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

人造种子胶囊中植物细胞的长期保存和有效再生

• 批准号: 08650943
• 负责人: TAYA Masahito
• 金额: $ 1.15万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08650943/
• 关键词: BIOCHEMICAL ENGINEERING; PLANT CELLS; ARTIFICIAL SEED; PARAFEIN THIN FILM; ALGINATE GELS; CELL CONSERVATION

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.

1.在25 ℃培养条件下，研究了琼脂凝胶含水量和气相含氧量对辣根毛状根细胞团（CA）不定根再生的影响。CA不定根的数量与琼脂凝胶含水量密切相关，当凝胶含水量为66%时，无根再生培养时间为20 d. CA根再生在10%的大气氧浓度下保存20天后受到抑制，但转移到21%的正常大气中后恢复.提出了一种人工种子，使用CA作为细胞包含物材料封装在覆盖有涂层的藻酸盐凝胶中。从通过防止藻酸盐凝胶干燥同时保持CA的氧气可用性来保存CA的根再生潜力的角度来看，测试了乙烯乙酸乙烯酯丙烯酸酯、石蜡和聚有机硅氧烷的不同涂层材料。选择石蜡作为合适的涂层材料，因为其有效的干燥耐受性和足够的氧气渗透性。当蔗糖浓度大于240 mol/L时，CA在25 ℃下保存60 d，再生率可达90%<-3>.本研究提出的人工种子囊可用于烟草愈伤组织等植物细胞的保存。

项目成果

馬場 健志: "人工種子保存中の細胞生存活性に及ぼす酸素の効果" 化学工学会姫路大会講演要旨集. 304-305 (1996)

Kenji Baba：“人工种子保存过程中氧气对细胞存活活性的影响”化学工程师学会姬路会议记录 304-305（1996）。

K.Baba: "Conservation of Plant Cells at Room Temperature" Proc.of Annual Meeting of the Soc. of Ferment.Bioeng.Japan. 80 (1996)

K.Baba：“室温下植物细胞的保存”学会年会议程。

P.M.Doran: Hairy Roots-Culture and Applications-. Harwood Academic Publishers, 1-239 (1997)

P.M.Doran：毛根-培养和应用-。

田谷 正仁: "外部環境因子による植物細胞機能の改変とその利用" 化学工学会第63年会講演要旨集. 3巻. 150-151 (1998)

Masahito Taya：“外部环境因素对植物细胞功能的修改及其利用”第 63 届化学工程师学会年会论文集，第 3 卷，150-151（1998 年）。

K.Baba: "Effect of Oxygen on Cell Viability during Conservation of Artificial Seeds" Proc.of Himeji Meeting of the Soc. of Chem.Engrs., Japan. 304-305 (1996)

K.Baba：“人工种子保存过程中氧气对细胞活力的影响”Proc.of Himeji Meeting of the Soc。