Direct introduction of foreign gene in higher plants

高等植物中直接导入外源基因

• 批准号: 62560005
• 负责人: HIRATA Yutaka
• 金额: $ 1.54万
• 依托单位: Tokyo University of Agriculture and Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1987
• 资助国家: 日本
• 起止时间: 1987 至 1989
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-62560005/
• 关键词: TRANSFORMATION; GENE INTRODUCTION; DIRECT TRANSFORMATION; GRAFT VARIATION; KANAMYCIN RESISTANT; NOPALINE SYNTHASE; ノパリン合成能; β-glucuronidase; DNA; サザンハイブリダイゼ-ション; パリン合成酵素; ノバリン合成酵素

In order to introduce conventionally foreign gene to higher plants, I tried graft method using transgenic tobacco plants which contained kanasycin resistant and nopaline-synthase genes as stock. Non-transgenic normal tobacco plants were grafted on the transgenic tobacco ones. 6 km resistant plants in more than 33000 (0.08%) from the selfed seeds of scion capsules were selected on kanamycin medium (100ug/ml). Four graft transformants had one km gene in the respective one of two homologous chromesomes, one had km genes in homozygous state and the other one had more than 2 km genes (probably 5) in independent state on its chromosomes from the results of test cross experiment. Southern blot analysis of the graft transformants and the progenies show the existence of the maker km and Nos genes and complicated band patterns.It is concluded that the graft transformation is hopeful as a conventional method of gene introduction in higher plant. However, it is further necessary to make clear the mechanism and increase the efficiency.

为了将常规外源基因导入到高等植物中，本研究以转卡那霉素抗性基因和胭脂碱合成酶基因的烟草植株为砧木，尝试了嫁接法。将非转基因正常烟草植株嫁接到转基因烟草植株上。在卡那霉素（100 μ g/ml）培养基上，从接穗蒴果自交种子中筛选出6 km抗性植株33000余株（0.08%）。测交试验结果表明，4个嫁接转化体的两个同源染色体上各有1个km基因，其中1个含有纯合的km基因，另1个含有2个以上（可能5个）独立的km基因。Southern杂交分析表明，嫁接转化子和后代中存在标记km和Nos基因，且带型复杂，表明嫁接转化有望成为高等植物基因导入的常规方法。但是，还需要进一步明确机制，提高效率。

项目成果

平田豊、佐々木英徳、柳下登: "タバコにおける接木による形質転換III接木後代におけるkm抵抗性の遺伝行動" 育種学雑誌. 39巻(別冊2). 356-357 (1989)

Yutaka Hirata、Hidenori Sasaki 和 Noboru Yanagishita：“烟草嫁接转化 III。嫁接后代中 km 抗性的遗传行为”《育种杂志》第 39 卷（特刊 356-357）（1989 年）。

Y.Hirata,H.Sassa and N.Yagishita: "Graft transformation in tobacco(Nicotiana taliacum)" Inoc.of the 6th International longr.of SABRAO. 531-534 (1989)

Y.Hirata、H.Sassa 和 N.Yagishita：“烟草（Nicotiana taliacum）的嫁接转化”Inoc.of the 6th International longr.of SABRAO。

柳下登, 平田豊, 山下恵子, 水上元, 大橋裕: 育腫学雑誌. 32. 236-237 (1982)

Noboru Yanagishita、Yutaka Hirata、Keiko Yamashita、Hajime Mizukami、Yutaka Ohashi：《生驹杂志》32. 236-237 (1982)。

Y. Hirata: "Graft transformation" Kagaku to Seibutsu, 1990, 28, 118-119.

Y. Hirata：“移植转化” Kagaku to Seibutsu，1990，28，118-119。

平田豊,佐々英徳,柳下登: "タバコにおける接木による形質転換II.接木後代におけるKm抵抗性の遺伝行動" 育種学雑誌. 39(別2). 356-357 (1989)

Yutaka Hirata、Hidenori Sassa、Noboru Yanagishita：“烟草嫁接转化 II。嫁接后代中 Km 抗性的遗传行为”《育种科学》杂志 39（第 2 部分）（1989 年）。