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

1996 Fiscal Year Final Research Report Summary

• Project/Area Number: 07806002
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: 園芸・造園学
• Research Institution: KOBE UNIVERSITY
• Principal Investigator: NAKANISHI Tetsu GRADUATE SCHOOL OF SCIENCE AND TECHNOLOGY, 自然科学研究科, 教授 (80031227)
• Project Period (FY): 1995 – 1996
• Keywords: JAPANESE PEAR / TRANSFORMATION / TISSUE CULTURE / AGROBACTERIUM TUMEFACIENS

Research Abstract

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