TRANSFORMATION SYSTEM INFRUIT TREES USING T-DNA REGION OF AGROBACTERIUM FOR REGENERATION EFFICIENCY

为开发果树遗传转化方法，以日本梨为材料，对农杆菌二元体系进行了研究。在日本梨的几个栽培品种和野生种中，发现野生种中有一个品系具有快速繁殖能力，适合以茎叶为转化材料。Gerlite作为组织培养的支撑培养基，配合微滤池调节培养瓶的相对湿度是有效的。Tidiazuron也可以有效地代替苄基腺嘌呤，并且也需要微过滤器附着在培养瓶的盖子上。种子胚芽是芽部再生能力强的材料。以农杆菌EHA101和pIG121为二元转化体系进行转化。观察Gus表达以证实转化。在转化早期，茎端切除后进行4天的预培养是有效的。共培养天数对子叶侵染有影响。叶片解剖部位细菌侵染，叶片表面气孔细胞丰富的地方出现斑纹。细菌浓度

To develop the method of genetic transformation in fruit trees, Agrobacterim binary system was studied using the Japanese pear. Among the several cultivers and the wild species of the Japanese pear, one of the strain of the wild species was found to have a rapid multiplification suitable for subjecting the shoot and leave for transformation materials. Gerlite as a supot medium for tissue culture was effective when it was used with microfilter in adjusting the relative humidy of the culture bottle. Tidiazuron was also effective in place of benzyladenine and also required the microfilter attached to the cap of the culture bottle. Seed coleoptiles were sujected for highly shoot regreneration materials. Transformation was carrid with Agrobacterim EHA101 strain with pIG121 as a binary transformation system. Gus expression was observed to confirm the transformation. In eary stage of the transfomation, pre-culture of the shoots for four days after discetion of the stem end was effective. Co-cultivation days were infuence for the infection of cotyledon. Leaf disk was infected with bacteria at the disceted part and the gus spots were observed on the disk surface where the stomata cells were aboundant. Concentration of the bacteria