甘蔗胚性愈伤高效诱导系列研究Ⅰ.幼叶离体培养直接诱导胚性愈伤发生

A Serial Studies on High-efficiency Induction for Sugarcane Embryonic Callus Ⅰ. Vitro Direct Induction of Embryonic Callus in Sugarcane Leaf Explants

以ROC22心叶为外植体,研究了取材部位、培养基蔗糖含量等因素对甘蔗胚性愈伤发生的影响,重点探讨了不同植物激素和生长调节剂组合的作用,建立了无需继代、直接诱导高质量甘蔗胚性愈伤发生的体系。结果表明：顶端生长点以上5-6 cm区段最内层3片心叶为最适外植体;MS＋2,4-D 3.0 mg/L＋Dicamba 1.0 mg/L＋6-BA 0.1 mg/L＋DA-6 6.0 mg/L＋CSN 4.0 mg/L＋蔗糖40 g/L为最佳培养基;经28 d暗培养,心叶无需继代即可分化成为胚性愈伤,诱导率可达42.67%。所得胚性愈伤质量高、分化强,可用于甘蔗遗传转化研究。

Direct and subculture-free induction system for high-quality embryonic callus of sugarcane was established with furled young leaves of ROC22 sectioned as explants, and effects of different parts of young leaves, sucrose content of medium, especially the combination of various phytohormone and growth regulators to induction rate were investigated. The results showed that longitudinal region between 5 to 6 cm above the apical meristem of inner 3 leaves was optimal as explant; MS medium with 2,4-D 3.0 mg/L＋ Dicamba 1.0 mg/L ＋ 6-BA 0.1 mg/L ＋ DA-6 6.0 mg/L ＋ CSN 4.0 mg/L ＋ Sucrose 40 g/L was screened to be as the optimum. After treating 28 d of darkness and non-subculture, the explants could be directly inducted into embryonic callus rate up to 42.67%. Thus the embryonic callus of high quality and strong ability of regeneration from the subculture-free induction system was particularly suitable for sugarcane gene-transfer.