LONG-TERM CONSERVATION AND EFFECTIVE REGENERATION OF PLANT CELLS IN ARTIFICIAL SEED CAPSULES

1.在25ﾟ℃培养条件下，研究了琼脂凝胶含水率和气相氧气浓度对辣根毛状根细胞团(CA)不定根再生的影响。琼脂凝胶含水率为66%时，在培养20天内，CA的生根数高度依赖于琼脂凝胶的含水率，没有观察到生根。CA在氧气含量为10%的气氛中保存20天后，CA的根再生受到抑制，但当CA转移到氧气为21%的常压下时，CA的根再生得以恢复。提出了一种人工种子，将CA作为细胞包裹体材料，包裹在覆盖有涂层的海藻酸凝胶中。从防止海藻酸凝胶干燥同时保持CA的氧气利用率来保护CA的根再生潜力的角度，测试了乙烯-乙酸乙烯酯-丙烯酸三元共聚物、石蜡和聚有机硅氧烷的不同涂层材料。石蜡具有高效的耐干性和足够的透氧性，因此被选为合适的涂层材料。当海藻酸凝胶中蔗糖浓度大于240molm~(-3)~(-3)时，在25ﾟ℃下与约0.5 mm厚的石蜡层在空气中反应60d，CA的再生率可达90%。本研究提出的人工种子胶囊可用于烟草愈伤组织等植物细胞的保存。

1. The effects of water content in agar gel used as medium and oxygen level in the gas phase on the adventitious root regeneration of cell aggregates (CA) derived from horseradish hairy roots were investigated in cultures at 25ﾟC.The number of roots emerging from CA was highly dependent on water content of the agar gel and no root regeneration was observed at a gel water content of 66% during culture time of 20 days.2. CA root regeneration was suppressed when the CA were kept for 20 days in an atmosphere oxygen composition of 10% but was restored upon transfer of the CA to normal atmosphere of 21% oxygen.3. An artificial seed was proposed using the CA as a cell inclusion material encapsulated in alginate gels covered with coats. From the perspective of conserving the root regeneration potential of the CA by preventing the drying of alginate gel while keeping oxygen availability to the CA,different coat materials of ethylene vinyl acetate acrylic acid terpolymer, paraffin and polyorganosiloxane were tested. paraffin was selected as a suitable coat material because of its efficient drying tolerance and adequate permeability to oxygen.4. The regeneration efficiency of 90% could be obtained from the CA,stored in alginate gel convered with paraffin coat of about 0.5 mm thickness at 25ﾟC for 60 days in air, when sucrose concentration in the gel was over 240 mol m^<-3>.5. The artificial seed capsules proposed in the present study were available for the conservation of plant cells such as tobacco callus.