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

研究通过引入编码对氰化物不敏感的呼吸酶（替代氧化酶）的基因来生产耐寒酶。

• 批准号: 10556001
• 负责人: NAKAZONO Mikio
• 金额: $ 6.66万
• 依托单位: THE UNIVERSITY OF TOKYO
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-10556001/
• 关键词: rice; the cyanide-insensitive respiratory pathway; alternative oxidase; cold-tolerance; altenative oxidase; 遺伝子導入

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.

将编码水稻AOX1a或A0X1b的基因片段通过农杆菌介导法导入水稻(cv.Sasanishiki)。获得了17株AOX1a转基因植株和30株A0X1b转基因植株。其中，我们利用AOX特异性抗体通过Western blotting检测了这些转基因植株中AOX蛋白的含量，最终筛选出三株AOX1a高产或AOX1b高产植株。2000年，我们获得了三种AOX高产植物中的两种的种子。由于我们还没有研究AOX1a或AOX1b高产植株是否具有水稻的耐冷性，我们计划在2001年夏季研究这种可能性。我们通过Northern杂交证明了水稻AOX1a和AOX1b基因的转录在低温、高盐和干旱胁迫下被诱导。但外源ABA不能诱导AOX1a和AOX1b的转录。

项目成果

Li, Y. et al.: "Molecular and cellular characterizations of a cDNA clone encoding a novel isozyme of aldehyde dehydrogenase from rice"Gene. 249. 67-74 (2000)

Li, Y. 等人：“编码水稻醛脱氢酶新型同工酶的 cDNA 克隆的分子和细胞特征”基因。

Ohtsu et al.: "Characterization of a cDNA encoding a novel subunit for cytochrome c oxidase (COX6b) from rice"Breeding Science. 49. 211-215 (1999)

Ohtsu 等人：“编码水稻细胞色素 c 氧化酶 (COX6b) 新型亚基的 cDNA 的表征”《育种科学》。

Nakazono,M. et al.: "Characterization of two cDNA clones encoding rice F_1F_0-ATPase inhibitor proteins, both of which are targeted to mitochondria."Planta. 210. 188-194 (2000)

中园，M.

Hamanaka et al.: "Identification of cDNA encoding cytochrome c oxidase subunit 5c (COX5c) from rice : expression with nuclear-encoded and mitochondrial-encoded COX genes"Genes & Genetic Systems. 74. 71-75 (1999)

Hamanaka 等人：“从水稻中鉴定编码细胞色素 c 氧化酶亚基 5c (COX5c) 的 cDNA：用核编码和线粒体编码的 COX 基因表达”基因

Ohtsu, K. et al.: "Characterization and expression of the genes for cytochrome c oxidase subunit VIb (COX6b) from rice and Arabidopsis thaliana"Gene. 264. 233-239 (2001)

Ohtsu, K. 等人：“来自水稻和拟南芥的细胞色素 c 氧化酶亚基 VIb (COX6b) 基因的表征和表达”基因。