【作者】 吕校石；

【导师】 杨建民；

【作者基本信息】 河北农业大学 ， 森林培育， 2004， 硕士

【摘要】 椴树树形美观，叶大荫浓，叶形多变，寿龄长，花芳香馥郁，病虫害少，对烟尘及有害气体抗性强，是优良的园林观赏树种。为了满足椴树对绿化的需要，尽快获得大批量绿化优质用苗，本文利用组织培养技术建立了引进的彩叶椴树金叶椴，皇帝椴和紫椴、蒙椴，糠椴的再生体系。主要从外植体的消毒和选择、基本培养基的应用、外源激素的使用浓度、影响试管苗生根的主要因子以及试管苗的移栽等几个方面进行了探讨，确定了最佳再生途径。主要结果如下： 1 污染控制 以椴树的茎尖作外植体，影响其污染率的主要因素是取材的季节不同，污染率依次是：4～5月份＜6～7月份＜8～9月份＜10月后。因此，外植体应尽早采取抽生的新芽或萌蘖。 椴树的茎尖对升汞敏感，出现腋芽坏死现象。以安替福民、青霉素、链霉素，84消毒液为消毒剂进行了一系列的消毒实验，结果表明：84消毒液对外植体的消毒效果优于安替福民。以10％的84消毒液处理种子30min或15％的84消毒液处理种子20min或以15％的84消毒液处理茎尖30min，效果最佳，污染率均为0。用320万单位／L的青霉素+160万单位／L的链霉素浸泡35min，或用480万单位／L的青霉素+160万单位／L的链霉素浸泡水培苗30min控制细菌污染，效果最佳。 2 胚的培养 以半成熟的种子为组培材料，不仅利于操作，而且有利于胚的生长。在保定市区，应在8月5日左右；白石山山区，则在9月1日左右采集紫椴种子。较适宜的培养基为MS+0.5mg／LKT+0.05mg／LNAA+2.0mg／LGA₃。KT的适宜浓度为0.5mg／L，高于或低于这个浓度都影响胚的生长。 3 茎尖的启动 茎尖的质量和取材的时间是启动成败的重要因素。应采取当年生半木质化嫩茎或萌蘖为材料，不仅利于启动，又利于茎尖的生长和分化；在取材的时间上，以4～5月份最佳，6～7月份也较理想。不同的基因型，其启动率不一样。紫椴、蒙椴＞皇帝椴＞金叶椴、糠椴。较适宜椴树启动的培养基为MCM+1.0mg／LBA+0.1mg／LNAA。 4 增殖培养和褐变控制 椴树的增殖的基本方式是以通过种胚获得的无菌苗和茎尖启动获得的腋芽进行继代培养，以获得最高的繁殖系数，通过控制培养基中分裂速度浓度来完成对读数试管苗生长状况的调控。实验表明：在紫椴、蒙椴增殖培养中，BA对茎尖的生长和分化表现稳定，其中以WPM+0.05mg／LBA+2.0mg／LGA₃+0.05mg／LNAA培养基中茎尖的增殖系数较高，苗生长旺盛，茎粗日十大，且成本较低。ZT对椴树茎尖的生长和分化与BA没有明显的差异，且成本较高。在金叶椴增殖培养中，TDZ表现出很强的促进分化和增殖生长能力。其中以WPM+0.05mg／LZT+2.0mg／LGA₃+0.5mg／LNAA培养基中茎尖的增殖系数最高。 引起褐变现象的因子是多方面的，试验过程中，在培养基中加入0.1mg／L活性炭+0.1PVP或1.0g／L硫代硫酸钠+0.1g／L活性炭有效地抑制椴树褐变的发生 5 生根培养和移栽 影响试管苗的生根因素很多，比较各种激素种类和浓度对锻树试管苗的生根影响。实验结果表明:锻树试管苗直接诱导生根效果较差;该用二步生根法效果理想，其中以l/ZMs+15om泌IBA巧omg毛N从诱导3d再转入生根培养基中生根效果最佳:紫锻和糠锻的生根率为100%，檬锻的生率为90.0%，金叶锻的生根率为54.0%，根质量高，粗壮、亮白色、生长迅速，无侧根。 锻树试管苗移栽基质采用营养土:细沙=2:1的效果最佳。移栽的成活率在一定程度上与移栽试管苗的根的活动状态有关，试管苗根的长度和生长状态是影响试管苗成活的重要因素，当茎尖苗根长为1 .0一2.ocm时，其成活率为95.8%，原因这正是根生长最快的时期，因而其成活率最高。更多还原

