Institut für Landwirtschaftliche Botanik der Universität Bonn, Meckenheimer Allee 176, D-53115, Bonn, Germany.
Plant Cell Rep. 1996 Jun;15(10):742-5. doi: 10.1007/BF00232219.
Hypocotyl protoplasts of four different Helianthus annuus genotypes were cultivated for 22-28 days in agarose droplets covered with liquid medium. In the first week, supplementation of the medium with plant growth regulators was at a 0.8/1 ratio of cytokinin and auxin followed by a high auxin concentration in the second week and a cytokinin to auxin ratio of 8/1 in the third and fourth week. Following transfer onto solid medium containing cytokinin and auxin in a proportion of 40/1 morphogenic callus started to form globular structures that developed into leaf primordia. Subsequent shoot elongation and rooting were obtained on hormone free medium after dipping the cut shoots into high auxin solution. Thirteen weeks after protoplast isolation, plantlets could be transferred to the greenhouse. Shoot regeneration was obtained for all four cultivars (Florom-328, Cerflor, Euroflor, Frankasol) at different rates reflecting their regenerative potential.
将四个不同的向日葵基因型的下胚轴原生质体在琼脂糖液滴中培养 22-28 天，液滴覆盖在液体培养基上。在第一周，培养基中植物生长调节剂的添加比例为细胞分裂素和生长素的 0.8/1，然后在第二周生长素浓度较高，第三和第四周细胞分裂素和生长素的比例为 8/1。继转移到固体培养基上，含有细胞分裂素和生长素的比例为 40/1，形态发生愈伤组织开始形成球形结构，发育成叶原基。将切割的嫩枝浸泡在高浓度生长素溶液中后，在无激素的培养基上可以获得随后的嫩枝伸长和生根。原生质体分离后 13 周，可以将幼苗转移到温室中。所有四个品种（Florom-328、Cerflor、Euroflor、Frankasol）都以不同的再生率获得了芽再生，反映了它们的再生潜力。
