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  • 1
    Electronic Resource
    Electronic Resource
    Screening of diploid Medicago sativa germplasm for somatic embryogenesis (1985)
    Springer
    Plant cell reports 4 (1985), S. 285-288 
    Details
    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.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00269379
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  • 2
    Electronic Resource
    Electronic Resource
    Donor tissue and culture condition effects on mesophyll protoplasts of Medicago sativa (1987)
    Springer
    Plant cell, tissue and organ culture 9 (1987), S. 217-228 
    Details
    ISSN: 1573-5044
    Keywords: alfalfa ; pretreatment ; culture medium ; direct embryogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Mesophyll protoplasts were produced from clones of two cultivars of Medicago sativa, ‘Rangelander’ and ‘Regen S’. Protoplasts from the ‘Regen S’ clone generally gave rise to calli while those from the ‘Rangelander’ clone would undergo direct embryogenesis. Effects of plant growth conditions, donor tissue pretreatment and protoplast culture conditions on mesophyll protoplast production and subsequent development patterns were investigated. The major factor determining whether or not mesophyll protoplasts would be produced from either of the clones was the pretreatment in water of shoots excised from the donor plants. Pretreatment in water containing growth regulators did not alter protoplast production or development in the ‘Regen S’ clone. Pretreatment of the ‘Rangelander’ clone shoots with abscisic acid or naphthaleneacetic acid was slightly beneficial to embryo production while pretreatment with benzylaminopurine was detrimental. Altered leaf morphology induced by growth condition changes did not affect mesophyll protoplast production or subsequent development patterns when shoots were pretreated in water. Culture of protoplasts in liquid droplets or solid agar medium increased low density protoplast survival and subsequent embryo production in the ‘Rangelander’ clone.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00040807
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    Paper (German National Licenses)
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  • 3
    Electronic Resource
    Electronic Resource
    Role of exogenous reduced nitrogen and sucrose in rapid high frequency somatic embryogenesis in Medicago sativa (1987)
    Springer
    Plant cell, tissue and organ culture 10 (1987), S. 11-19 
    Details
    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.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00037492
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  • 4
    Electronic Resource
    Electronic Resource
    Role of genetic background in somatic embryogenesis in Medicago (1985)
    Springer
    Plant cell, tissue and organ culture 4 (1985), S. 111-122 
    Details
    ISSN: 1573-5044
    Keywords: alfalfa ; somatic embryogenesis ; germplasm ; genotypic variation
    Source: Springer Online Journal Archives 1860-2000
    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.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00042269
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  • 5
    Electronic Resource
    Electronic Resource
    Development of a simple particle bombardment device for gene transfer into plant cells (1994)
    Springer
    Plant cell, tissue and organ culture 37 (1994), S. 47-53 
    Details
    ISSN: 1573-5044
    Keywords: transient gene expression ; β-glucuronidase ; alfalfa ; soybean ; DNA transfer
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A simple particle bombardment device was designed, constructed and shown to be efficient for the delivery of DNA into plant cells. High levels of transient β-glucuronidase expression were observed in alfalfa suspension-cultured cells and embryogenic soybean suspension-cultured cells. Expression of β-glucuronidase in alfalfa suspension-cultured cells was used to optimize the bombardment conditions for the device. Transient gene expression in alfalfa was found to be dependent on the state of the target tissue, the size of particles employed, the helium pressure used to accelerate the particles and the distance travel led by the tungsten particles carrying DNA.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00048116
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    Paper (German National Licenses)
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  • 6
    Electronic Resource
    Electronic Resource
    Patterns of direct and indirect embryogenesis from mesophyll protoplasts of Medicago sativa (1987)
    Springer
    Plant cell, tissue and organ culture 9 (1987), S. 121-130 
    Details
    ISSN: 1573-5044
    Keywords: Medicago sativa ; direct embryogenesis ; genotypic variation ; mesophyll protoplast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Clones of three cultivars of Medicago sativa (‘Rambler’, ‘Regen S’ and ‘Rangelander’) were used as sources of mesophyll protoplasts. Although all three clones readily produced protoplasts, the subsequent development patterns in culture varied greatly among genotypes, with protoplasts from ‘Regen S’ and ‘Rambler’ forming calli which could be induced to form embryos, and protoplasts from ‘Rangelander’ undergoing direct embryogenesis. Protoplasts of ‘Regen S’ exhibited high rates of division while those of ‘Rangelander’ tended to aggregate with only a few cells per aggregate surviving. The surviving cells gave rise to proembryos within the aggregates; these proembryos developed into differentiated embryos after 5–7 weeks of culture. Based on the initial protoplast population, the efficiency of embryo formation averaged 0.13% and ranged from 0.001–0.4%. Observations during the early stages of culture indicated that cell aggregation was a prerequisite for direct embryogenesis.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00044247
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  • 7
    Electronic Resource
    Electronic Resource
    Plant regeneration from suspension culture and mesophyll protoplasts of Medicago sativa L. (1984)
    Springer
    Plant cell, tissue and organ culture 3 (1984), S. 149-162 
    Details
    ISSN: 1573-5044
    Keywords: alfalfa ; somatic embryogenesis ; protoplast ; cell suspensions
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A system was established for achieving plant regeneration from mesophyll protoplasts and cotyledon-derived cell suspension cultures of alfalfa, Medicago sativa L. Peeled leaflets or cells from 6-day-old cell suspensions were incubated in an enzyme mixture containing 1% Driselase, 1% Rhozyme, 0.1% Cellulase and 72 gl-1 mannitol at pH 5.8 for 2–16 h to liberate protoplasts. A complex Kao medium supported cell division and colony formation, whereas a high auxin/low cytokinin treatment on Schenk and Hildebrandt medium followed by culture on growth regulator-free Blaydes or Linsmaier and Skoog medium resulted in somatic embryo formation. Of the three varieties tested. Citation, Answer and Regen S, the latter two produced embryos from which plants could be regenerated.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00033736
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  • 8
    Electronic Resource
    Electronic Resource
    Characterization of competence during induction of somatic embryogenesis in alfalfa tissue culture (1993)
    Springer
    Plant cell, tissue and organ culture 34 (1993), S. 125-132 
    Details
    ISSN: 1573-5044
    Keywords: 2,4-dichlorophenoxyacetic acid ; Medicago sativa ; phenylacetic acid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Systematic manipulations of the culture protocol leading to somatic embryogenesis in alfalfa petiole-derived callus were performed to study the competence phase during somatic embryogenesis in alfalfa. These demonstrated a requirement for the acquisition of competence prior to the induction of the embryogenesis pathway. Induction was triggered by a number of synthetic auxins including 2,4-dichlorophenoxyacetic acid (2,4-d). Competence could be acquired in the presence of these auxins as well as phenylacetic acid (PAA), an auxin that did not induce embryogenesis. Different degrees of competence were apparently acquired by exposure to 2,4-d and PAA. Some degree of competence was acquired in the absence of auxin treatment. The current understanding of the concept of competence is discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00036092
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