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  • Articles  (56)
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  • somatic embryogenesis
  • Springer  (56)
  • 1985-1989  (53)
  • 1955-1959  (3)
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  • Articles  (56)
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  • Springer  (56)
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  • 1
    Electronic Resource
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    Springer
    Plant cell reports 4 (1985), S. 285-288 
    ISSN: 1432-203X
    Keywords: Medicago sativa ; tissue culture ; regenerative ability ; somatic embryogenesis ; genetic variation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Nineteen accessions of diploid Medicago sativa L. belonging to the four subspecies sativa, caerula, falcata and xvaria were screened for their ability to produce somatic embryos on hypocotyl-derived callus. Two medium protocols were used in this study, a three-step sequence with exposure of the callus cultures to a high 2,4-D concentration and a two-step sequence without exposure to a high 2,4-D concentration. Considerable variation for callus proliferation was observed. In general, the diploid M. sativa accessions showed poor regenerability and it was not possible to correlate high regeneration frequencies with a particular germplasm source. It was, however, possible to identify regenerable genotypes in all four subspecies. One falcata accession produced somatic embryos on the callus induction media at high frequencies. This response was also obtained with a few genotypes from one xvaria accession. All regenerable plants were maintained as shoot cultures and were able to form somatic embryos on petiole-derived calli.
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  • 2
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    Plant cell reports 6 (1987), S. 231-234 
    ISSN: 1432-203X
    Keywords: Gossypium hirsutum ; somatic embryogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Tissue culture methods for improvement of cotton has lagged seriously compared to other major crops. A method for regeneration of cotton which includes a morphogenetically competent cell suspension was needed to facilitate selection of stress-resistant variants and gene manipulation. Preliminary screening of eight strains of Gossypium hirsutum L. for embryogenic potential resulted in the production of somatic embryos in all strains. Coker 312 was selected for use in the development of a model regeneration system for G. hirsutum. Calli were initiated from hypocotyl tissues of 3-day-old-seedlings. Globular embryos were present after six weeks in culture. Calli were subcultured to liquid suspension in growth regulator-free medium. After three to four weeks, suspensions were sieved to collect globular and heart stage embryos. Collected embryos developed further when plated onto semi-solid medium. To induce germination and plantlet growth, mature embryos were placed on sterile vermiculite saturated with medium. Upon development of roots and two true leaves, plantlets were potted in peat and sand, and hardened. Mature plants and progeny have been obtained with this procedure. A high percentage of infertile plants was observed among the regenerants.
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  • 3
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    Plant cell reports 8 (1989), S. 133-136 
    ISSN: 1432-203X
    Keywords: Cotton ; somatic embryogenesis ; cultivar
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Thirty eight cultivars, strains, and races ofGossypium were screened for somatic embryogenesis with the protocols developed as a model forG. hirsutum L. cv. Coker 312. Four classes of response were identified; high, moderate, low, and non-embryogenic. Four cultivars were further screened with 13 growth regulator regimes to determine if culture environment could change the classification or induce a higher level of response. The classification or level of response did not change. Screening of individual seedlings within a cultivar indicated that genotypic variation for embryogenesis existed. Highly embryogenic individuals were selected from cvs. Coker 312 and Paymaster 303 for use as germplasm sources for transfer of the embryogenic trait to other cultivars and genetic stocks. Only genetically responsive genotypes are amenable to the model developed for Coker 312.
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  • 4
    ISSN: 1432-203X
    Keywords: Oryza perennis Moench ; wild rice species ; somatic embryogenesis ; regeneration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Somatic embryogenesis in the wild rice species (Oryza perennis) was induced from cultured mature seeds and young inflorescences. Murashige and Skoog's (MS) medium supplemented with 2 mg/l 2,4-D and 0.2 mg/l BAP was used for induction of a compact, white nodular callus and somatic embryos. Plant regeneration occurred with the tranfer of the nodular callus to MS basal medium containing 0.5 mg/l IAA, 0.5 mg/l NAA, 4 mg/l BAP and 500 mg/l casein hydrolysate. The embryogenic nature of the callus from both explants was maintained over 10 subcultures for about 12 months. Plant regeneration with respect to the number of calli plated from the 6th to 10th passage varied from 80% to 60% for young inflorescence derived callus and from 75% to 69.8% for seed-derived callus.
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  • 5
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    Plant cell reports 7 (1988), S. 70-73 
    ISSN: 1432-203X
    Keywords: Albizia richardiana ; somatic embryogenesis ; organogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Hypocotyl explants of 1 and 10 mm lengths were excised from 12-day-old in vitro-grown seedlings of Albizia richardiana. The larger pieces, after 40 days of culture, developed shoots along with green calli on B5 + BAP (10−7–10−5M), while the smaller segments produced only green calli on B5+BAP (10−7–10−4M) medium. Some of the green calli turned morphogenic and started producing somatic embryos with the 2nd sub-culture and shoots from 7th sub-culture onwards. Calli retained the morphogenic potential even after repeated sub-culturing for over two years. The number of embryos in an embryogenic culture varied from 2 to 20 per callus mass of 5–6.5 cm3. Sucrose at the 2% level in MS medium was optimal for embryogenesis while 4% was optimal for shoot bud differentiation. Higher levels of sucrose (6–10%) caused browning of green calli and also inhibited differentiation into embryos and shoot buds. By selective sub-culturing of 0.1 cm3 pieces of embryogenic calli on MS+10−5M BAP, 46% of the cultures produced somatic embryos. The latter germinated into plantlets on Knop's medium.
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  • 6
    ISSN: 1432-203X
    Keywords: Populus ciliata ; somatic embryogenesis ; plant regeneration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Somatic embryogenesis and plantlet formation were obtained from callus and cell suspension cultures of 40-year- old Himalayan Poplar (Populus ciliata Wall ex Royle). Callus and cell suspensions were obtained by transfer of inoculum of semiorganized leaf cultures, which were maintained on Murashige and Skoog (MS) medium supplemented with benzylaminopurine (BAP), to MS with 2,4-dichlorophenoxyacetic acid (2,4-D). Reduction of 2,4-D concentration during subsequent subculture of cell suspensions resulted in the formation of embryoids. These embryoids developed further only after being transferred to agar-based MS medium supplemented with BAP and naphthalene acetic acid. Loss of embryogenic potential was observed in cell suspensions after 6 subcultures. However, callus cultures retained the embryogenic potential even after repeated subcultures for more than a year. Plantlets could be successfully hardened and grown in natural outdoor conditions.
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  • 7
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    Plant cell, tissue and organ culture 7 (1986), S. 257-261 
    ISSN: 1573-5044
    Keywords: Carica candamarcensis ; papaya ; somatic embryogenesis ; cell suspension cultures
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cell suspension cultures of Carica candamarcensis derived from hypocotyl calli were tested concerning their in vitro embryogenic capacity to improve asexual propagation rates in this species. Somatic embryos developed in culture from cells in suspension or from microcalli. Responses were affected by nutrient media and phytohormones used. Best results were obtained by growing the cells in suspension in Nitsch and Nitsch medium containing naphthaleneacetic acid and then plating them upon the same medium containing benzyladenine, or combinations of both hormones.
