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Study on Male Sterility of Plants by Direct Transfer of Mitochondria and mitochondrial DNA

线粒体和线粒体DNA直接转移研究植物雄性不育

基本信息

批准号：
01560120
负责人：
MORIKAWA Hiromichi
金额：
$ 1.15万
依托单位：
Hiroshima University (1990)Kyoto University (1989)
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1989
资助国家：
日本
起止时间：
1989 至 1990
项目状态：
已结题

项目摘要

Particle bombardment has been shown to be a powerful method for the transformation of organelles by the previous authors (e. g., Svab et al. PNAS 87 : , 8526-8530, 1990). In this study we developed our own particle gun device and attempted to transform mitochondria of higher plants. We have not yet succeeded in transformation of mitodondria. However, we obtained some fundamental results that will be useful for studies on organeLle transformation in future.1. We have developed particle gun devices that are driven by nitrogen gas pressure and compressed air pressure. Nitrogen-gas-pressure-driven device had a maximum accelerating pressure (i. e. pressure that was used to accelerate plastic projectiles) of 25 kg/cm^2, while pneumatic particle gun device had a maximum accelerating pressure of more than 220 kg/cm^2. With the pneumatic particle gun, the maximum velocity of projectile was more than 440 m/s.2. Using these devices, we have shown that plasmid DNA can successfully be introduced and the introduced foreign genes (e. g. GUSgene) can successfully be expressed in intact tissues as well ascultured cells of various plants such as tobacco, soybean and Arabidopsis (dicots) and rice and wheat (monocots).3. We succeeded in stable transformation of cultured cells of tobacco with NPTII gene by use of these two particle gun devices. We also delivered the NPTII gene into root cells of Arabidopsis and selected kanamycin-resistant shoots. The selected shoots successfully developed into complete plants (more than 20 such plants have so far been obtained) in the presence of kanamycin and set seeds.

粒子轰击已被证明是一个强大的方法转化的细胞器由以前的作者（e。例如，在一个实施例中，Svab等人PNAS 87：，8526-8530，1990）。在本研究中，我们开发了自己的粒子枪装置，并尝试转化高等植物线粒体。我们还没有成功地改造线粒体。但是，我们也得到了一些基础性的结果，这些结果对今后的细胞器转化研究具有一定的参考价值.我们已经开发出由氮气压力和压缩空气压力驱动的粒子枪装置。氮气压力驱动装置有一个最大加速压力（即：e.用于加速塑料射弹的最大压力为25 kg/cm^2，而气动粒子枪装置的最大加速压力超过220 kg/cm^2。采用气动粒子炮，弹丸最大速度大于440 m/s.使用这些装置，我们已经表明质粒DNA可以成功地被引入，并且引入的外源基因（e. G. GUS基因）在烟草、大豆、拟南芥（双子叶植物）和水稻、小麦（单子叶植物）的完整组织和培养细胞中均能成功表达.利用这两种粒子枪装置，我们成功地将NPTII基因稳定转化烟草培养细胞。我们还将NPTII基因导入拟南芥的根细胞中，并选择了卡那霉素抗性芽。在卡那霉素存在下，所选择的芽成功地发育成完整的植物（迄今为止已经获得了20多株这样的植物）并结出种子。