Effect of enzyme concentrations on protoplast isolation and protoplast culture of Spathiphyllum and Anthurium
Effect of enzyme concentrations on protoplast isolation and protoplast culture of Spathiphyllum and Anthurium
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DOI：
10.1007/s11240-007-9226-3
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发表时间：
2007-11-01
影响因子：
3
通讯作者：
Van Huylenbroeck, Johan
中科院分区：
文献类型：
--
作者：
Duquenne, Barbara;Eeckhaut, Tom;Van Huylenbroeck, Johan
Vital protoplasts from Spathiphyllum wallisii 'Alain' and Anthurium scherzerianum '238' were isolated from both somatic embryos and leaves. The highest yields were obtained when 1.5% cellulase, 0.5% macerase and 0.5% driselase were used for Spathiphyllum wallisii leaves and 0.5% cellulase, 0.3% macerase and 0.5% driselase for Anthurium scherzerianum embryos. About 1 x10(6) protoplasts g(-1) and 1 x10(5) protoplasts g(-1) could be isolated from leaves and embryos, respectively. For protoplast fusion Spathiphyllum wallisii 'Alain' and Anthurium scherzerianum '238' were mixed in a 1:1 ratio in a fusion solution containing 1 mM CaCl2 center dot 2H(2)O, 1 mM MES and 0.5 M mannitol. Fusion was performed by protoplast alignment under 500 V cm(-1) alternating current for 60 s and subsequent generation of two pulses of 4500 V cm(-1) direct current during 50 mu s. Development until colony stage was achieved using agarose beads for protoplast culture.
