摘要
以高产优质橡胶品种热研88-13的幼嫩种子内珠被为外植体材料,对愈伤组织诱导培养基和继代培养基中的影响因素如植物生长调节剂的种类和浓度、钙离子浓度等进行了研究。经过连续6个月的继代选择培养,逐渐诱导出易碎的胚性愈伤组织,适宜的易碎胚性愈伤组织诱导培养基为:MH基本成分,添加0.5 mg/L KT,2.0 mg/L 2,4-D,11.0 mmol/L CaCl2,80.0 g/L蔗糖和2.0 g/L Phytagel。组织学切片证明长期继代培养的易碎胚性愈伤组织维持了胚性的状态。取继代培养2 a多的易碎胚性愈伤组织诱导体细胞胚的形成,得到了16 000多个体细胞胚。将这些胚进行进一步的转移培养,得到了115株再生植株。
The inner integument from the young seeds of rubber tree clone Reyan88-13 was used to induce embryogenic callus.The different culture parameters such as the different types and concentrations of plant growth regulators and different calcium concentrations were studied in embryogenic callus induction media and subculture media.The proper culture parameters were determined.Friable and embryogenic callus were gradually obtained via continuous subculture and selection during 6 months.The suitable friable and embryogenic callus induction medium was：MH basal components,supplemented with 0.5 mg/L KT,2.0 mg/L 2,4-D,11.0 mmol/L CaCl2,80.0 g/L sucrose and 2.0 g/L Phytagel.Long-term subculture had been carried on for the friable and embryogenic calli.Histological sections proved that the calli proliferated by long-term subculture maintained the embryogenic characters.The friable and embryogenic calli of rubber tree clone Reyan88-13 that had been subcultured for more than 2 years were used for inducing somatic embryos.More than 16 000 somatic embryos were acquired,then 115 regeneration plantlets were obtained.
出处
《热带作物学报》
CSCD
2010年第12期2166-2173,共8页
Chinese Journal of Tropical Crops
基金
国家自然科学基金项目“橡胶树胚性细胞悬浮培养及其植株再生的研究”(No.30760128)
科技部国际科技合作项目“橡胶树易碎愈伤组织长期培养植株再生和超低温保存研究”(No.2008DFA32020)
海南省重点科技项目“橡胶树花药胚性愈伤组织的超低温保存和冻后植株再生的研究”(No.070105)
行业科技专项子课题“橡胶树胚性细胞悬浮培养植株再生体系建立的研究”(No.nyhyzx07-033-6)
基本科研业务费专项资金项目“橡胶树单细胞遗传转化体系建立的研究”(No.XJSYWFZX2009-01)
农业部“948”项目(No.2010-S7)资助
关键词
橡胶树
易碎胚性愈伤组织
体细胞胚
继代培养
植株再生
Hevea brasiliensis
Friable embryogenic callus
Somatic embryo
Subculture
Plant regeneration
