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研究了植物激素、刻伤、放置方式等因子对广柑子叶和上胚轴切段离体再生的影响 ,建立了一个高效的再生系统 .该系统的最佳条件是 :采用实生苗子叶和上胚轴茎段作为外植体 ,MS无机盐 +B5维生素 +5%蔗糖 +0 .8%琼脂 +7.5mg/ L BA+1mg/ L NAA(p H5.7)作为不定梢诱导培养基 ;子叶沿中间纵切为二 ,同时在非切面部分横向刻伤 ,切面朝下与培养基表面接触放置 ;上胚轴则切成 0 .5～0 .8cm长的切段 ,横放于培养基上 .培养条件为恒温 (2 6± 1)℃ ,首先在黑暗下培养 15d,而后转移至16 / 8h光 /暗周期下 ,共经过 70 d培养 ,95%子叶外植体均能发生不定梢 ,平均每个外植体上发生不定梢数为 11.11个 ,96 %的上胚轴切段能发生不定梢 ,平均每个外植体上发生不定梢数为 5个 .这一系统再生频率高 ,操作简便 ,适用于广柑的遗传转化
The effects of plant hormones, engravings, and placement methods on in vitro regeneration of cuttings and hypocotyl cuttings of C. giganteum were studied, and an efficient regeneration system was established.The optimum conditions of the system were as follows: The stems of the hypocotyls were used as explants, MS inorganic salts + B5 vitamins + 5% sucrose +0.8% agar + 7.5 mg / L BA + 1 mg / L NAA (p H5.7) as adventitious shoot induction medium; Along the middle of longitudinal slitting two, at the same time in the non-section part of the lateral engraved, face down with the medium surface contact placed; epicotyls cut into 0.5 ~ 0.8cm long cut, The culture conditions were constant temperature (26 ± 1) ℃, cultured in the dark for 15 days and then transferred to the 16/8 h light / dark cycle. After 70 days of culture, 95% of cotyledon explants could not shoot indefinitely , An average of 11.11 indefinite shoots per explant occurred, 96% of the hypocotyl cuttings can occur indefinite tip, the average number of indefinite shoots per explant is 5. The system of high frequency of regeneration , Easy to operate, suitable for genetic transformation of Citrus grandis