【作者】 姜玉东；

【导师】 陈典；

【作者基本信息】 东北农业大学 ， 蔬菜学， 2004， 硕士

【摘要】 以茎尖为外植体，探索分蘖洋葱的组培技术，在此基础上，采用由外植体诱导形成试管苗，再诱导试管鳞茎的技术途径，从培养条件入手，探索试管鳞茎微繁技术，以解决传统接口技术上的弊端，为无毒种苗试管鳞茎的工厂化生产提供理论依据和关键技术。结论如下：1.试管苗的组培技术（1）MS+NAA0.1mg/L+BA0.7mg/L和MS+NAA0.1mg/L+2iP0.7mg/L培养基都适合诱芽培养。 （2）MS+IBA2.5 mg/L和1/2MS+IBA1.5 mg/L +NAA0.05mg/L培养基都适合生根培养。（3）提高C/N比值有利于生根培养。（4）较低浓度的IBA或NAA有利于生根培养，IBA的生根效果优于NAA。2.试管鳞茎的微繁技术（1）糖源试管苗在以蔗糖（化学纯及分析纯）、葡萄糖或绵白糖为糖源的培养基上都能正常生长，果糖不适合生长，可溶性淀粉不能用作糖源；试管苗在以蔗糖（化学纯及分析纯）、葡萄糖、果糖或绵白糖为糖源的培养基上均能形成鳞茎，以分析纯蔗糖的效果最好，鳞茎发生数和鲜重分别达到39.67个和64.93mg，化学纯蔗糖其次，绵白糖居中，葡萄糖和果糖效果最差；绵白糖也是试管苗生长及鳞茎形成的良好糖源，可作为蔗糖的替代品。（2）糖浓度较高的糖浓度抑制试管苗生长，试管苗难于抽叶生根及吸收水分；在一定范围内，提高糖浓度促进鳞茎形成。糖浓度越高，鳞茎形成越早。但浓度过高（150g/L）会削弱形成效果；120g/L的蔗糖浓度最适合鳞茎化培养，鳞茎平均发生数和形成率分别达到31.33个和90.00%。（3）植物激素在无激素或BA和（或）NAA浓度较低时有利于鳞茎形成；NAA1.0mg/L最适合鳞茎化培养，鳞茎直径和鲜重分别达到4.02mm和115.73mg。（4）多效唑在一定范围内，提高多效唑浓度有利于鳞茎形成，但浓度过高（10.0mg/L）会削弱形成效果；添加5.0mg/L多效唑最适合鳞茎化培养，鳞茎发生数、形成率、直径、鲜重和指数分别达到41.00个、92.86%、11.55mm、709.06mg和1.10；多效唑具有很强的矮化和抑制生长效果，一定范围内，效果随浓度的升高而加强，有鳞茎指数<1的情况出现，鳞茎从长圆渐变成扁圆形；多效唑具有延缓生长的效果，鳞茎形成过程约需150d以上；在添加多效唑的培养基上形成的鳞茎呈淡绿色，紫红色外皮较少。（5）烯效唑一定范围内，提高烯效唑浓度有利于鳞茎形成；添加1.0mg/L烯效唑最适合鳞茎化培养，鳞茎发生数、形成率、直径和指数分别达到45.67个、93.34%、8.52mm和1.21；烯效唑具有强烈的促进生根效果，试管苗根系繁密；烯效唑具有较强的矮化效果，一定范围内，效果随浓度的升高而加强，有鳞茎指数<1的情况出现，鳞茎由长圆渐变成圆形；烯效唑具有延缓生长效果，鳞茎形成过程约需150～210d以上；在添加烯效唑的培养基上形成的鳞茎呈淡绿色，紫红色外皮较少。（6）活性炭 <WP=9>活性炭具有很强的促进生长和生根效果；活性炭具有较强的促进鳞茎形成效果，在1～5g/L范围内，提高活性炭浓度有利于鳞茎形成，各处理鳞茎形成率均高达90%以上，其中添加3～5g/L的鳞茎形成率高达100%，但添加5g/L会轻度削弱鳞茎形成效果；添加4～5g/L活性炭最适合鳞茎化培养，鳞茎发生数、直径、鲜重和指数分别达到44.33个、7.82mm、656.91mg和1.68；在添加活性炭的培养基上形成的鳞茎较长，鳞茎指数较大，具有紫红色外皮。3.试管鳞茎的播种栽培将试管鳞茎按直径大小分成A：0～3mm、B：3～6mm、C：6～9mm和D：9～12mm四个等级，经播种栽培后成活率分别为63.33%，86.67%、100%和100%；萌发苗长势随直径的增大而增强。更多还原

