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以红刺玫无菌苗为初始材料,通过探讨外植体、培养条件,以及激素配比浓度和暗培养时间对愈伤组织分化芽的影响,建立了红刺玫叶片愈伤组织诱导的再生体系:MS为初始培养基,暗培养21 d,愈伤组织诱导率达到100%。分化培养基为MS+TDZ 1.5 mg/L+NAA0.05 mg/L,暗培养8 d,芽分化率达48%;通过遗传转化条件优化,建立了以红刺玫愈伤组织为转化受体,通过根癌农杆菌介导,以红刺玫的DFR-RNAi为表达载体,GUS为标记基因的遗传转化体系,转化效率达到50%。
Asexual red rose seedling was used as the initial material to study the effect of explants, culture conditions, hormone concentration and dark culture time on callus bud differentiation, System: MS as the initial medium, dark culture 21 d, callus induction rate of 100%. Differentiation medium was MS + TDZ 1.5 mg / L + NAA 0.05 mg / L, dark culture 8 d, bud differentiation rate of 48%; through the optimization of genetic transformation, established a red rose callus as the transformation receptor , Which was mediated by Agrobacterium tumefaciens. Its transformation efficiency was 50% with the genetic transformation system of GUS as the marker gene by using DFR-RNAi as the expression vector.