【作者】 刘磊；

【导师】 白志川； 刘会超；

【作者基本信息】 西南大学 ， 植物学， 2009， 硕士

【摘要】 本研究以牡丹成熟胚、花药、鳞芽、未成熟胚为外植体,建立了牡丹成熟胚经愈伤组织途径分化不定芽的完整再生体系;通过对牡丹花药进行预处理,成功诱导出了花药愈伤组织;经过改进消毒方法,解决了牡丹鳞芽污染严重的问题;研究了环境因子、植物生长调节剂在牡丹鳞芽生根过程中的作用,建立了鳞芽生根体系;探讨了培养因子在牡丹’洛阳红’未成熟胚成苗过程中的作用,筛选出适宜于牡丹组培苗移栽驯化的基质类型和配比。主要研究结果如下:1.牡丹’凤丹白’成熟胚不定芽诱导和生根的研究结果表明:在打破种子休眠方面,以500mg·L^-1GA₃浸泡48h效果最好:1/2MS+Ca²⁺为’凤丹白’胚培养的最适基本培养基;植物生长调节剂组合为2,4-D40mg·L^-1+6-BA0.1mg·L^-1时,愈伤组织的诱导效果最好;6-BA诱导愈伤组织分化不定芽的最佳浓度为1.0mg·L^-1;诱导不定芽生根的最佳培养基为:1/2MS+IBA1.5mg·L^-1+NAA0.5mg·L^-1。2.牡丹’乌龙捧盛’花药培养的研究结果表明:未做任何预处理的花药,其愈伤组织平均诱导率明显高于经过低温或高温预处理的花药（P<0.05）;最佳的诱导培养基为:MS+2,4-D2.0mg·L^-1+6.BA1.0mg·L^-1,愈伤组织的平均诱导率最高,达到36.87%。3.牡丹’乌龙捧盛’鳞芽组织培养的研究结果表明:采用0.2%（w/v）HgCl₂进行牡丹鳞芽消毒处理的适宜时间为6-8min;鳞芽初代培养过程中,先暗培养7d,而后转入正常光照培养,可明显减轻鳞芽褐化;诱导根原基形成的最适培养基为:1/2MS+NAA2.0mg·L^-1+IAA2.0mg·L^-1+6-BA0.1mg·L^-1或1/2MS+NAA1.0mg·L^-1+IBA1.0mg·L^-1+AC0.2g·L^-1。4.牡丹’洛阳红’未成熟胚培养和植株再生的研究结果表明:诱导牡丹’洛阳红’未成熟胚成苗的最佳培养基为:1/2MS+6-BA0.1mg·L^-1+IBA0.8mg·L^-1+LH100mg·L^-1。组培苗移栽驯化的适宜基质为:腐殖土:洛阳园土=1:1。更多还原

【Abstract】 Peony mature embryo , anther, sprout, immature embryo are taken as the explants in this paper. Complete multiple shoots regeneration system was established by the way of callus induction. Callus was obtained successfully by the way of anther ,and the anther was pretreated . According to the selection of disinfection ,the effective method against contamination was obtained.The induced rooting of sprout under some enviroment factors and plant growth regulators were researched,and the induced rooting of sprout complete system was established. Some culture factors under seedling formation of immature embryo ’Luo Yang Hong’ were discussed,the appropriate medium type and mixture for tissue culture seedling domestication were selected.The main results were as follows:1.500 mg?L^-1GA₃ soaking treatment 48h was the best effective treatment in the aspect of breaking dormancy. 1/2MS+Ca²⁺ was the most appropriate basic medium in the culture of peony embryo. When the medium contain 2,4-D 4.0 mg?L^-1+6-BA0.1mg.L^-1callus was introduced well. The most optimum 6-BA concentration for introduction of multiple shoot from callus was 1.0 mg·L^-1. The most optimum medium for plantlet regeneration was 1/2MS+IBA 1.5 mg·L^-1+NAA0.5mg·L^-1.2. The average frequencies of callus induction in the control group was obviously higher than cold pretreatment group and high temperature pretreatment group （P<0.05）.The best medium of multiplication was MS+2,4-D 2.0 mg?L^-1+6-BA1.0 mg?L^-1, which average frequencies of callus induction reached 36.87%.3. The appropriate disinfecting time for sprout is 6-8min if use 0.2%（w/v）HgCl₂.During the course of initial culture,first, sprout was cultured under dark for 7d,and then sprout was cultured under normal light condition,and browning was relieved obviously by the. way. The most optimum medium for root primordium was:1/2MS+NAA 2.0 mg·L^-1+IAA2.0mg?L^-1+6-BA 0.1mg·L^-1orl/2MS+NAA1.0mg·L^-1+IBA1.0mg?L^-1+AC0.2g?L^-1.4. The best medium for seedling formation of peony ’Luo Yang Hong’ was: 1/2 MS+6-BA 0.1mg·L^-1+IBA 0.8 mg·L^-1+LH 100 mg·L^-1. Appropriate matrix for domestication was humus soil:’Luo Yang’ soil=1:1.更多还原