‘小白杏’诱导愈伤组织及不定芽体系的优化

System Optimization for Inducing Callus and Buds from 'Xiaobaixing' ( Prunus armeniaca)

为建立快速繁殖离体培养再生体系并为遗传转化奠定基础,本研究以‘小白杏’胚和叶片为材料,探讨了氯化汞和次氯酸钠消毒的方法和时间对种子和种胚的影响,2,4-D(0.5,1.0,1.5 mg/L)、ZT(0.5,1.0,1.5 mg/L)和TDZ(1.0,2.0,3.0 mg/L)浓度配比对‘小白杏’胚诱导愈伤组织再分化不定芽的作用,及2,4-D(0.5,1.0 mg/L)、IBA(0.5,1.0 mg/L)、NAA(0.5,1.0 mg/L)浓度配比对‘小白杏’叶片诱导愈伤组织再分化不定芽的作用。结果表明:使用10%的次氯酸钠消毒10 min,种胚污染率和死亡率最低,分别为2.6%、0.7%,存活率最高,为96.7%;在3/4MS+2.0 mg/L TDZ+1.0 mg/L 2,4-D处理下,胚诱导愈伤组织形成率和不定芽诱导率最高,分别为98%、96%;在3/4MS+2.0 mg/L AgNO_3+1.0 mg/L ZT+2.0 mg/L TDZ+1.0mg/L IBA处理下,叶片诱导愈伤组织形成率和不定芽诱导率最高,分别为96%、93%。综合来看,胚诱导愈伤组织再分化形成不定芽的诱导率优于叶片,种子和种胚消毒的最佳方案是10%的次氯酸钠消毒10 min;胚诱导愈伤组织再分化形成不定芽最佳配方是3/4MS+2.0 mg/L TDZ+1.0 mg/L 2,4-D;叶片诱导愈伤组织再分化形成不定芽的最佳配方是3/4MS+2.0 mg/L AgNO_3+1.0 mg/L ZT+2.0 mg/L TDZ+1.0 mg/L IBA。

To establish rapid propagation and regeneration system for ‘Xiaobaixing'in vitro and provide foundations for genetic transformation,with embryo and leaf as explants,we studied the effects of disinfection with mercuric chloride and sodium hypochlorite on seed and embryo,the effects of different concentration ratios of 2,4-D( 0. 5,1. 0,1. 5 mg/L),ZT( 0. 5,1. 0,1. 5 mg/L) and TDZ( 1. 0,2. 0,3. 0 mg/L) on embryo induction,and that of 2,4-D( 0. 5,1. 0 mg/L),IBA( 0. 5,1. 0 mg/L) and NAA( 0. 5,1. 0 mg/L) on leaf induction. The results showed that the pollution rate and mortality were the lowest with 10% sodium hypochlorite disinfecting seed and embryo for 10 minutes,which were 2. 6% and 0. 7% respectively,and the survival rate was the highest as 96. 7%. In the treatment condition of 3/4 MS containing with 2. 0 mg/L TDZ and 1. 0 mg/L 2,4-D,the rate of callus formation and adventitious bud induction with embryo as explant were the highest,which were 98% and 96%,respectively. In the treatment condition of 3/4 MS containing with 2. 0 mg/L AgNO_3,1. 0 mg/L ZT,2. 0 mg/L TDZ and 1. 0 mg/L IBA,the rate of callus formation and adventitious bud induction with leaf as explant were the highest,which were 96% and 93% respectively. In conclusion,the cal-lus differentiation to adventitious buds with embryo as explant was better than that with leaf as explant. The best treatment for disinfecting seed and embryo was both with 10% sodium hypochlorite for 10 minutes. The optimal medium for inducing callus redifferentiation to adventitious buds of embryo was 3/4 MS containing with 2. 0 mg/L TDZ and 1. 0 mg/L 2,4-D,while that for leaves was 3/4 MS containing with 2. 0 mg/L AgNO_3,1. 0 mg/L ZT,2. 0 mg/L TDZ and 1. 0 mg/L IBA.