, 5, 10 dan 15 M) danselanjutnya diregenerasikan pada medium tanpazat pengatur tumbuh. Planlet yang mempunyai4-5 pasang daun dipindahkan ke medium tanahuntuk aklimatisasi. Hasil yang diperolehmenunjukkan bahwa ES primer dapat diinduksipada semua eksplan yang digunakan denganfrekuensi tertinggi pada medium yang me-ngandung 1 M 2,4-D dan 15 M 2-ip. InduksiES primer pada eksplan daun lebih efektifdibandingkan eksplan lainnya. Untuk per-banyakan ES, medium yang mengandung IAA0,6 M dan BAP 22,2 M memberikanpersentase tertinggi pembentukan ES sekunderyaitu 52,6% dengan rata-rata jumlah ES 6,25.Regenerasi planlet dapat dilakukan denganmengkulturkan ES pada medium maturasi tanpazat pengatur tumbuh selama satu bulan, kemudiandikecambahkan dalam medium yang mengan-dung GA 3 , dan selanjutnya dipindah ke mediumtanpa zat pengatur tumbuh kembali.Perkecambahan ES tertinggi diperoleh padamedium dengan penambahan GA 3 5 M yaitu40,9% setelah tiga minggu dan 90,1% setelahenam minggu. Dari total planlet diperoleh 75%planlet normal. Hasil aklimatisasi menunjukkanbahwa 60% bibit mampu bertahan di rumah kaca.

A leucine-rich repeat containing receptor-like kinase marks somatic plant cells competent to form embryos

Development ◽  
1997 ◽  
Vol 124 (10) ◽  
pp. 2049-2062 ◽  
Author(s):  
E.D. Schmidt ◽  
F. Guzzo ◽  
M.A. Toonen ◽  
S.C. de Vries
Keyword(s):  
Somatic Embryogenesis ◽  
Somatic Embryos ◽  
Zygotic Embryo ◽  
Single Cells ◽  
Leucine Rich Repeat ◽  
Embryogenic Cells ◽  
Hypocotyl Explants ◽  
Globular Stage ◽  
Receptor Like Kinase ◽  
Small Subpopulation

The first somatic single cells of carrot hypocotyl explants having the competence to form embryos in the presence of 2,4-dichlorophenoxyacetic acid (2,4-D) were identified using semi-automatic cell tracking. These competent cells are present as a small subpopulation of enlarged and vacuolated cells derived from cytoplasm-rich and rapidly proliferating non-embryogenic cells that originate from the provascular elements of the hypocotyl. A search for marker genes to monitor the transition of somatic into competent and embryogenic cells in established suspension cell cultures resulted in the identification of a gene transiently expressed in a small subpopulation of the same enlarged single cells that are formed during the initiation of the embryogenic cultures from hypocotyl explants. The predicted amino acid sequence and in vitro kinase assays show that this gene encodes a leucine-rich repeat containing receptor-like kinase protein, designated Somatic Embryogenesis Receptor-like Kinase (SERK). Somatic embryos formed from cells expressing a SERK promoter-luciferase reporter gene. During somatic embryogenesis, SERK expression ceased after the globular stage. In plants, SERK mRNA could only be detected transiently in the zygotic embryo up to the early globular stage but not in unpollinated flowers nor in any other plant tissue. These results suggest that somatic cells competent to form embryos and early globular somatic embryos share a highly specific signal transduction chain with the zygotic embryo from shortly after fertilization to the early globular embryo.

scholarly journals The Fate of Integrated Ri T-DNA rol Genes during Regeneration via Somatic Embryogenesis in Tylophora indica

2015 ◽  
Vol 2015 ◽  
pp. 1-16 ◽  
Author(s):  
Dipasree Roychowdhury ◽  
Binay Chaubey ◽  
Sumita Jha
Keyword(s):  
Somatic Embryogenesis ◽  
Embryogenic Callus ◽  
Somatic Embryos ◽  
Growth Morphology ◽  
Tylophora Indica ◽  
Transformed Plants ◽  
Indirect Somatic Embryogenesis ◽  
Spontaneous Regeneration ◽  
Callus Lines

The fate of integrated Ri T-DNA rol genes during regeneration via indirect somatic embryogenesis and stability of its effect on morphology and tylophorine content of Ri-transformed plants have been studied in Tylophora indica. Integration and expression of Ri T-DNA genes in transformed embryogenic callus lines derived from transformed root lines, 300 Ri-transformed somatic embryos, and 23 Ri-transformed plant lines were analysed. Fifty root lines studied showed integration and expression of four rol genes of TL-DNA. Spontaneous regeneration via indirect somatic embryogenesis was obtained from root lines that were TL+/TR−. Stable integration and expression of rol genes were observed in root lines, embryogenic callus lines, and the spontaneously induced somatic embryos. Nineteen out of the 23 Ri-transformed plant lines and their clones showed phenotypic and genetic stability over the period of 3 years. Four Ri-transformed plants were morphologically similar to nontransformed plants but showed variation with the integration and expression of the rolA gene and absence of other rol genes. Variant Ri-transformed plant line A428#1-V showed highest tylophorine content (2.93±0.03 mg gDW−1) among plant lines studied. The effects of T-DNA genes on growth, morphology, and tylophorine content of the Ri-transformed plants were stable in the long term culture.

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