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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Plant cell, tissue and organ culture 9 (1987), S. 217-228 
    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
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Plant cell, tissue and organ culture 9 (1987), S. 121-130 
    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
    Library Location Call Number Volume/Issue/Year Availability
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