尾巨桉M8无性系增殖培养技术研究

Study on the Multiplication and Culture Technology of Eucalyptus urophylla×E.grandis M8 Clones

选用已经通过无性系测定的优良无性系单株尾巨桉(Eucalyptus urophylla×E.grandis)M8,以其半木质化嫩枝为外植体,采用组培繁殖中遗传稳定性高的丛芽发生途径,探讨基本培养基、6-BA、IBA、蔗糖和pH这5个因子分别对增殖效果的影响。结果表明:(1)采用MS+6-BA 0.2 mg/L+蔗糖30 g/L+卡拉胶6.8 g/L,pH为5.8的培养基进行尾巨桉M8外植体腋芽的诱导时,10~15 d为出芽高峰期,平均污染率为15.3%,平均诱导率为77.9%;(2)适宜尾巨桉M8增殖的最佳培养基为改良MS培养基、6-BA浓度为0.2~0.5 mg/L、IBA浓度为0.05~0.2 mg/L、蔗糖浓度为30 g/L、pH值范围为5.4~6.0,继代周期为20 d,增殖系数可达4.49,有效芽数可达30棵/瓶。因此,尾巨桉M8较适宜的诱导培养基为:MS+6-BA 0.2 mg/L+蔗糖30 g/L+卡拉胶6.8 g/L,可实现腋芽萌发快,污染率少,诱导率高。最优的增殖培养基为改良MS+6-BA 0.2 mg/L+IBA 0.1 mg/L+蔗糖30 g/L+卡拉胶6.8 g/L,pH为5.4~6.0,可实现继代苗叶片舒展,生长健壮,提高组培效率,发挥组培快繁的优势。

In order to investigate the effects of minimal medium,6-BA,IBA,sucrose and pH on the multiplication of Eucalyptus urophylla×E.grandis M8 clones,an excellent clonal single plant,and its semiligniated shoots were selected as explants.The results showed that:(1)when the medium of MS+6-BA 0.2 mg/L+sucrose 30 g/L+carrageenan 6.8 g/L with pH 5.8 was used to induce the axillary buds of M8 explants,the germination peak was observed at 10-15 days,with an average pollution rate of 15.3%and an average induction rate of 77.9%.(2)the optimal medium for M8 multiplication was the improved MS medium,with a concentration of 0.2-0.5 mg/L for 6-BA,0.05-0.2 mg/L for IBA,30 g/L for sucrose,a pH range of 5.4-6.0,a subculture period of 20 days,which could get a multiplication coefficient of 4.49,and an effective number of 30 shoots per bottle.Therefore,MS+6-BA 0.2 mg/L+sucrose 30 g/L+carrageenan 6.8 g/L is the appropriate induction medium for M8,which can realize rapid axillary bud germination,low pollution rate and high induction rate.The optimal multiplication medium is improved MS+6-BA 0.2 mg/L+IBA 0.1 mg/L+sucrose 30 g/L+carrageenan 6.8 g/L with a pH of 5.4-6.0,which could realize the extension of seedling leaves and robust growth,improve tissue culture efficiency and give full play to the advantages of tissue culture and rapid propagation.