Analysis and proposal of new aluminum resistanece strategy by developing of model technique using artificial cell membrane

目标已基本完成，最终可以提出新的抗铝策略。我承认通过我的研究建议的出色判断。本文的研究重点如下。（1）原生质体分离新技术的建立将根尖分离的原生质体粘附在多聚赖氨酸包被的玻璃板上，用缓冲液洗涤。发现粘附的残余膜几乎不含其他细胞器膜。最后，我可以开发一个新的简单的技术，分离PM相比，传统的液体两相分配技术。（2）PM脂质包被尼龙胶囊渗透性实验体系的建立合成了尼龙-2，8粒胶囊，并在其膜上的微孔上包被了脂质双层。该实验系统上级传统的脂质体系统。根系释放有机酸阴离子不明显， 关于我们 抗铝小黑麦的质膜脂质在铝介质中的渗透性明显低于铝敏感小黑麦，而抗铝小黑麦的质膜脂质富含游离甾醇和葡萄糖脑苷脂。（3）根尖原生质体亚甲基蓝（MB）染色强度与植物抗铝性的关系在许多植物种、品种和品系中，原生质体MB染色强度与抗铝性呈显著负相关。这说明抗铝植物根尖的负性较低，质膜较强。（4）豌豆油菜素内酯（BR）突变体和赤霉素（GA）突变体对铝的抗性比较在几个豌豆BR突变体和GA突变体中，Ih突变体对高铝胁迫最为敏感。这种最高的敏感性与PM的最大渗透性相关，但不与OA从根中释放。这可能与PM脂质层中游离甾醇异常积累有关。（5）信号草的耐高铝性信号草根尖质膜透性较低。新发现的高沉积类黄酮周围的根尖细胞的周边区域，这也可以有助于信号草PM的渗透性最小。少

The objectives have almost been completed, and finally a new aluminum resistance strategy could be proposed. I acknowledge the excellent judgement on the adoption of my research proposal. The points of my research are as follows.(1)Development of new and simple technique for isolation of plasma mebrane(PM)Protoplasts isolated from root-tips were adhered on polylysine coated glass plates, and washed with buffer solution. The adhered residual membranes were found to be almost free from other organelle membranes. Finally, I could develop a new simple technique for isolation of PM as compared with conventional liquid two polymer phase partitioning technique.(2)Establishment of permeability experiment system using synthesized nylon capsule coated with PM lipidNylon-2, 8 capsules were synthesized and the micropores in their membranes were coated with lipid bilayer. This experimental system was superior to the converntional liposome system. Organic acid anion release from roots was not signif … More icantly different between two lines of triticale, but on the other hand, PM lipid which is qualitatively rich in free sterols and glucocerebrosides of Al-resistant triticale line was less permeable than that of Al-sensitive line in Al medium.(3)Association of methylene blue(MB)stainability of protoplasts isolated from root-tips with Al resistanceSignificant negative correlation was recognized between Al resistance and MB stainability of protoplasts among many plant species, cultivars and lines. This suggests that Al resistant plants have their root-tips with lower negativity and stronger PM.(4)Comparison of Al resistances among brassinosteroid(BR) mutants and gibberellin (GA) mutants of peaAmong several BR mutants and GA mutants of pea, Ih mutant was most sensitive to high Al conditions. This highest sensitivity was correlated with the greatest permeability of PM, but not with OA release from roots. This may be associated with the abnormal accumulation of free sterols in PM lipid layer.(5)Hyper Al resistance of signalgrassPermeability of root-tip PM for signalgrass was considerably low. High deposition of flavonoids around the peripheral area of root-tip cells was newly found ; this can also contribute to the least permeability of signalgrass PM. Less