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  • 8
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    Plant cell, tissue and organ culture 9 (1987), S. 167-171 
    ISSN: 1573-5044
    Keywords: Vicia faba L. ; callus culture ; suspension culture ; somatic embryogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The paper describes a method of somatic embryo induction in callus and suspension cultures of Vicia faba L. Callus was induced from immature cotyledons (green maturity stage) of white-flowering horse bean lines cultured on L2 medium (Phillips and Collins 1979) supplemented with 1% sucrose, 0.7% agar and different concentrations of 2,4-dichlorophenoxyacetic acid. The medium with 2.5 μM 2,4-Dichlorophenoxyacetic acid was found optimum for embryogenic callus induction. Somatic embryos developed after transfer of the callus to media lower or zero 2,4-Dichlorophenoxyacetic acid and increased level of sucrose (2.5%). The release of somatic embryos from the callus was more apparent after transfer to liquid medium. There were various stages of somatic embryo development, i.e. globular, heart-shaped and torpedo ones.
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  • 9
    ISSN: 1573-5060
    Keywords: Oryza sativa L. cv. Taipei 309 ; rice ; cryogenic storage ; somatic embryogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The rates of recovery from cryogenic storage of suspension cultures of the Japonica rice cultivar Taipei 309, as determined by the reduction of triphenyl tetrazolium chloride and cell regrowth, were significantly influenced by the embryogenic potential of the non-frozen cultures.
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  • 10
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    Plant cell, tissue and organ culture 4 (1985), S. 249-259 
    ISSN: 1573-5044
    Keywords: Citrus sinensis Osb. ; nucellar callus ; protoplast ; somatic embryogenesis ; inhibitor(s) ; plant regeneration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Using ‘Trovita’ orange (Citrus sinensis Osb.) protoplasts isolated from 6-year-old nucellar callus, the effects of protoplast density and mannitol concentration on cell divisions and embryoid formation were examined. Somatic embryogenesis in nearly direct manner was observed only at a combination of low cell densities (∼4×104/ml) and low mannitol concentrations (∼0.4 M). Two alternatives to achieve high frequency embryogenesis (∼70%) were to either dilute the cells to lower densities, or to do serial transfers of cells to fresh medium. Orange protoplasts (cells) showed embryogenic potential, and repression of embryogenesis occurred when protoplasts were cultured at a high density and/or under high osmotic pressure.
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  • 11
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    Plant cell, tissue and organ culture 5 (1985), S. 151-162 
    ISSN: 1573-5044
    Keywords: Hordeum vulgare ; barley ; callus culture ; plant regeneration ; somatic embryogenesis ; organogenesis ; apical meristem
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Callus cultures were initiated from apical meristem explants of one to four-week-old aseptically-grown barley (Hordeum vulgare L. cv. Atlas 57) plants. Embryogenic callus and plants were produced in three separate experiments; the cultures have retained regenerative capacity for three years after initiation. Our results demonstrate that explants other than immature embryos are embryogenically competent in barley and that regeneration occurs by both somatic embryogenesis and organogenesis.
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  • 12
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    Plant cell, tissue and organ culture 5 (1986), S. 175-178 
    ISSN: 1573-5044
    Keywords: cereals ; somatic embryogenesis ; tissue culture ; Coix lacryma-jobi
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Callus was obtained from segments of immature inflorescence of Coix lacryma-jobi cultured on N6 medium containing 1–2 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) and 3–5% sucrose. Plantlets were regenerated when embryogenic calluses were transferred onto MS medium with 0.5 mg/l kinetin and 0.01 mg/l naphthaleneacetic acid (NAA). Regenerated plants had the diploid chromosome number (2n=20).
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  • 13
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    Hydrobiologia 132 (1986), S. 195-200 
    ISSN: 1573-5117
    Keywords: regeneration ; morphogenesis ; asexual reproduction ; somatic embryogenesis ; hypermorphosis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In reviewing recent research published in Russian on regeneration and asexual reproduction, the following morphogenetic processes in the planarian Dugesia tigrina are considered: 1) regeneration of lost parts of the body; 2) regeneration of the whole worm from fragments of the body, either by normal regeneration when the inital polarity of the fragment is retained or by somatic embryogenesis when one or more new axes of polarity arise; 3) somatic embryogenesis, or development of individuals from somatic cells; 4) hypermorphosis, or the presence of more than the usual number of organs or body parts, a process that can be interpreted in terms of somatic embryogenesis; and 5) asexual reproduction. Some morphological, biochemical, and physiological studies of the ‘division zone’ in D. tigrina demonstrate peculiarities of a local breakdown of integrative functions, a breakdown which in turn causes division of the individual to take place at this zone; timing of division is controlled by the organism as an integrated whole.
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  • 14
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    Plant cell, tissue and organ culture 10 (1987), S. 11-19 
    ISSN: 1573-5044
    Keywords: amino acids ; ammonium ; somatic embryogenesis ; tissue culture ; Medicago sativa
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effect of exogenously supplied reduced nitrogen and sucrose on high-frequency somatic embryogenesis in petiole-derived tissue cultures of a diploid and a tetraploid regenerable clone of Medicago sativa ssp. falcata was investigated. There was an absolute requirement for ammonium during embryo induction and differentiation, with 5mM being the optimum for induction and 10–20 mM the optimum for differentiation of somatic embryos. Exogenous amino acids were not essential for differentiation and often even inhibitory, except 1 or 2 g/l casein hydrolysate or 4.4 mM glutamine with 3.1 mM proline which, under certain conditions, resulted in increases of 20–30% in the number of embryos obtained. High and low sucrose concentrations inhibited somatic embryogenesis and there was no reason to deviate from the 3% (0.088 M) sucrose level commonly used in plant tissue culture media. Selected clones from three M. sativa cultivars showed a response similar to the highly regenerable ssp. falcata clone F1.1.
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  • 15
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    Plant cell, tissue and organ culture 10 (1987), S. 31-38 
    ISSN: 1573-5044
    Keywords: Old World bluestem grasses ; tissue culture ; apomixis ; somatic embryogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Explants from immature inflorescences of four genotypes of Old World bluestem grasses, (Bothriochloa spp.), produced callus tissue on Linsmaier and Skoog (RM) and 1/2 Murashige and Skoog (1/2 MS) media containing high levels of growth regulators. Callus masses were composed of two distinct tissue types, one a compact, white, embryogenic portion (E calli), the other soft, translucent, gelatinous and nonembryogenic (NE calli). When transferred to medium with a reduced level of 2,4-D, and/or supplemented with zeatin, E callus underwent further organization culminating in shoot production. Light and scanning electron microscopy confirmed the embryogenic pathway of differentiation. Genotype significantly affected callus induction frequency and the number of plants regenerated. The RM medium induced more explants to initiate callus compared to the 1/2 MS medium. Age of the inflorescence explant, as indicated by size, was critical for callus induction. Inflorescences with racemes ≤8 mm in length were superior to older ones. Five-hundred-twenty-two plantlets were regenerated and grown to maturity.
