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本文以草莓主栽品种‘早红’组培苗离体叶片和叶柄为外植体,进行叶龄、暗培养、植物生长调节剂配比及抗生素敏感性研究,建立草莓高效遗传转化的受体系统。在含3.0mg/L6-BA与0.1mg/L2,4-D的MS培养基上,30d叶龄的叶片再生频率高达98.31%,平均每叶片再生芽数5.09个,叶柄切段的再生频率为89.25%,平均每叶柄切段再生芽数4.92个,叶片的再生频率略高于叶柄;不定芽在含0.2mg/L6-BA与0.2mg/LGA3的MS继代培养基上培养成苗。将生长状态良好的不定芽转至含0.2mg/LIBA的1/2MS培养基上生根,生根率达100%,平均生根数量16.27条,平均根长1.85cm。抗生素敏感性试验表明,草莓外植体适宜的卡那霉素选择压力为25mg/L,头孢霉素的筛选浓度为300mg/L。本研究建立的再生体系可作为草莓遗传转化的受体系统。
In this study, leaves and petioles of the cultivated early-red strawberry seedlings were used as explants to study leaf age, darkness, ratio of plant growth regulators and antibiotic sensitivity, and to establish a high efficient genetic transformation receptor for strawberry system. In MS medium containing 3.0mg / L 6-BA and 0.1mg / L 2,4-D, the leaf regeneration frequency was as high as 98.31% at leaf age of 30d, the number of regenerated buds per leaf was 5.09. The frequency of regeneration of petiole was 89.25%. The number of regenerated buds per cutting segment was 4.92, and the regeneration frequency of leaves was slightly higher than that of petiole. Adventitious buds were cultured on MS subculture medium containing 0.2 mg / L 6-BA and 0.2 mg / L GA3. The adventitious buds with good growth conditions were transferred to 1/2 MS medium containing 0.2 mg / L IBA, the rooting rate was 100%, the average rooting number was 16.27 and the average root length was 1.85 cm. Antibiotic susceptibility tests showed that the suitable selection pressure of kanamycin for strawberry explants was 25 mg / L and the screening concentration of cefotaxime was 300 mg / L. The regeneration system established in this study can be used as a receptor system for strawberry genetic transformation.