【作者】 贺窑青；

【导师】 范国强； 黎明；

【作者基本信息】 河南农业大学 ， 森林培育， 2003， 硕士

【摘要】 悬铃木是我国园林绿化的主要树种之一，具有较强的净化空气和遮阴能力，观赏性较强，木材可供建筑和家具等用，是集生态、经济和社会效益为一体的品质优良的树种。但随着树龄增大，其果毛污染成为社会环境的一大公害，为通过植物组织培养的方法从理论上研究悬铃木的不育机理，同时为不育转基因悬铃木的培育建立一个良好的基因转化受体系统，本文以悬铃木的叶片为外植体，通过正交试验和单因子试验，借助于细胞组织学观察，研究了不同基本培养基、植物激素不同浓度组合、不同温度处理、叶位和培养时间对诱导叶片形态发生及植株再生的影响，结果表明：叶片诱导植株再生的最佳基本培养基为WPM，非胚性愈伤组织诱导的最佳培养基为WPM+0.6IBA+8.06-BA、适宜培养20天，胚性愈伤组织诱导的最佳培养基为WPM+0.1IBA+5.06-BA、适宜培养25天，非胚性愈伤组织诱导不定芽的最佳培养基为WPM+0.1IBA+5.06-BA、适宜培养50天，胚性愈伤组织体细胞胚胎发育的最佳培养基为WPM+0.56-BA、适宜培养45天，叶片不定芽诱导的最佳培养基为WPM+0.1IBA+3.06-BA、适宜培养50天，最佳根诱导培养为1／2Ms0.1—0.5IBA、最佳胚状体苗发育的培养基为WPM；接种的叶片组织或愈伤组织在经10℃低温预处理3天后，置于25℃温度条件中培养，可不同程度的提高诱导率；叶片植株再生所用外植体以无菌苗的第七片真叶完全展开后从其基部往上数第五片叶不定芽诱导率最高；在培养过程中，生长素IBA有利于愈伤组织的诱导，细胞分裂素6-BA有利于不定芽的诱导，ZT对愈伤组织及不定芽的诱导有抑制作用；在开始对叶片愈伤组织的诱导中需要较高浓度的生长素和细胞分裂素，在诱导不定芽和体细胞胚时需降低它们的浓度，而在体细胞胚胎发育过程中不需要生长素，仅附加细胞分裂素；米黄色、微紧的愈伤组织是器官发生的主要愈伤组织，褐、紫红色、微松的愈伤组织是体细胞胚胎发生的主要愈伤组织，这些结果为悬铃木采用不同的方式培育新品种提供了基础。更多还原

【Abstract】 Platanus orientalis L. is one of main gardens greening tree species in our country. It has the stronger ability of cleaning air and stronger ornamental characteristic. Its wood may be used as architecture and furniture and so. It is a good quality tree species having ecological, economical and society benefit. But with tree age older, its fruit hair’ contamination has become a big public nuisance. In order to study mechanics of Platanus orientalis L. sterility from theory by plant tissue culture and build a receptor system of good gene inversion for sterility trans-gene Platanus occidentalis L. culture, the paper used Platanus orientalis L. leaves as explants and studied the effects of different basic culture media, different concentration of plant hormone ,different temperature treatment, leaf position and culture time on inducing leaf morphogenesis and plant regeneration by orthogonal layout and signal factor experiment. The results showed: the best basic culture media was WPM.The best culture media of no-embryo callus induction was WPM+0.16IBA+8.06-BA.The best culture media of 20d suitable culture, embryo callus tissue induction was WPM+0.1IBA+5.06-BA.The best culture media of 25d suitable culture, no-embryo callus tissue induction was WPM+0.1IBA+5.06-BA. The best culture media of 50d suitable culture, embryo callus tissue somatic embryogenesis was WPM+0.56-BA. The best culture media of 45d suitable culture, adventitious bud induction was WPM+0.1 IBA+3.06-BA. The best culture hiedia of 50d suitable culture, the best culture media of root induction and embryoid development was 1/2MS0.1~ 0.51BA and WPM separately. 3d after inoculated leaves tissue or callus were pre-treated in 10癈 low temperature, the inducting ratio was improved differently under 25 癈 temperature conditions. The highest ratio of leaf, plant regeneration’ adventitious bud induction is the fifth leaf numbering from basis to top after the seventh true leaf of germ-free seedlings unfolded. During culture, hormone IBA benefited callus induction, CTK-6-BA benefited adventitious bud induction, the effects of ZT on callus and adventitious induction wasn’t significant.At first, IAA and CTK of higher concentration were needed in leaf callus induction. Their concentrations were needed to reduce in inducting adventitious. But IAA was’t needed in somatic embryo44development, only needed to add CTK. Rice-yellow, tight callus was organogenesis main callus. Brown, purble-red, loose callus was the main callus of somatic embryogenesis.These results supplied basis for the culture of Plalanus occidenlalis L. new varietyby different means.更多还原