欧洲丁香品种‘Downfield’花序和花序轴愈伤组织诱导和悬浮培养

Callus Induction and Suspension Culture with Inflorescence and Inflorescence Axis of Syringa vulgaris ‘Downfield’

以欧洲丁香(Syringa vulgaris)花序和花序轴作为外植体,研究基本培养基类型、植物激素类型和质量浓度对欧洲丁香愈伤组织诱导的影响,并通过正交试验研究锥形瓶容积、初始液体培养基体积、旧培养基保留比例、接种质量及抗氧化剂质量浓度对悬浮培养愈伤组织增殖效果的影响。结果表明:最优花序轴愈伤组织诱导培养基为改良型Blaydes+苯基噻二唑基脲(TDZ)0.10 mg/L+2,4-二氯苯氧乙酸(2,4-D)1 mg/L,最优诱导花序愈伤组织的培养基为MS培养基+TDZ 0.10 mg/L+2,4-D 1 mg/L,诱导率均为100%。2,4-D质量浓度显著影响花序和花序轴愈伤组织诱导率(p<0.05),TDZ与2,4-D联合作用显著影响花序轴愈伤组织诱导率(p<0.05)。悬浮培养的最优组合为锥形瓶容积250 mL、初始液体培养基体积150 mL、旧培养基保留1/4、接种质量7.0 g,聚乙烯吡咯烷酮(PVP)质量浓度100 mg/L,维生素C(Vc)质量浓度75 mg/L。首次以欧洲丁香花序和花序轴为材料建立了愈伤组织诱导和细胞悬浮培养体系,为后续开展工厂化育苗和遗传转化提供参考。

The inflorescences and inflorescence axial of Syringa vulgaris as explants were used to study the effects of explant types, basic culture medium types, plant hormone types and concentration to the callus induction rate of S. vulgaris, and design orthogonal experiment was used to explore the effects of erlenmeyerflask volume, initial culture medium volume, retention of old medium, inoculum quality and the content of antioxidants to the callus proliferaton result of suspension culture. The optimal callus induction of inflorescence axis medium was modified as Blaydes+TDZ 0.10 mg/L+2,4-D 1 mg/L, the optimal inflorescence induction medium was MS+TDZ 0.10 mg/L+2,4-D 1 mg/L, the induction rate was up to 100%. The concentration of 2,4-D had significant effect the callus rate of inflorescence and inflorescence axis(p<0.05). The combination of TDZ and 2,4-D significantly affected the inflorescence axial callus rate(p<0.05). The optimal combination of suspension culture was using the volume of 250 mL erlenmeyer flask, and supplemented with 150 mL of initial culture mediumaccompanied by 4.5 g of calli, andafter 15 d, retained of 1/4 old medium for next stage, initial inoculumed 7 g, PVP concentration 100 mg/L, Vc concentration 75 mg/L. This is the first time to report callus induction and start cell suspension culture of inflorescence and inflorescence axis of S. vulgariswith successful results, which would be used for factory breeding and genetic transformation in the future.