Wardrop J, Davey M R, Power J B, Lowe K C
Department of Life Science, University of Nottingham, U.K.
Artif Cells Blood Substit Immobil Biotechnol. 1997 Sep;25(5):481-6. doi: 10.3109/10731199709118938.
Electrofused Passiflora protoplasts (P. edulis, P. giberti) were plated in KPR medium overlaying oxygen-gassed perfluorodecalin (Flutec PP6). Oxygenated PFC significantly (P < 0.05) enhanced protoplast division, as reflected by an increase in mean plating efficiency of up to 62% (P < 0.05) over 14 days. After 21 days of culture, the liquid phase containing dividing protoplast-derived cells, was removed from the PFC surface and overlaid onto MS-based agar medium for callus proliferation. Forty days later, protoplast-derived calli were transferred to one of two regeneration protocols, previously determined using unfused parental protoplasts. Calli derived from electrofusion-treated protoplasts exhibited organogenesis or somatic embryogenesis, depending on the regeneration procedure. The regeneration efficiency after 121 days for protoplasts initially cultured with oxygenated PFC was over 2-fold greater (P < 0.01) than control. These results indicate that oxygenated PFC can enhance growth and regeneration of protoplasts and their fusion products.
将电融合的西番莲原生质体（西印度醋栗、吉氏西番莲）接种于覆盖充氧全氟萘烷（氟替卡PP6）的KPR培养基中。充氧的全氟化合物显著（P < 0.05）提高了原生质体的分裂能力，这体现在14天内平均接种效率提高了62%（P < 0.05）。培养21天后，将含有正在分裂的原生质体来源细胞的液相从全氟化合物表面移除，并覆盖在基于MS的琼脂培养基上以促进愈伤组织增殖。40天后，将原生质体来源的愈伤组织转移至之前使用未融合的亲本原生质体确定的两种再生方案之一。根据再生程序，电融合处理的原生质体产生的愈伤组织表现出器官发生或体细胞胚胎发生。最初用充氧全氟化合物培养的原生质体在121天后的再生效率比对照高出2倍多（P < 0.01）。这些结果表明，充氧的全氟化合物可以促进原生质体及其融合产物的生长和再生。