【Abstract】 The Tilia is fine garden ornamental ,which shape is beauty and the leaf is big .umbrage is great ,the shape of leave is levity and the life is long .at the same time the spiciness of the flower is strong ,the plant diseases and insect pests are litter the fastness is strong to soot and deleterious gas.To fulfill in needing of vivescence and odtain lots of tilia seedling,In.this paper studies on tissue culture of T.platyphyllos ’Aurea’ , Tilia europaea ’Pallida , Tilia mands -hurica Rupr.et Maxim , Tilia amurensis Rupr, Tilia mongolica Maxim. We assessed the effects of genotypes ,the type of explants ,instiation medium,hormones and explants. Physiological stages in course of adventitious bud regeneration. Regeneration. System of birch was estab -lished in vitro.1 Pollution ControlUsing tilia stem as explant of tissue culture .The ratio of pollution is season.4~5 months <6~7months<8~9months<after10,so we pick the new bud and sprout as early as possible.The tilia stem is sensitive to Hgcl the bud of tilia dies after dealing with it. so we stenrilize tilia with NaClO,penicilline,streptomycinand 84 disinfector. The result indicates:using steril aqueous solution of penicillin with 3.2 million and streptomycin with 1.6 million in 30 minutes or using steril aqueous solution of penicillin with 4.8million and streptomycin with 1.6 million in 35 minutes to deal with sprout cultivated in water after dealing with NaClO with 10%.Their polluted ratios are 3%. The effect are very ideal. Using 84 disinfector with 10% deal with seeds in 30 minutes or 15% deal with seeds in 20 minutes .Their polluted ratios are 0% The effect are the best ideal.2 Embryo culturewe should use the half mature seeds of Tilia , wich not only is propitious to maniplation but also is fvour of growth of embryo .we should pick seed adout 5/8 in baoding downtown or about 1/9 in baishi montain. The better medium is MS macronutrients with 0.5mg/LKT+0.05 mg/LNAA+2.0mg/LGA3. the rigt concentration of KT is 0.5mg/LKT,high and less concentr -ation of KT can effect the percentage of adventitious bud indution.3 Startupo of stemThe qulity and season are important fators of startup we should pickthe half lignifacationnew buds and spouts which not only is propitious to maniplation but also is fvour of growth andpolarization of embryo.In season effect of 4-5 month is the best and 6~7 month is better. Ingenotype Tilia amurensis Rupr. And Tilia mongolica Maxim.are superio toTilia europaea’Pallida T.platyphyllos ’Aurea’ and Tilia mandshurica Rupr.et Maxim are badly The bettermedium is MS macronutrients with 1.0mg/LBA+0.05mg/LNAA.4 Culture of multiplication and control of browningThe basal method of multipication of birch clones adventitions bud oy yong twig is culturing offspring with way of overgrow buds.which can get the highest multiplication rate.the concentration of CTK in the medium regulated growth of birch seedlings in vitro. Experiment -ation indicates: the ability of growth and polarization to stem of Tilia amurensis Rupr. And Tilia mongolica is stablization.The effect of WPM+0.05mg/L+2.0mg/L+0.05mg/L is the better,the growth is blooming the stem is gruff,leave are big and the cost is litter.the effection of ZT is the same as BA but it cost high .In the experiment using ZT to deal with T.platyphyllos ’Aurea’ ,the ability of gowth and polarization is very strong. And the effect of culture medium WPM+0.05mg/LZT+2.0mg/LGA3+0.5mg/LNAA is better.There are many factor effect browning In experiment using 0.1 g/L active carbon+0.1mg/L PVP or 0.1g/L active carbon+1 .0g/LNaS2O3 can control browing effective5 Rooting culture And TransplantingThere are many factors that effected Tilia in vitro on rooting Compare all kinds of hormones and the thicknes which effected Tilia in vitro on rooting .The results showed Tilpa in vitro is not suit for rooting directly It is better for two steps of rooting.including 1/2MS+150mg/LIBA+50mg/LNAA medium can best induce adveneration root differentiation. Which root is good ,strong white,grown quite ,no更多还原