Research for production of the cold-tolerant by introducing the genes encoding the cyanide-insensitive repiratory enzyme, alternative oxidase.

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The cDNA fragment encoding rice AOX1a or A0X1b, which is the terminal oxidase of the cyanide-insensitive respiratory pathway in mitochondria, was in-frame connected to a Ubiquitin gene promoter and then introduced into rice (cv.Sasanishiki) by the agrobacterium-mediated transformation method. Seventeen independent AOX1a-transgenic plants and thirty independent A0X1b-transgenic plants were obtained. Among them, we measured amounts of AOX proteins in these transgenic plants by western blotting using the AOX-specific antibody and finally selected three AOX1a-overproducing or AOX1b-overproducing plants. In 2000, we obtained seeds of two of the three AOX-overproducing plants. Since we have not yet examined whether the AOX1a- or AOX1b-overproducing plants confer on cold-tolerance of rice, we are planning to examine the possibility in summer of 2001.We showed that transcriptions of the rice AOX1a and AOX1b genes were induced under lower temperature, high salt and drought stresses by Northern hybridization. However, the transcriptions of AOX1a and AOX1b were not induced by exogenous ABA.