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  • 16
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    Plant cell, tissue and organ culture 10 (1987), S. 101-113 
    ISSN: 1573-5044
    Keywords: Wheat ; callus formation ; somatic embryogenesis ; 3,6-dichloro-o-anisic acid ; 2,4-dichlorophenoxyacetic acid ; 6-furfurylaminopurine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Somatic embryo (embryoid) formation from immature-embryo-derived calli was quantified in replicated experiments involving 10 Triticum aestivum L. genotypes. Several published media formulations, which had previously been optimized for wheat tissue culture, were tested for each genotype. Embryos from each plant were randomly assigned to each medium. Percentage precocious germination of immature embryos and mean percentage scutellar callus per explant were recorded. Embryoids per callus were determined by microscopic examination at 28 and 56 days. There were highly significant differences among genotypes, media, and individual plants from which explants were taken. A medium based on double the Murashige and Skoog (MS) inorganic salt concentration was significantly better than other media. Inclusion of all MS vitamins appeared essential for optimal response. Two genotypes were tested in a second experiment where both 3,6-dichloro-o-anisic acid (9.05 μM) and 6-furfurylaminopurine (0.46 μM) were substituted for 2,4-dichlorophenoxyacetic acid (4.52 μM) in either double or normal MS medium. This substitution significantly increased embryoid formation at 28 days. Additions of either 6-furfurylaminopurine or coconut water increased precocious germination of both embryo explants and embryoids.
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  • 17
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    Plant cell, tissue and organ culture 10 (1987), S. 149-156 
    ISSN: 1573-5044
    Keywords: Bactris gasipaes H.B.K. ; somatic embryogenesis ; organogenesis picloram
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Somatic embryogenesis in pejibaye or peach palm (Bactris gasipaes H.B.K.) was induced from callus derived from in vitro cultured shoot tips of young field-grown plants using a modified Murashige and Skoog medium supplemented with 5 mg L−1 of N6-benzyladenine (BA) and 0.06 mg L−1 of picloram for three months in the dark; this was followed by an additional three months with the same medium and incubation conditions, but using 0.03 mg L−1 of picloram. The cultures were then transferred to light on a medium without hormones. This led to the formation of morphogenic callus, in which somatic embryos, as well as shoot primordia, and finally complete plantlets were formed. These plantlets continued to grow on transfer to the greenhouse.
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  • 18
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    Plant cell, tissue and organ culture 10 (1987), S. 197-208 
    ISSN: 1573-5044
    Keywords: soybean ; somatic embryogenesis ; hormones ; tissue culture ; embryo morphology ; plant regeneration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Somatic embryos were induced in cultures of immature soybean (Glycine max (L.) Merr) embryos, or isolated cotyledons on MS modified medium supplemented with NAA and 2,4-D, BAP and ABA. When NAA and 2,4-D were compared at similar concentrations (25 and 23 μM), 2,4-D produced larger number of somatic embryos, however, embryogenesis efficiency was improved in media containing NAA by using higher levels (100–150 μM) of the auxin. Somatic embryo morphology varied with auxin type: NAA-induced embryos more closely resembled zygotic embryos than did 2,4-D-induced embryos. Additions of BAP or ABA to auxin-containing media had either no effect or reduced embryo production, although ABA altered the morphology of 2,4-D-induced embryos. In media containing both NAA and 2,4-D, the latter was dominant in terms of embryo morphology. The effects of subculture frequency and of transfers between 2,4-D and NAA media were investigated: Subculture frequency influenced mainly the frequency of normal embryos, while preculture on 2,4-D increased subsequent embryogenesis efficiency on NAA medium but reduced the frequency of normal embryos.
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  • 19
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    Plant cell, tissue and organ culture 10 (1987), S. 209-220 
    ISSN: 1573-5044
    Keywords: soybean ; tissue culture ; somatic embryogenesis ; nutritional and physical factors ; plant regeneration ; plantlet growth
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Immature soybean (Glycine max (L.) Merr) embryos, or cotyledons isolated from them, were cultured on modified MS medium containing B5 vitamins and NAA (50 μM) to induce somatic embryogenesis. The effects of media variables, dissection treatments and light conditions were investigated in this system. The efficiency of embryogenesis increased as sugar concentration decreased from 12 to 1.5%; sucrose and glucose were similarly effective as carbon sources. In an examination of the effects of medium pH, values between pH 5.0 and 7.0 gave similar embryogenesis efficiencies, but the frequency of normal embryos was greater in media with low pH values. In buffered medium (10 mM MES), a pH of 5.0 was inhibitory to embryogenesis, and most normal embryos were produced at pH 5.5. Under various dissection treatments, embryogenesis efficiency and root and callus production were increased by tissue damage. Somatic embryogenesis was observed both in darkness and in light, although embryo development was impaired under high light (80 μE m-2 s-1) conditions. The ability of somatic embryos to germinate was closely correlated with embryo normality, and was influenced little by the hormone content of germination media. Of various media tested for their ability to support the growth of germinated embryos, a medium based on hydroponic nutrient salts, supplemented with yeast extract, and gelled with Difco-Bacto agar gave the best plantlet growth.
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  • 20
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    Plant cell, tissue and organ culture 11 (1987), S. 133-139 
    ISSN: 1573-5044
    Keywords: Rumex acetosella L. ; somatic embryogenesis ; sucrose concentration ; gibberellic acid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Axillary buds of the dioecious plant Rumex acetosella L. were isolated and cultured in vitro. The callus tissue which developed at the basal parts of the explants displayed a high capacity for shoot formation. This morphogenetic pattern was predominant on Murashige and Skoog (MS) medium supplemented with 2% sucrose, 2.2 mgl-1 benzylaminopurine and 0.17 mgl-1 indole-3-acetic acid. Somatic embryogenesis was induced when the osmolality of the medium was increased by adding 6% sucrose instead of 2%, or hexitols in addition to 2% sucrose. Most of the embryogenic calli were formed on the basal parts of leaf laminae and bracts. Development and maturation was strongly promoted by transferring the tissue to a solid or liquid medium lacking benzylaminopurine and indole-3-acetic acid and supplemented with 10 mgl-1 gibberellic acid. The embryos germinated and developed into normal rosette plants when transferred to vermiculite moistened with hormone-free, half-strength MS salt solution. The histology of successive embryogenic stages is presented.
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  • 21
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    Plant cell, tissue and organ culture 11 (1987), S. 189-207 
    ISSN: 1573-5044
    Keywords: somatic embryogenesis ; photomorphogenesis ; ABA ; tissue culture ; Daucus carota ; light quality
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Carrot cells were cultured under various light spectra and intensities at different times following the initiation of suspension cultures from callus. The highest intensity white and blue light treatments were inhibitory to growth and somatic embryogenesis. Red and green light were not different from dark treatments which produced the highest total number of embryoids. After extended time in culture, carrot cells in blue light produced secondary embryoids and anthocyanin. Cultures in red light had multiple cotyledons and orange-pigmented radicles. Leafy cotyledons occurred in all light treatments. Abscisic acid production peaked at the heart stage of embryogenesis and synthesis was most pronounced in blue light. Red light enhanced development to the heart stage. Both the red and blue light spectra may be used to manipulate carrot cell cultures to optimize growth.
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  • 22
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    Plant cell, tissue and organ culture 11 (1987), S. 221-226 
    ISSN: 1573-5044
    Keywords: Triticum aestivum ; callus induction ; plant regeneration ; somatic embryogenesis ; organogenesis ; inflorescence culture ; haploid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Callus cultures were initiated from young inforescences of 7 haploid pollen plants of wheat. The calli from 3 plants were embryogenic and organogenic. After one and a half years of subculture one callus cultures still retained the ability for high frequency of plant regeneration via somatic embryogenesis and sometimes directly differentiated ovary-like structures. All regenerated plants were haploid having 21 chromosomes.
