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

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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.