【Abstract】 It regarded stem tip of Tillered-onion as explants，on the basis of the technical exploration of tissue culture in Tillered-onion，it adopted the pathway which induced test-tube seedlings from explants then induced bulblets，after this，it explored the technique of micropropagation which proceeded from the condition of tissue culture，in order to solve the malady in traditional technique of interface，and provided factory production of bulblets from virus-free seedlings，the conclusions were as follows：1 the technique of tissue culture for test-tube seedlings（1）The mediums of MS+NAA0.1mg/L+BA0.7mg/ L and MS+NAA0.1mg/L+2iP0.7mg/L were both fit for the culture of induction.（2）The mediums of MS+IBA2.5mg/L and 1/2MS+IBA1.5mg/L +NAA0.05mg/L were both fit for the culture of rooting.（3）Raising the rate of C/N was advantageous for the culture of rooting.（4）The relative lower concentration of IBA or NAA were advantageous for the culture of rooting，and the effect of IBA was superior to NAA.2 The micropropagation technique of test-tube seedlings（1）The source of carbohydrateAll the test-tube seedlings could grow normally on the mediums which regarded sucrose（chemical purity and analytical purity），glucose or soft white sugar as the source of carbohydrate，fructose was not fit for growth，soluble starch could not be served as the source of carbohydrate；The test tube seedlings could become bulblets on the mediums which regarded sucrose（chemical purity and analytical purity），glucose ，fructose or soft white sugar as the source of carbohydrate，and sucrose of analytical purity was the best one，which the No. and the fresh weight of bulblets came up to 39.67 and 64.93mg respectively，The second one was sucrose of chemical purity，soft white sugar was in the middle，glucose and fructose were the poorest，soft white sugar was a good source of carbohydrate，for growth and bulblet formation，it could become the substitute of sucrose.（2）The concentration of sucroseThe higher concentration of sucrose could inhibit the growth of test-tube seedlings，the seedlings were difficult to form leaves and roots，and were difficult to absorb water，too；Within a certain range，raising the concentration of sucrose could promote bulblets，and the bulblets could form earlier when raising the concentration.But the excessive high concentration（150g/L）could weaken the effect；120g/Lwas the optimal concentration for <WP=11>bulblets，the mean No. and the rate of bulblets came up to 31.33 and 90.00% respectively.（3）PhytohormonesIt was advantageous for bulblet formation when BA and （or）NAA were lower or without phytohormones；NAA1.0mg/L was the best one for the culture of bulblets，the diameter and th fresh weight of bulblets came up to 4.02mm and 115.73mg respectively.（4）PP333Within a certain range，raising concentration was advantageous for bulblet formation，but the excessive high concentration（10.0mg/L）could weaken the effect；The medium of 5.0mg/L PP333 was the optimal one，the mean No.，rate，diameter，fresh weight and index of bulblets came up to 41.00，92.86%，11.55mm，709.06mg and 1.10；PP333 had powerful effect of dumpiness and inhibition， within a certain range，the effect became stronger when raising concentration，the index<1 could be found，and the bulblets became flat round from ellipse gradually ，PP333 could postpone growth，the process of bulblet formation was over 150d；The bulblets on PP333 were light green，and the purplish red peels were relative fewer.（5）S-3307Within a certain range，raising concentration was advantageous for bulblet formation；1.0mg/L S-3307 was the optimal one，the mean No.，rate，diameter，and index of bulblets came up to 45.67，93.34%，8.52mm and 1.21；S-3307 had powerful effect of dumpniss，within a certain range，the effect became stronger when raising concentration，the index<1 could be found，and the bulblets became round from ellipse gradually；S-3307 could postpone growth，the process of formation for bulblets was over 150～210d；The bulblets on更多还原