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  • 23
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    Plant cell, tissue and organ culture 12 (1988), S. 31-42 
    ISSN: 1573-5044
    Keywords: Cotton ; Gossypium hirsutum ; somatic embryogenesis ; Coker 312 ; regeneration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Optimal media for induction of somatic embryogenesis from mature and immature tissues ofG. hirsutum L. cv Coker 312 were determined. Explants of three-day-old seedlings form somatic embryos in 100% of cultures when treated with 0.1 mg/1 2,4-dichlorophenoxyacetic acid plus 0.5 mg/1 kinetin. Mature tissues are more recalcitrant than immature tissues and formed somatic embryos on a limited number of hormone treatments. Stem tissue is most readily induced to form somatic embryos by 2 mg/1 napthaleneacetic acid plus 0.1 mg/1 kinetin, whereas leaf tissue formed embryos best when treated with 0.1 mg/1 2,4-dichlorophenoxyacetic acid plus 1.0 mg/1 (2-isopentyl)-adenine, or 1.0 mg/1 napthaleneacetic acid plus 0.5 mg/1 (2-isopentyl)-adenine.
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  • 24
    ISSN: 1573-5044
    Keywords: Bothriochloa ischaemum ; Cynodon dactylon ; forage grasses ; auxin ; tissue culture ; somatic embryogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Objectives of this research were to test the effects of plant genotypes and auxin 2,4-D (2,4-dichlorophenoxyacetic acid) medium concentrations on embryogenic (E) callus production of two grass species. Two Old World bluestem,Bothriochloa ischaemum, accessions (A-8793 and A-8911c) and three bermudagrass,Cynodon dactylon (L.) Pers., accessions (A-10978b, A12164, and ‘Brazos’) supplied the explant material. Immature inflorescences ≤9 mm in length were placed on modified Murashige-Skoog (MS) agar medium containing 0, 1, 3, or 5 mg L-1 of 2,4-D. Explants of all genotypes produced callus by the end of a 4-week dark incubation period at 25°C. When subcultured onto fresh media and maintained at 25°C with a 16 hr photoperiod, calli became embryogenic within 8 weeks of inoculation. Three mg L-1 of 2,4-D in the media maximized E callus production in both bluestem genotypes and in A-10978b and A-12164 bermudagrass genotypes. Maximum E callus production from Brazos bermudagrass resulted from the 1 mg L-1 treatment. Somatic embryos developed after subculture under light. Embryos showed scutellum-like structures and coleoptile-coleorhiza bipolar organization. Plantlets were regenerated from all genotypes except Brazos, whose embryoids failed to germinate. All callus from Brazos eventually senesced. Light and scanning electron microscopy confirmed regeneration through somatic embryogenesis.
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  • 25
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    Plant cell, tissue and organ culture 12 (1988), S. 43-53 
    ISSN: 1573-5044
    Keywords: Cotton ; somatic embryogenesis ; Coker 312 ; regeneration ; Gossypium hirsutum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Calli of cotton (Gossypium hirsutum L.) initiated from seedling hypocotyl tissue were placed in liquid suspension and maintained by serial subculture in hormone-free Murashige and Skoog (MS) medium. Suspensions were sieved and globular embryos collected, washed, resuspended in basal medium and plated onto various semi-solid media. High inorganic salts (MS), low salt (2/3 MS), excess KNO3, and the growth regulators napthaleneacetic acid (NAA), gibberellic acid (GA3) and kinetin were tested for their effects on somatic embryo maturation. Long-term embryo proliferation and maturation were best on medium containing MS plus 1.9g/l KNO3. Embryos 3 mm to 10 mm in size were removed from this plating medium and placed on sterile vermiculite saturated with Stewart and Hsu's medium plus 0.1 mg/l indoleacetic acid (IAA). Plants were recovered from 10.6% of the embryos. When ≥5 mm embryos were placed on this medium, 30% of the embryos formed plants within six weeks. Smaller embryos required a longer period of development on the vermiculite and the addition of fresh medium supplemented with 0.1 mg/l GA3. Plants with an extensive root system and two true leaves were removed from sterile culture and potted in either one-to-one peat and sand, or vermiculite. Eighty percent of the regenerants were successfully hardened when glass beakers of increasing size (10 to 150 ml) were sequentially placed over the young plants during a two-week period.
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    Plant cell, tissue and organ culture 12 (1988), S. 61-66 
    ISSN: 1573-5044
    Keywords: Persea americana ; somatic embryogenesis ; avocado ; picloram
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    Topics: Biology
    Notes: Abstract A tissue culture procedure was developed for the regeneration of somatic embryos from callus cultures of the avocado,Persea americana. Immature zygotic embryos, 0.6–0.8 mm long, were used as original explants. Addition of 0.1 mg/l picloram (4-amino-3,5,6-trichloropicolinic acid) to culture medium appeared critical for callus initiation. Development of somatic embryos was accomplished in picloram concentrations of 0.01 to 0.1 ml/l. A few well developed embryos produced green shoots. Attempts to induce a higher incidence of germination were unsuccessful.
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  • 27
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    Plant cell, tissue and organ culture 12 (1988), S. 83-95 
    ISSN: 1573-5044
    Keywords: Wheat ; callus formation ; somatic embryogenesis ; 3,6-dichloro-o-anisic acid ; 2,4-dichlorophenoxyacetic acid ; 6-furfurylaminopurine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Somatic embryo (embryoid) formation from immature-embryo-derived calli was quantified in replicated experiments involving 10Triticum aestivum L. genotypes. Several published media formulations, which had previously been optimized for wheat tissue culture, were tested for each genotype. Embryos from each plant were randomly assigned to each medium. Percentage precocious germination of immature embryos and mean percentage scutellar callus per explant were recorded. Embryoids per callus were determined by microscopic examination at 28 and 56 days. There were highly significant differences among genotypes, media, and individual plants from which explants were taken. A medium based on double the Murashige and Skoog (MS) inorganic salt concentration was significantly better than other media. Inclusion of all MS vitamins appeared essential for optimal response. Two genotypes were tested in a second experiment where both 3,6-dichloro-o-anisic acid (9.05 μM) and 6-furfurylaminopurine (0.46 μM) were substituted for 2,4-dichlorophenoxyacetic acid (4.52 μM) in either double or normal MS medium. This substitution significantly increased embryoid formation at 28 days. Additions of either 6-furfurylaminopurine or coconut water increased precocious germination of both embryo explants and embryoids.
