摘要
以可可(TheobromaCacaoL.)的子叶为外植体,通过体胚发生途径,诱导再生可可植株。各培养阶段的优化培养基和培养条件为:(1)愈伤组织诱导培养基(PCG):改良DKW+2,4-D3.0mg/L+KT1.0mg/L+TDZ0.01mg/L,在(28+2)℃(以下培养温度均同)温度条件下,暗培养20d,诱导率为96.67%;(2)愈伤组织增殖培养基(SCG):改良DKW+2,4-D3.0mg/L+KT1.0mg/L,暗培养20d;(3)胚状体诱导培养基(ED):改良DKW+Sucrose30g/L,暗培养60￣150d,胚状体诱导产生并发育成熟,胚状体的诱导率为33.33%;(4)成熟胚诱导成苗:①PEC培养基为:改良DKW+Glucose20g/L+Sucrose10g/L,光照为16h/d,培养60d;②采用RD培养基:改良DKW+IBA1.5mg/L+IAA0.5mg/L,光照为16h/d,培养30￣90d后,可得到完整的植株,再生植株的诱导率为42%。
Plants were regenerated through somatic embryogenesis using cotyledon as explants. The optimized medium and condition for each stage were tested as follows, （1） Primary calli medium （PCG） was the improved DKW with 3.0 mg/L of 2,4-D, 1.0 mg/L of KT and 0.01 mg/L of TDZ, explants were cultured on PCG medium in the dark at （28±2）℃ for 20 days. （2） Calli propagation medium （SCG） was the improved DKW with 3.0 mg/L of 2, 4-D, 1.0 mg/L of KT, the calli were cultured on SCG medium for 30 days in the dark. （3） The calli were subcuhured on ED medium with 30 g/L of sucrose, 1 g/L of glucose in the dark for 60 to 150 days, and embryos at different developmental stages （globular, beart torpedo and mature） were present on individual explants at the same time. （4） The mature embryos were transferred to fresh PEC medium every 30 days until shoots with one or two leaves developed. The contents of PEC medium were the improved DKW with 20 g/L of glucose, 10 g/L of sucrose. Shoot-producing embryos with 2 leaves were transferred onto RD medium that was the improved DKW supplemented with 1.5 mg/L of IBA and 0.5 mg/L of IAA. The cultures were maintained under light for 16 h/d. After 30 to 90 days, regenerated plants were obtained.
出处
《热带作物学报》
CSCD
2005年第4期15-19,共5页
Chinese Journal of Tropical Crops
基金
海南省教育厅高等学校科研资助项目(Hjkj200117)。
关键词
可可
子叶
组织培养
再生植株
Theobroma Cacao L. somatic embryogenesis tissue culture plant regeneration
