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

最后提出了一种新的抗铝策略。我感谢你们对我的研究计划的采纳做出了卓越的判断。我的研究要点如下。(1)建立了一种简便的分离根尖质膜（PM）原生质体的新技术，将分离得到的PM原生质体粘附在聚赖氨酸镀膜玻璃板上，并用缓冲液清洗。发现粘附的残余膜几乎不受其他细胞器膜的影响。最后，与传统的液体双聚合物相分配技术相比，我可以开发一种新的简单的分离PM的技术。(2)采用PM脂包被合成尼龙胶囊建立渗透性实验体系，合成尼龙- 2,8个胶囊，并在其膜上包被脂质双分子层的微孔。该实验体系优于传统的脂质体体系。根中有机酸阴离子释放量在两系小黑麦间差异较大，但抗铝小黑麦品系的PM脂质在铝介质中的渗透性比铝敏感品系低，PM脂质富含游离甾醇和糖苷。(3)根尖原生质体抗铝性与亚甲基蓝（MB）染色性的关系。在许多植物种类、品种和品系中，原生质体抗铝性与亚甲基蓝染色性呈显著负相关。这说明抗铝植物的根尖负性较低，PM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豌豆BR和GA突变体中，Ih突变体对高铝条件最敏感。这种最高敏感性与PM的最大渗透性相关，但与根部OA释放无关。这可能与游离固醇在PM脂质层的异常积聚有关。(5)信号草的抗超铝能力，根尖PM对信号草的渗透性较低。新发现根尖细胞周围黄酮类化合物大量沉积；这也可能导致信号草PM的渗透率最低。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