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

为了建立果树遗传转化的方法，以日本梨为试材，研究了农杆菌二进制转化系统。在日本梨的几个栽培者和野生种中，发现其中一个野生种的品系具有快速繁殖能力，适合作为转化材料的试管苗和叶片。利用微滤器调节培养瓶的相对湿度，是一种有效的组织培养Supot基质。替地卓隆也能有效地替代苄基腺嘌呤，而且还需要在培养瓶盖上安装微过滤器。种子胚芽鞘被用作高再生材料。以农杆菌EHA101菌株为转化载体，以pIG121为二元转化系统进行转化。观察到GUS的表达证实了转化。在转化的早期阶段，茎端切除后预培养4天是有效的。共培养天数对子叶的侵染有影响。叶盘被细菌侵染，气孔细胞丰富的叶盘表面可见GUS斑点。细菌的浓度

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