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  • 28
    ISSN: 1573-5044
    Keywords: wheat ; somatic embryogenesis ; embryogenic callus ; 3,6-dichloro-o-anisic acid ; 2,4-dichlorophenoxyacetic acid ; 6-furfurylaminopurine
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    Notes: Abstract Nine experiments were conducted to determine effects of various culture medium addenda on induction of embryogenic calli from immature embryos of a responsiveTriticum aestivum L. genotype (PCYT 10). Effects were quatified by counting somatic embryos (embryoids) per callus. Optimal auxin concentrations to induce and maintain somatic embryogenesis were 3.62 μM 2,4-dichlorophenoxyacetic acid (2,4-D) or 9.05 μM 3,6-dichloro-o-anisic acid (dicamba). In general, dicamba permitted formation of significantly more embryoids than 2,4-D. Kinetin (6-furfurylaminopurine) at 2.56 μM or 4.65 μM significantly increased percentage scutellar callus when added to 2,4-D or dicamba-containing medium, respectively. Kinetin at 4.65 μM signficantly increased the numbers of embryoids formed when added to medium containing either synthetic auxin. Significantly fewer embryoids formed when cultures were incubated under diffuse light (16-h photoperiod). Casein hydrolysate (200 mgl-1) or L-arginine (0.23 mM) had no effect on numbers of embryoids formed, whereas L-tryptophan (0.20 mM) enhanced such formation with 2,4-D and decreased such formation with dicamba. Two additional experiments generally demonstrated that response to auxin source in the genotypes ND 7532, PCYT 20, Yaqui 50, and Oasis was similar to that in PCYT 10. The higher molar concentration of dicamba required to induce embryogenic callus coupled with more evident embryoid precocious germination and a more rapid rate of tissue necrosis upon extended incubation without subculture suggests that dicamba is metabolized more rapidly than 2,4-D inT. aestivum callus cultures.
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    Plant cell, tissue and organ culture 14 (1988), S. 71-88 
    ISSN: 1573-5044
    Keywords: Gramineae ; sugarcane ; Saccharum ; in vitro ; somatic embryogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Calli have been initiated from young leaves of in vitro grown sugarcane shoots. Histological examination has shown that the two types of calli induced (nodular and friable) originated from different regions of the explants and were cytologically different. This study has shown an obvious relation between the developmental stage of the excised tissue and the potential of plant regeneration of the in vitro initiated callus culture. Nodular calli were obtained from bases of the fast-growing young leaves while their more mature parts of the older leaves only produced friable calli. High-frequency plant regeneration via somatic embryogenesis was obtained from nodular calli while friable calli rarely produced plantlets.
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  • 30
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    Plant cell, tissue and organ culture 14 (1988), S. 207-214 
    ISSN: 1573-5044
    Keywords: olive ; somatic embryogenesis ; zygotic embryos ; leaf discs ; olive tissue culture
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    Topics: Biology
    Notes: Abstract Leaf discs from olive (Olea europaea L.) grown in vitro and immature zygotic embryos collected at 50, 75, 90 and 105 days after full bloom were tested for their somatic embryogenic capacity. The embryos were grown in half-strength MS medium and half-strength OM medium with BAP combinated with either 2,4-D or NAA. Incubation was either in an initial dark period followed by 16h daylight or in 16h daylight throughout. Somatic embryogenesis, approx. 40%, mostly directly from the embryos, was observed only in 75-day-old embryos in medium containing low cytokinin and auxin concentrations. Differentiation was inhibited by 2,4-D whereas NAA did not. In leaf discs and younger and older zygotic embryos, only callus and root formation was observed. Somatic embryos were germinated and then potted-up to soil.
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    Plant cell, tissue and organ culture 15 (1988), S. 33-45 
    ISSN: 1573-5044
    Keywords: somatic embryogenesis ; tissue culture ; histology ; Trifolium ; zygotic embryogenesis ; regeneration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The origin and development of zygotic and somatic embryos of Trifolium rubens L. was studied with the aid of paraffin sections and light microscopy. Zygotic embryos were collected, fixed and prepared daily from one to ten days after cross-pollination. Somatic embryos were obtained by plating petiole sections on modified L2 medium with 0.015 mgl-1 picloram and 0.1 mgl-1 6-BAP. Cultured petioles were collected and fixed daily from one to 25 days after plating. Two regions in the vascular bundle sheath of cultured petioles gave rise to callus. The first region was adjacent to the phloem fibers and produced friable callus. The second region gave rise to compact callus that was connected to the fascicular cambium. Somatic embryos originated from single cells in the cortex directly without intervening callus formation and from single cells in the friable callus. In addition, embryos arose from meristematic regions in compact callus. Many early stages of embryogenesis (one, two and four-celled stages) were observed in the cortex and friable callus. Zygotic embryogenesis in Trifolium differs from other legumes in that the suspensor is short and has a broad attachment. This arrangement was observed in zygotic embryos of T. rubens and in many somatic embryos. However, a continuum of somatic embryogenesis was observed where some young embryos had a Trifolium suspensor-like arrangement while others were attached to a long narrow suspensor-like structure more characteristic of Medicago.
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    Plant cell, tissue and organ culture 15 (1988), S. 67-71 
    ISSN: 1573-5044
    Keywords: minimum-growth storage ; cold storage ; silicone ; low temperature ; Vitis ; tissue culture ; regeneration ; somatic embryogenesis
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    Topics: Biology
    Notes: Abstract Effects of the combination of low temperature and silicone treatment on the storage of grape callus (Vitis vinifera L. x V. labrusca L. cv. Kyoho; V. vinifera L. cv. Koshusanjaku) were examined. In ‘Kyoho’, the calli were stored at 10°C successfully for up to 360 days. Embryogenic calli of ‘Koshusanjaku’ stored at 10°C retained the ability of embryogenesis after 360 days of storage. However, the color of both calli became brownish. This was improved by the combination of low temperature and silicone treatment. The calli of ‘Kyoho’ survived by the storage under the combination of 15°C and silicone. Embryogenic calli stored at 10 and 15°C in combination with silicone survived for 360 days, and regenerated only after transfer onto a regeneration medium. Thus the combination of low temperature and silicone affects the longevity of the grape callus.
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  • 33
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    Plant cell, tissue and organ culture 15 (1988), S. 161-167 
    ISSN: 1573-5044
    Keywords: Cyphomandra betacea ; somatic embryogenesis ; plant regeneration ; tissue culture ; growth regulators
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    Notes: Abstract Callus cultures with globular proembryogenic structures were induced from zygotic embryos and hypocotyl segments of Cyphomandra betacea on MS medium supplemented with 2,4-D. Proembryogenic structures produced somatic embryos and plantlets on regulator-free basal medium. Pieces of embryogenic callus subcultured on medium with the same original composition gave rise to new globular structures and the potential for plantlet regeneration has been maintained for over a year. The histological examination of these proembryogenic structures suggested that somatic embryos arise from single cells. Regenerated plants are phenotypically normal, having diploid chromosome numbers (2n = 24).
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  • 34
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    Plant cell, tissue and organ culture 16 (1989), S. 103-111 
    ISSN: 1573-5044
    Keywords: somatic embryogenesis ; regeneration ; tissue culture ; Picea abies ; glutamine ; organic nitrogen
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    Topics: Biology
    Notes: Abstract Somatic embryos and rooted plantlets have been regenerated from light-initiated embryogenic callus derived from mature embryos of Picea abies. Under a 16 h photoperiod, mature zygotic embryos were cultured on a modified half-strength Murashige & Skoog medium without NH4NO3 and supplemented with 5 mM glutamine, 4.5 μM N6-benzyladenine and 10.7 μM naphthaleneacetic acid or 10 μM 2,4-dichlorophenoxyacetic acid. White translucent embryogenic callus, proliferating from the callusing hypocotyl region after 3 weeks incubation, was isolated from the green non-embryogenic tissue and subcultured for over 12 months. Upon transfer of the embryogenic callus through a specific sequence of media, somatic embryos proceeded to mature, elongating and forming rings of cotyledonary leaves similar to those of zygotic embryos. Transferred to medium without growth regulators, the somatic embryos ‘germinated’ and produced plantlets with green cotyledons, elongated hypocotyls and primary roots.
