哈密大枣叶片不定芽再生体系研究

建立哈密大枣叶片离体再生体系,为遗传转化奠定基础.采用哈密大枣叶片为外植体,研究基本培养基、激素浓度、暗培养时间、AgNO3等因素对离体叶片不定芽诱导的影响,并获得再生植株.MS、NN69、WPM三种培养基相比较,NN69更适合做为诱导愈伤组织的基本培养基；TDZ诱导叶片再生不定芽的效果显著优于 BA；再生培养基中添加1 mg/L AgNO3对不定芽的形成有显著的影响（P ＜0．05）；培养初期经过2周避光培养更有利于提高再生效率；采用10 mg/L维生素 C浸泡15 min可以防止褐化,并能提高不定芽再生率；培养基中添加聚乙烯吡咯烷酮（PVP）或者维生素C,不定芽再生率无显著提高（P ＞0．05）；培养基添加活性炭（AC）会抑制外植体的分化.叶片在 NN69＋1．0 mg/L TDZ＋0．20 mg/L IBA＋AgNO31．0 mg/L培养基上,暗培养2周后转入光照培养,出芽外植体转入 MS＋1 mg/L 6GBA＋0．20 mg/L IBA 不定芽再生效果最好.不定芽生长至3 cm 以上时,转至0．40 mg/L IBA的1/2MS培养基上进行诱导生根.

Studies on Adventitious Bud Regeneration System of Zizyphus jujuba Hami Leaves

The obj ective of this research was to establish a highly frequent and efficient regeneration sysG tem of Zizyphus jujube,so as to make preparation for genetic transformation.Using in vitro leaves of Zizyphus jujuba Hami as explants,effects of the basic mediumhormone concentrations,darkincubation time,AgNO3 and other factors on induction rate of adventitious bud from leaves were studied.The results showed,MS,NN69 ,WPM three media were compared with one another,NN69 were more suitable as the basic medium of callus;The efficiency of TDZ was significantly higher than BA in the induction of adventiG tious bud from leaf.AgNO3 addition could greatly increased the regeneration frequencyand the dark culture in the first two week was much more advantageous to increasing the regeneration frequency;The regeneraG tion bud soaked with 10 mg/L vitamin C for 15 min can prevent browing and improve regeneration tate of adventitious bud;medium supplemented with PVP,or vitamin C regeneration bud was not significantly imG proved;medium supplemented with AC inhibits differentiation of explants.The adventitious buds were sucG cessively cultured in MS＋1.0 mg/L 6GBA＋0.20 mg/L IBA until they grew 3 cm long roots and then they were transferred and cultured in the 1/2MS medium to which 0.40 mg/L IBA was added,and this could better induce them to root.