以黑莓品种‘Kiowa’无菌苗叶柄为外植体,研究了培养基类型、暗培养时间、着生位置、激素种类配比和生根条件等对叶柄不定芽诱导的影响,建立了叶柄直接再生体系。结果表明,把‘Kiowa’试管苗继代培养30~40 d后,取自上而下第1~2叶位的叶柄为外植体,接种在MS或1/2MS上,暗培养14 d或21 d叶柄的再生效果较好。‘Kiowa’叶柄不定芽再生的适宜培养基为MS+CPPU 1.0 mg/L+IAA 0.5 mg/L,不定芽再生率达87.46%,平均叶柄再生芽数目达4.42。芽增殖以MS+6-BA 0.5 mg/L+IAA 0.1 mg/L效果最好,增殖系数达5.19;再生植株生根以1/2MS+NAA 0.03 mg/L最佳,生根率达91.67%,平均生根条数为4.00。
Taking the petiole excised from the axenic seedlings of blackberry(Rubus hybrid) cv.'Kiowa' as explants,a high-frequency in vitro direct regeneration system has been developed.The petioles from 30~40 days' mother cultures on MS medium were used as the initial explants to investigate the influences of medium type,dark treatment time,petiole position,exogenous hormones,and rooting conditions on the plant regeneration.The results showed that: after 30~40 days' subculture of 'Kiowa' test-tube seedlings,culturing the petioles from the top first to the second leaf-position on 1/2MS or MS medium for 14 days or 21 days under dark condition could obtain better regeneration effect.The optimal medium for the inducement of adventitious buds from petioles was MS+CPPU 1.0 mg/L+IAA 0.5 mg/L,and the average regenerated bud number reached 4.42 with the regeneration rate of 87.46%.MS+6-BA 0.5 mg/L+IAA 0.1 mg/L was the optimum medium for bud multiplication with the coefficient of 5.19.The optimum rooting medium for petioles was 1/2MS+NAA 0.03 mg/L,in which the rooting rate reached 91.67% and the average number of roots of each plant was 4.00.