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  • 35
    ISSN: 1573-5044
    Keywords: Dioscorea composita ; Dioscorea cayenensis ; yam ; somatic embryogenesis ; plant regeneration ; mature zygotic embryos ; micropropagation
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    Topics: Biology
    Notes: Abstract A tissue culture method for regeneration of plantlets from calluses of Dioscorea composita Hemsl. and Dioscorea cayenensis L. is described. Zygotic embryos were used as initial explants. Calluses were obtained on Murashige & Skoog basal medium supplemented with 18 μM 2,4-D and plantlets were regenerated on media containing 0.1 μM zeatin and 3.3 mM glutamine according to previously described protocols [3]. Inclusion of 0.3% (w/v) activated charcoal in media did not increase callusing. Regeneration of plantlets from D. cayenensis calluses occurred only at low levels of 2,4-D (2.25 μM) contained in the media tested. The results indicated that there were genotype-dependent differences between the yam species in their ability to regenerate plantlets in vitro.
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  • 36
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    Plant cell, tissue and organ culture 18 (1989), S. 143-151 
    ISSN: 1573-5044
    Keywords: Zea mays L. ; tissue culture ; somatic embryogenesis ; type II callus ; silver nitrate, ethylene
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    Topics: Biology
    Notes: Abstract In order to evaluate the impact of ethylene in maize tissue culture, silver nitrate has been used as an inhibitor of ethylene action. Type II callus initiation rate was improved when immature embryos were cultured on a modified Murashige & Skoog medium containing various concentrations of silver nitrate (5, 10, 20 mgl-1). Regeneration ability of calli initiated and maintained in presence of silver nitrate was enhanced. No modification of callus growth rate neither of ethylene production has been detected.
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    Plant cell, tissue and organ culture 18 (1989), S. 181-189 
    ISSN: 1573-5044
    Keywords: celery ; mannitol ; somatic embryogenesis ; singular somatic embryos ; cell culture
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    Topics: Biology
    Notes: Abstract Embryogenic suspension cultures of celery (Apium graveolens L.) were established from petiole and leaf callus. Suspensions were routinely subcultured in a ‘maintenance medium’ (with 2.3 μM 2,4-D and 0.88 μM BA). Somatic embryogenesis occurred in this medium, but was considerably improved in a ‘regeneration medium’ (2.3 μM kinetin, without 2,4-D). Cultures thus maintained, contained embryogenic clumps, aggregated somatic embryos, and few free-floating singular somatic embryos. Addition of mannitol (3–4% w/v) prevented cell lysis, greatly increased the number of singular somatic embryos, improved their normal differentiation, and accelerated torpedo embryo development. Experiments to reveal the nature of the mannitol effect demonstrated that the decreased osmotic potential was an important factor, but not the only one: iso-molar solutions of sucrose alone were not as effective. The mannitol effect could be manifested after a short (2–3 days) exposure period, suggesting a ‘trigger’ (induction) mechanism. Several pathways of somatic embryogenesis in celery and its regulation by subculturing, with the addition of mannitol, are outlined. Cultures thus maintained resulted in a high rate of normal somatic embryogenesis and production of normal transplantable celery plants.
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    Plant cell, tissue and organ culture 19 (1989), S. 113-127 
    ISSN: 1573-5044
    Keywords: somatic embryogenesis ; plant regeneration ; protoplasts ; Trifolium pratense ; red clover ; protoclonal variation
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    Topics: Biology
    Notes: Abstract Protoplasts are useful for subcellular studies, in vitro selection, somatic hybridization and transformation. Whole plant regeneration from protoplasts is a prerequisite to producing altered crop plants using these methods. Whole plant regeneration was achieved from leaf- and suspension culture-derived protoplasts of T. pratense. Regeneration was most dependent upon identifying genotypes with genetic capacity to regenerate. Additional factors that were used to select genotypes, but which proved to be less important, were a high rate of cell growth in culture and a high plating efficiency of protoplasts. One genotype was identified which had a regeneration response equivalent to that of T. rubens and which regenerated from both leaf- and suspension culture-derived protoplasts.
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    Plant cell, tissue and organ culture 19 (1989), S. 167-174 
    ISSN: 1573-5044
    Keywords: P. coccineus ; in vitro culture ; immature cotyledons ; organogenesis ; somatic embryogenesis
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    Topics: Biology
    Notes: Abstract A method for plant regeneration, via organogenesis and somatic embryogenesis, from P. coccineus is described. Immature cotyledons from plants regenerated and cloned in previous experiments were used. The highest percentage of regeneration (37.5%) was observed from the cotyledons of clone C7 on a modified Murashige & Skoog basal medium to which (2-isopentenyl)adenine (2iP, 10 mg l-1) and 2-naphthoxyacetic acid (NOA, 0.05 mg l-1) were added. On average, the same medium gave the highest percentage of regeneration also from the cotyledons of the 7 clones of P. coccineus used in the experiment. Newly regenerated plants were normally fertile. The procedure described may serve for induction of somaclonal variation for in vitro selection of valuable genotypes and for genetic transformation by A. tumefaciens.
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  • 40
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    Methods in cell science 12 (1989), S. 179-183 
    ISSN: 1573-0603
    Keywords: artificial seeds ; somatic embryogenesis ; alfalfa ; Medicago sativa L.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A detailed description of alfalfa (Medicago sativa L.) somatic embryogenesis and methods to produce alfalfa artificial seeds are described. These protocols provide directions for producing alfalfa artificial seeds that will convert into whole plants when planted in vitro or directly into soil.
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    Plant cell, tissue and organ culture 4 (1985), S. 111-122 
    ISSN: 1573-5044
    Keywords: alfalfa ; somatic embryogenesis ; germplasm ; genotypic variation
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    Topics: Biology
    Notes: Abstract Seventy-six cultivars of alfalfa (Medicago sativa L., M. falcata L. and M. varia Martyn) were tested in vitro for their capacity to produce callus and somatic embryos. A three-step media protocol was used to survey the response of the cotyledons and hypocotyl of each genotype while the epicotyl region was conserved in order to recover highly responding genotypes. The best regeneration response was observed in creepingrooted cultivars which contained a strong genetic contribution of two landrace germplasm sources, defined as M. falcata and Ladak, in their ancestry. The callus and embryogenesis responses showed a high degree of variation both between cultivars and among the plants of many of the 76 cultivars tested. A higher number of plants produced somatic embryos in the high regenerating cultivars compared to the low regenerating cultivars regardless of the media protocol or explant.
