簸箕柳组培再生体系的建立

Plant regeneration of Salix suchowensis through tissue culture

【目的】建立簸箕柳组培再生体系,为在簸箕柳中开展功能基因组研究奠定基础。【方法】分别以簸箕柳带腋芽茎段和种子为外植体,比较不同消毒方法、基本培养基类型、光照条件及植物生长调节剂等因素对不同外植体脱分化和再分化能力的影响。【结果】簸箕柳组培基本培养基成分以MS+蔗糖(30 g/L)+Phytagel (植物凝胶)(4.4 g/L)为宜。以幼嫩茎段为外植体,最佳消毒方法为70%(体积分数)酒精处理30 s+20%(质量分数)的次氯酸钠处理10 min,适宜诱导腋芽的生长调节剂组合为6-BA(1 mg/L)+NAA (0.01 mg/L),丛生芽继代增殖的适宜生长调节剂配比为6-BA (0.5 mg/L)+NAA (0.05 mg/L),适宜腋芽生根的生长调节剂组合为IBA (0.2 mg/L)+NAA (0.03 mg/L)。以种子为外植体,最佳消毒方法为70%(体积分数)酒精处理10 s+20%(质量分数)的次氯酸钠处理2 min,光照条件下诱导愈伤组织的生长调节剂配比为6-BA (0.5 mg/L)+NAA (0.3 mg/L),并利用6-BA (0.5 mg/L)+NAA (0.5 mg/L)和6-BA (0.5 mg/L)+NAA (0.8 mg/L)两种组合方式交替使用进行愈伤组织的继代培养;愈伤组织分化成芽的生长调节剂配比为6-BA (0.5 mg/L)+TDZ (0.2 mg/L),生根培养基只需添加NAA (0.01 mg/L)。【结论】以茎段和种子作为外植体,通过不同的再生途径,均可以建立簸箕柳组织培养再生体系。

【Objective】In this study,we aimed to establish an efficient plant regeneration protocol for Salix suchowensis through tissue culture,which would serve as a foundation for developing a genetic transformation system for this woody shrub.【Method】By using young stems with axillary buds and mature seeds as explants,the influences of different factors on plantlet regeneration were systematically studied,including sterilization methods,types of the basic medium,light conditions,and plant growth regulators.【Result】The optimal basic medium for S.suchowensis was Murashige and Skoog(MS)medium+sucrose(30 g/L)+Phytagel(4.4 g/L).When using young stem nodal segments as explants,the best sterilization method was 70%ethanol for 30 sec and then 20%sodium hypochlorite for 10 min,while the optimal medium for axillary buds induction and multiplication was the basic medium supplemented with 6-BA(1 mg/L)+NAA(0.01 mg/L)and 6-BA(0.5 mg/L)+NAA(0.05 mg/L),respectively,and the appropriate medium for rooting was the basic medium+IBA(0.2 mg/L)+NAA(0.03 mg/L).While for mature seeds,the optimal sterilizing treatment was 70%ethanol for 10 sec and then 20%sodium hypochlorite for 2 min;the appropriate medium for callus indution,proliferation and subsequent shoot differentiation was the basic medium supplemented with 6-BA(0.5 mg/L)+NAA(0.3 mg/L),6-BA(0.5 mg/L)+NAA(0.5 or 0.8 mg/L),6-BA(0.5 mg/L)+TDZ(0.2 mg/L),respectively,and the best rooting medium was the basic medium containing NAA(0.01 mg/L).【Conclusion】We established a regeneration protocol for S.suchowensis from the axillary buds or callus by using young stem nodal segments and mature seeds as explants respectively.