胡杨花药再生体系的建立

Establishment of Anther Regeneration System of Populus euphratica

以胡杨花药为外植体,消毒灭菌后诱导愈伤,使用正交设计探究了生长素和分裂素在愈伤诱导中的作用,继而诱导愈伤分化,促使不定芽生根,最终建立了高效地胡杨花药再生体系。结果表明:70%乙醇消毒30 s后,8%的次氯酸钠消毒2 min,无菌水清洗3遍为最佳消毒方法;H培养基+1.0 mg/L NAA+1.0 mg/L BA+5.5 mg/L琼脂+30 mg/L蔗糖为最佳愈伤诱导培养基,诱导率达89%;MS培养基+1.0 mg/L NAA+0.2 mg/L BA+5.5 mg/L琼脂+30 mg/L蔗糖为最佳分化诱导培养基,分化率约50%;添加了1.2 mg/L IBA+5.5 mg/L琼脂+20 mg/L蔗糖的1/2MS培养基中分化芽的生根率最高,可达96.7%。

In the present study,anthers of Populus euphratica were used as explants to induce callus after disinfection and sterilization.The orthogonal design was used to explore the role of auxin and cytokinin in callus induction,followed by induction of callus differentiation and multiple shoots rooting.Finally,an efficient regeneration system of P.euphratica was established.The results show that the best method for sterilization as following:70%alcohol soak for 30 seconds,then 8%sodium hypochlorite for 2 minutes,finally sterile water for 3 times;H medium supplemented with 1.0 mg/L NAA and 1.0 mg/L BA+5.5 mg/L agar+30 mg/L sucrose induced the highest rate of callus formation,the induction rate was 89%;MS medium+1.0 mg/L NAA+0.2 mg/L BA+5.5 mg/L agar+30 mg/L sucrose as the best medium for differentiation induction,the differentiation rate is about50%;the rooting rate was the highest in 1/2 MS medium supplemented with 1.2 mg/L IBA+5.5 mg/L agar+20 mg/L sucrose,it can be reach 96.7%.