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  • 42
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    Plant cell, tissue and organ culture 4 (1985), S. 123-133 
    ISSN: 1573-5044
    Keywords: Dactylis glomerata ; orchardgrass ; suspension culture ; somatic embryogenesis ; dicamba ; casein hydrolysate
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    Topics: Biology
    Notes: Abstract The effects of various concentrations and combinations of dicamba (3,6-dichloro-o-anisic acid) and casein hydrolysate on growth, mucilage accumulation, somatic embryo and root development in suspension cultures of Dactylis glomerata L. (orchardgrass) were examined. Fresh weight of culture tissue was increased with 20 μM but not with 80 or 160 μM dicamba in treatments with 1–4 g/l casein hydrolysate. Different casein hydrolysate concentrations did not alter the amount of mucilage (measured by viscosity) in the supernatant in the absence of dicamba. However, the addition of dicamba increased viscosity with 80 μM giving the maximum response. Casein hydrolysate produced the greatest viscosity at 1–3 g/l in treatments where dicamba was present. Both dicamba and casein hydrolysate were required for development of somatic embryos. Dicamba at 40 μM with 3–4 g/l casein hydrolysate produced approximately 2000 embryos/35 ml of suspension. Root development was inhibited by dicamba and stimulated by the presence of casein hydrolysate. The usefulness of medium component manipulations for influencing somatic embryogenesis and culture quality is discussed.
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  • 43
    ISSN: 1573-5060
    Keywords: Triticum durum ; durum wheat ; callus ; organogenesis ; plantlet regeneration ; somatic embryogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Experiments upon in vitro culture of immature durum wheat embryos, harvested at different growth stages, were made in two consecutive years. Callus formation and plantlet regeneration were obtained. The ability to form callus and the degree of morphogenetic processes varied with the different hormonal treatments used and with the age of the embryos. In the first year the best response for callus growth was observed with 2,4-D 2 mg l-1 plus adenine 50 mg l-1 or 2,4-D 5 mg l-1 alone in the more mature embryos (15 and 20 days after anthesis). On the contrary, NAA 5 mg l-1 had a greater shoot regeneration effect. In the next year, at all 2,4-D concentrations and for the two different ages of the embryos tested, all embryos formed callus. Regeneration of plantlets was obtained in higher percentage in calli originated from the more developed embryos. The effect of changed media upon plantlet regeneration was studied after callus transplant. Investigation by cytophotometry and chromosome counts on different calli showed, practically in all cells, a diploid condition. A histological analysis demonstrated embryogenic somatic characteristics in many samples of callus. The pattern of organogenesis seemed to be via adventitious bud formation but structures resembling embryoids were also observed in the callus.
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    Plant cell, tissue and organ culture 6 (1986), S. 139-147 
    ISSN: 1573-5044
    Keywords: Citrus sinensis ; somatic embryogenesis ; growth regulator
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    Topics: Biology
    Notes: Abstract Embryogenic cultures were initiated from undeveloped ovules of several polyembryonic Citrus species on a basal medium supplemented with either malt extract, 2,4-D alone, or 2,4-D in combination with BA or daminozide. Primary embryos of all responsive cultivars were harvested directly from ovule cultures; secondary embryo harvests were made from ‘Handin’ orange ovule cultures and long-term embryogenic callus. Differences were observed among cultivars and treatments in percentage of responsive ovules and total number of embryos produced. The most effective treatment for embryo production varied among cultivars. Embryo germination and plant establishment frequencies were determined for this plant regeneration system. Differences among cultivars with respect to regenerate survival percentage were minimal. Plant regeneration via secondary or long-term callus-derived embryos was as efficient as from primary embryos. Critical factors influencing plant production and survival were the production of normal viable embryos, balanced germination, and successful acclimatization to the external environment.
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    Plant cell, tissue and organ culture 8 (1987), S. 73-81 
    ISSN: 1573-5044
    Keywords: alfalfa ; Medicago ; somatic embryogenesis ; genotypic variation
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    Topics: Biology
    Notes: Abstract Fifty genotypes of each of three cultivars of alfalfa (Medicago spp.) were tested in three medium protocols for their capacity to produce somatic embryos and plantlets from callus cultures. Highly productive genotypes produced somatic embryos regardless of medium protocol or explant source, while other genotypes produced somatic embryos in a medium-specific or explant-specific fashion. The results showed that embryogenesis in mature leaf-derived calli could be predicted from the frequency of embryo formation in cotyledon-derived calli of the same genotype. The results also indicated that highly productive genotypes can be selected from cultivars with a low frequency of regeneration.
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  • 46
    ISSN: 1573-5044
    Keywords: Coffea canephora ; coffee ; protoplast ; somatic embryogenesis ; cell suspension
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    Notes: Abstract Somatic embryogenesis was achieved in protoplast cultures of coffee. Protoplasts were isolated from cell suspension-derived somatic embryos of Coffea canephora. After repeated subculture in a medium supplemented with 0.5 mg/l of each of kinetin, 2,4-dichlorophenoxyacetic acid (2,4-D), and naphthaleneacetic acid (NAA), microcalli developed. Transfer of these microcalli to a medium lacking growth regulators resulted in the formation of globular embryos. Upon subculture without growth regulators they grew to well-differentiated embryos, Eventually some of them developed to plantlets which were transferred to the greenhouse for further observation.
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    Plant cell, tissue and organ culture 9 (1987), S. 73-80 
    ISSN: 1573-5044
    Keywords: grape ; secondary embryogenesis ; somatic embryogenesis ; somatic embryo ontogeny ; tissue culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Anthers and ovaries of Vitis longii ‘Microsperma’ produced embryogenic callus when cultured on solidified Murashige and Skoog medium with 5μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 1μM benzyladenine (BA). The initial callus was short-lived. However, long-term embryogenesis from callus was maintained through serial transfers by careful selection of clustered embryos with subtending callus. Alternatively, long term culture maintenance was through secondary embryogenesis which occurred directly from previously formed embryos on medium lacking growth regulators. Somatic embryos were white, exhibited frequent pluricotyly and tended to be larger than zygotic embryos. Histology of embryogenic callus demonstrated the presence of lipid-like substances and abundant starch. Somatic embryos were attached to callus by narrow to wide suspensor-like structures and possessed typical epidermal, cortical, and vascular tissue. Embryo cells contained abundant lipid-like accumulations but no starch. Embryos germinated when placed on medium containing 1μM BA and produced plants of normal appearance.
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  • 48
    ISSN: 1573-5060
    Keywords: Cucumis metuliferus ; Cucumis anguria ; embryo culture ; somatic embryogenesis ; interspecific hybridization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Interspecific crosses between Cucumis metuliferus Naud. and C. anguria L. were obtained through embryo culture. Embryos in the rabbit-ear to advanced fluke-shaped stages were rescued 34–99 days after pollination. Plants were obtained through direct embryo culture, and through somatic embryogenesis from immature embryos. For direct embryo culture, fluke-shaped embryos were stored in sterile water in darkness for three days at 25C prior to transfer on Murashige and Skoog (MS) culture medium plus 1.0 μM 6-benzylamino-purine. Multiple plants were obtained from single embryos through somatic embryogenesis of rabbit-ear stages on MS plus 10 μM indole-3-acetic acid and 5 μM 6-benzylamino-purine. Evidence of hybridization included leaf shape intermediate between the two parents, penduncle shape prior to fertilization which resembled the male parent, low pollen viability and isoelectric focussing of protein bands for acid phosphatase of leaf extracts.
