A SEARCH FOR THE KEY MATERIAL INVOLVED IN SOYBEAN SUPERNODULATION.

1)建立了一种高效收集大豆Williams超结突变体NOD 1-3和Williams82韧皮部汁液溶质的方法。通过EDTA预处理或同时处理大豆离体芽，收集了韧皮部汁液溶质，并明确了有效收集的条件。研究了从韧皮部汁液溶质收集液中去除EDTA的方法，在冰冷和酸性条件下，EDTA大部分以沉淀物的形式被去除。2)建立评价韧皮部液溶质调控根瘤形成活性的生物测定体系。扦插NOD1-3和William82三联叶，获得植株。不同品种的植株分别表现出超结瘤和正常结瘤的性状。通过真空入渗将蔗糖或尿素单独引入植株叶片，可以明显改变根系根瘤和根瘤样块的数量。d.在引入尿素的情况下，重氮示踪技术也证实了尿素产生的氮被大量输送到根系。因此，植株-真空渗透体系可用于评价韧皮部液溶质对根瘤形成的调节作用。3) NOD1-3和William82的韧皮部汁液溶质组成及控制根瘤形成的关键物质的分离。在游离氨基酸组成上，NOD1-3与Williams82无论结瘤与否均存在显著差异。在糖的组成上，两者无显著差异。CDS柱高效液相色谱分析显示NOD1-3与Williams82之间存在一些含量差异较大的不明物质。然而，在生物测定苗的韧皮部液溶质导入检测中，未能检测到控制根瘤形成的关键物质。少

1) Establishment of an efficient method for the collecting phloem sap solutes from soybean NOD 1-3(a supernodulating mutant of Williams) and Williams82.By pre- or simultaneous treatment of excised soybean shoots with EDTA, phloem sap solutes were collected, and the conditions for the efficient collection were clarified. A method for removing EDTA from the collected liquid of phloem sap solutes was examined, and most of EDTA was removed as precipitate by standing the liquid under ice-cooled and acidic condition.2) The establishment of a bioassay system for evaluating the phloem sap solutes in the activity of regulating root nodule formation.Plantlets was produced by cuttage of NOD1-3 and William82 triplet leaves. The plantlets respectively showed supernodulating or normal nodulating trait corresponding to catage source. By the individual introduction of sucrose or urea into the plantlet leaf through vacuum infiltration, the number of root nodule and root nodule anlage was clearly change … More d. In case of the urea introduction, it was also confirmed by heavy nitrogen tracer technique that the nitrogen derived from urea was abundantly transported to the roots. Hence, it was suggested that the plantlet-vacuum infiltration system was applicable for evaluating the phloem sap solutes in the activity of regulating root nodule formation.3) Composition of phloem sap solutes collected from NOD1-3 and William82, and isolation of key material which controls root nodule formation.On the free amino acid composition, some remarkable differences were detected between NOD1-3 and Williams82 regardless of root nodulation or non-nodulation. On the saccharide composition, there was no remarkable difference between them. An CDS column HPLC revealed that there were some unidentified materials which were much different in content between NOD1-3 and Williams82. However, key material, which controls the root nodule formation, could not be detected in the examination of phloem sap solute introduction into bioassay plantlets. Less