荔枝胚性悬浮细胞系的快速建立及其体胚植株的再生  

Rapid Establishment of Embryogenic Cell Suspensions and Plant Regeneration via Somatic Embryogenesis in Litchi

作　　者：赖钟雄[1] 黄浅[1] 林秀莲[1] 林玉玲[1] 陈义挺[1] 赖呈纯[1] 蔡英卿[1] 

机构地区：[1]福建农林大学园艺植物生物工程研究所,福州350002

出　　处：《中国农学通报》2007年第1期28-32,共5页Chinese Agricultural Science Bulletin

基　　金：国家自然科学基金"通过微原生质体融合将荔枝的部分基因组转移给龙眼"(No.39870544);国家教育部霍英东教育基金会高等学校青年教师基金"利用微核技术将龙眼的部分基因组转移给荔枝"(No.71026);福建省重大科技专项"名特优果树种质资源的保护及应用研究"(2004NZ02-2)资助项目的部分内容。

摘　　要：荔枝幼胚诱导的胚性培养物在低糖条件下连续继代4～6次左右,可筛选到颗粒细小、不含原胚的松散型胚性愈伤组织;以这种松散的胚性愈伤组织作为起始材料,在附加2,4-D2mg/L或2,4-D2mg/L、KT1mg/L、AgNO35mg/L的MS液体启动培养基上振荡培养(100～120r/min)10～14d,即可建立起分散性良好的胚性悬浮细胞系。采用激素减半的2种启动培养基交替继代培养或周期性固体-液体轮回培养,可以长期保持胚性悬浮细胞系。荔枝胚性悬浮细胞在附加NAA0.1mg/L、KT或Ze5mg/L、肌醇100mg/L、蔗糖50g/L、琼脂10g/L的MS固体培养基上诱导体胚,25～40d后可形成大量胚状体,诱导体胚数量达10,000个/gFW以上。经过成熟培养后,正常的体胚75%以上萌发再生完整植株。The friable embryogenic callus with small grains and without proembryo formation was screened in the process of continuous subcultures for 4-6 times on the medium with low content of sucrose from embryogenic cultures induced from immature embryos in litchi （Litchi chinensis Soon.）. The well-scattered embryogenic cell suspensions could be rapidly established from the friable embryogenic callus in 2 liquid initial MS media respectively supplemented with 2,4-D 2 mg/L or 2,4-D 2 rag/L, KT1 mg/L and AgNO3 5 mg/L shaking at 100-120 r/rain within 10-14 days. Long-term maintenance of the embryogenic cell suspensions was achieved via alternative cultures in the 2 revised initial medium only with half-strength of phytohormones or through the method of periodical in-turn liquid-solid culture. The embryogenic＇ suspension cells formed somatic embryos at the frequency of over 10,000 embryoids/g FW on the solid MS medium amended with NAA 0.1 mg/L, KT or Ze 5 rag/L, sucrose 50 g/L, agar 10g/L and myo-inositol 100 mg/L within 25-40 days. Normal somatic embryos were converted into plantlets by over 75% after maturation.

关 键 词：荔枝 胚性悬浮细胞 体胚发生 植株再生 

分 类 号：S667.1[农业科学—果树学]