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  • 49
    ISSN: 1573-5060
    Keywords: callus culture ; organogenesis ; pea ; Pisum sativum ; somaclonal variation ; somatic embryogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The possibility of producing agronomically-useful somaclones via organogenesis and somatic embryogenesis from callus cultures of pea (Pisum sativum L.) was studied. Organogenic calli were induced from immature leaflets on MSB medium with NAA and BAP. Embryogenic calli were derived either from immature zygotic embryos (using 2,4-D) or from shoot apices (using picloram) of aseptically-germinated seedlings. The seed progenies (T1 to T3-generation) of primary regenerants were grown in field conditions and their phenotypic variation was evaluated and compared with control, non-tissue culture-derived plant material. In addition, electrophoretic analyses of selected isoenzyme systems and total proteins have been done. The results do not show dramatic changes in qualitative and quantitative traits. The evaluation of at least two future generations (T4, T5) is planned.
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  • 50
    ISSN: 1573-5060
    Keywords: Picea abies ; genetic stability ; somatic embryogenesis ; RAPD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Somaclonal variation, which is a welcome source of genetic variation for crop breeding, is unwanted when direct regenerants have to be used in tissue culture mass propagation (eg. in many forest trees), or in the regeneration of genetically transformed plants. Random amplified polymorphic DNA (RAPD) was used to analyse somatic embryos and plants regenerated from embryogenic cell lines in Norway spruce, Picea abies (L.) Karst. RAPD facilitated the identification of clones, as material from the same cell lines shared identical patterns of amplified fragments, whereas regenerants from different cell lines were easily distinguishable by their respective patterns. For comparisons with explant donor genotypes, cell lines were initiated from cotyledons. Some of the seedlings that had parts of their cotyledons removed were grown on as control plants. Somatic embryos regenerated from cotyledon cell lines showed no aberrations in RAPD banding patterns with respect to donor plants. We conclude that gross somaclonal variation is absent in our plant regeneration system.
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  • 51
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    Plant cell, tissue and organ culture 19 (1989), S. 243-256 
    ISSN: 1573-5044
    Keywords: callus histology ; Hevea brasiliensis ; rubber tree ; somatic embryogenesis ; subculture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Somatic embryos ofHevea brasiliensis can be obtained by culturing thin sections of inner tegument of seed on two successive different media, MH1 and MH3. Histological study showed that in calli cultured on non-renewed medium MH1 for 40 days, the embryogenesis process initiated on the 20th day did not produce results owing to early degeneration of the cells involved in the embryogenic pathway. However, typical embryogenic cells formed when medium MH1 was renewed once during the first phase of culture (between day 20 and day 30). Proembryos developed when the calli were subcultured on medium MH3 10–15 days later. Embryogenic cells did not form when there was frequent renewal of medium MH1 or early subculturing on MH3 after less than 40 days of culture on MH1. Methodical histological monitoring of the development of embryogenic quality of calli thus made it possible to define the optimum culture sequences for the embryogenesis process and which are favourable for regular obtaining of proembryos.
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  • 52
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    Plant cell, tissue and organ culture 17 (1989), S. 53-58 
    ISSN: 1573-5044
    Keywords: Abies alba ; European silver fir ; somatic embryogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Megagametophytes of Abies alba containing the immature embryos were dissected from the seed coats and divided by longitudinal and transverse sections. They were placed with the cut surface down on modified Schenk & Hildebrandt medium containing 50 mgl-1 myo-inositol and 2% sucrose, supplemented with 1 mgl-1 N6-benzyladenine (BAP). An embryogenic type of callus proliferated after one month of culture. Closer examination revealed the presence of structures resembling early stages of embryogenesis as well as of single elongated, vacuolated cells and clusters of cells with dense cytoplasm. Under appropriate conditions, some of the somatic embryos elongated and formed cotyledons.
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  • 53
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    Plant cell, tissue and organ culture 17 (1989), S. 39-52 
    ISSN: 1573-5044
    Keywords: sweet potato ; somatic embryogenesis ; 2,4-dichlorophenoxyacetic acid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Embryogenic suspension cultures of Ipomoea batatas Poir. contain heterogeneous populations of discrete cellular units. In order to optimize embryo production, a study was conducted to identify the embryogenic fraction of such cultures. Suspension cultures were fractionated with sieves of 1000, 710, 500, 355, 250, 180, 125, 90 and 63μm mesh openings and the composition of each fraction was determined. Cellular units larger than 355 μm were primarily calli and made up 75% of the total mass of cultures in the stationary phase of growth. These calli were composed of embryogenic and non-embryogenic subunits, and 98% of the embryogenic subunits measured 355–1000 μm. Calli and embryogenic calli subunits produced clusters of embryos at various stages of development upon transfer to liquid or solidified media without 2,4-D. The 125–355 μm fraction of suspension cultures was composed of cell aggregates of which 20% were embryogenic. The embryogenic cell aggregates produced single globular embryos upon transfer to liquid media containing 0 or 1 μM 2,4-D. The 63–125 μm fraction of suspension cultures contained only 2% of embryogenic cell aggregates. It can be inferred from our results that the embryogenic fraction of cultures was essentially represented in calli, and that proliferation of the embryogenic fraction occurred through the separation of embryogenic cell aggregates from larger calli when cultures approached their stationary growth phase.
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  • 54
    ISSN: 1573-5044
    Keywords: Daucus carota ; esterases ; medium proteins ; PAGE ; somatic embryogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A unique esterase isozyme ‘z’ with very low electrophoretic mobility on the anionic polyacrylamide gel (PAGE) was found in the medium of a non-embryogenic (Ca-4) line of cultured carrot (Daucus carota L.) cells. The protein corresponding to this esterase isozyme ‘z’ was purified by electroelution from preparative PAGE and the esterase migrated as a single band with an apparent M r of 35 000 on SDS-PAGE. The purified esterase isozyme ‘z’ exhibited at least 350-fold higher specific activity than that in the total medium proteins.
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  • 55
    ISSN: 1573-5044
    Keywords: wheat ; Triticum aestivum ; ABA ; ABA analogs ; somatic embryogenesis ; precocious germination
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Zygotic embryos from ten spring wheat (Triticum aestivum L.) genotypes were tested for embryogenic callus induction in the presence or absence of externally supplied (±)-abscisic acid (ABA) and two of its analogs, methyl abscisate and methyl epoxy-beta-ionylideneacetate. (±)-ABA and its analogs suppressed precocious germination of cultured late-stage embryos and promoted embryogenic callus induction. A significantly greater number of plants was regenerated from calli induced in the presence of ABA and ABA analogs. Early-stage embryos when cultured in the presence of (±)-ABA showed a negative response. Possible roles of ABA with respect to the expression of somatic embryogenesis are discussed.
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  • 56
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    Plant cell, tissue and organ culture 3 (1985), S. 51-54 
    ISSN: 1573-5044
    Keywords: Oryza sativa ; rice ; callus ; inflorescence ; somatic embryogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Compact calli initiated from young inflorescences of Oryza sativa L. (rice) on the Linsmaier and Skoog's (LS) medium containing 1 to 2.5mg/l of 2,4-dichlorophen-oxyacetic acid (2,4-D) were used for regeneration studies. After smooth and compact nodules appeared, these calli were transferred to the regeneration medium containing indole-3-acetic acid (IAA) and either kinetin or 6-benzylaminopurine (BAP). Somatic embryos developed in ten days and were examined by histological studies. Some of the embryos showed scutellum-like structures and a coleoptile-coleorhiza bipolar organization. Regenerated plants had the normal chromosome number of 2n=24.
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