桃砧木GF677叶片愈伤组织诱导和保存

Studies on Callus Induction and Conservation from Leaves of Peach Rootstock GF677

[目的]研究桃砧木GF677(Prunus amygdalus×Prunus persica)叶片愈伤组织的诱导和保存技术。[方法]以桃砧木GF677试管苗为材料,研究激素含量、基本培养基、AgNO3、暗培养时间、碳源等因素对其叶片愈伤组织诱导和保存的影响。[结果]选取继代时间为28 d左右的试管苗幼嫩叶片,暗培养21 d后转移至光培养进行愈伤组织诱导,较佳的培养基配方为LP基本培养基+TDZ 1.0 mg/L+6-BA 0.6 mg/L+2,4-D 0.2 mg/L+AgNO30.5 mg/L,添加山梨醇20.0 g/L,琼脂6.0 g/L;选取继代时间为21 d左右的愈伤组织暗培养保存,保存较佳的培养基配方为LP基本培养基+6-BA 0.5 mg/L+2,4-D 0.5 mg/L+CH 0.2 g/L,添加蔗糖30.0 g/L,琼脂6.0 g/L。[结论]为桃砧木GF677的遗传转化和无性系变异筛选等工作奠定了基础。

[Objective] To study the callus induction and conservation from leaves of peach rootstock GF677（Prunus amygdalus × Prunus persica）.[Method] Experiment was conducted with the leaves of peach rootstock GF677 in vitro for studying the effects of hormone dose,basic medium,AgNO3,dark culture time and carbon sources on callus induction and conservation.[Result] The young leaves along with their petioles from 28-day-old in vitro grown shoots were used as explants,which were transferred to light culture after 21 d dark treatment.The result showed that the optimal medium for callus induction was LP ＋1.0 mg/L TDZ ＋ 0.6 mg/L 6-BA ＋0.2 mg/L 2,4-D ＋ 0.5 mg/L AgNO3 ＋ 20.0 g/L sorbitol ＋ 6.0 g/L agar.The optimal medium for callus conservation was LP ＋ 0.5 mg/L 6-BA ＋ 0.5 mg/L 2,4-D ＋ 0.2 g/L CH ＋30.0 g/L sucrose ＋ 6.0 g/L agar.The callus was in darkness all along and subcultured for every 21 d.[Conclusion] The study laid a foundation for the genetic transformation and somaclonal variation of peach rootstock GF677.