日本晴水稻悬浮细胞系的建立和保存

本研究旨在建立日本晴水稻悬浮细胞系和保存悬浮细胞。在添加2 mg.L-1 2,4-D的NB培养基之后日本晴的成熟胚被诱导愈伤组织形成,2个月的继代后,愈伤组织在2 mg.L-1 2,4-D的NB液体培养基上用于悬浮细胞的培养。3个月的继代后,得到理想的悬浮细胞。关于培养的植物细胞的超低温保存程序大多数报道是两步保存法,采用含有脱落酸激素的预培养溶液培养,同时利用化学冷冻保护剂处理,并在-30℃下放置2 h后投入液氮保存,复苏后细胞的存活率(氯化三苯四氮唑还原法)为80%以上。

Establishment and cryopreservation of rice suspension cells line

The article means to establish rice suspension cells line and conservated suspension cell.Rice mature-embryo were induced for calli formation on NB solid medium containing 2 mg·L-1 2,4-D,after subcultured for about two months,calli were used for cells suspension culture in NB liquid medium containing 2 mg·L-1 2,4-D.After subcultured for three months,ideal suspension cells were got.Most reported cryopreser-vation protocols for plant cells cultures for follow a two-step procedure.At first,pre-culture with pre-solution containing ABA,then treatment with chemical cryoprotectants,and combine with stored at-30 ℃ for two hours before quenching in LN for storage.Activity of cells(TTC) above 80% after thawing.