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以银中杨三倍体无性系带芽茎段为外植体,通过芽生芽途径建立了三倍体无性系组织培养转基因再生系统。筛选出不同阶段最佳培养基与激素浓度分别为:(1)初代培养阶段:1/2MS+TDZ0.02mg/L+NAA0.05mg/L,外植体接种10d后腋芽萌发,30d后腋芽的萌发率可达96%,芽平均长度为6.0cm;(2)继代培养阶段:MS+BA0.05mg/L+NAA0.02mg/L,接种30d后分化率可达100%,茎高为3.5cm,60d后茎高达7.0cm;(3)生根培养阶段:1/2MS+NAA0.05mg/L+IBA0.20mg/L,20d后生根率可达100%。
The triploid clonal tissue culture transgenic regeneration system was established by means of budding and bud pathway with the stem segments of buds of triploid inbred lines of Yinzhong as explants. The best culture medium and hormone concentration in different stages were screened out as follows: (1) primary culture stage: 1 / 2MS + TDZ0.02mg / L + NAA0.05mg / L, explants 10d after inoculation axillary bud germination, 30d axillary bud (2) The subculture stage: MS + BA0.05mg / L + NAA0.02mg / L, the rate of differentiation up to 100% 30 days after inoculation, the stem height is 3.5 cm, and the height of the stem was up to 7.0 cm after 60 days. (3) The rooting stage: 1 / 2MS + NAA0.05mg / L + IBA0.20mg / L.