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

编码水稻AOX1a或A0X1b的cDNA片段是线粒体中氰化不敏感呼吸途径的末端氧化酶，该片段与泛素基因启动子在框架内连接，然后通过农杆菌介导的转化方法导入水稻（cv.Sasanishiki）。获得17株独立的aox1a转基因植株和30株独立的a0x1b转基因植株。其中，我们利用aox1特异性抗体，通过western blotting测定了这些转基因植株中AOX蛋白的含量，最终选择了3株aox1a过量或aox1b过量的植株。2000年，我们获得了三种aox过量生产植物中的两种的种子。由于我们还没有研究过量生产AOX1a-或aox1b -的植物是否会对水稻的耐寒性产生影响，我们计划在2001年夏季研究这种可能性。通过北方杂交，我们发现在低温、高盐和干旱胁迫下，水稻AOX1a和AOX1b基因可以被诱导转录。然而，AOX1a和AOX1b的转录不受外源ABA的诱导。

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.