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利用水稻耐盐品种兰胜构建了一个高质量的cDNA文库以鉴定水稻盐胁迫应答基因,从cDNA文库中提取约15000个质粒,并用Biomek 2000高密度点阵系统或手工操作,将这些质粒点于尼龙膜上.通过这种方法鉴定了30个盐应答基因,对其中的12个基因通过Northern杂交进行表达分析,确证了cDNA微阵列的杂交结果.30个基因中,18个基因受盐诱导,另外12个基因受盐抑制,其中27个在GenBank数据库中有同源序列.根据功能,这些基因大致可以分为5类:光合作用相关基因、物质运输相关基因、代谢相关基因、耐逆相关基因以及其他未分类的基因.研究结果表明,盐胁迫影响了植物生长发育的多个方面,其中有些基因可能在植物体的耐盐过程中具有重要作用.
A high quality cDNA library was constructed using a salt tolerant variety, Lansheng, to identify rice salt stress response genes. Approximately 15,000 plasmids were extracted from the cDNA library and used Biomek 2000 high-density dot matrix system or hand-operated to spot these plasmids on Nylon membrane.Through this method, 30 salt-responsive genes were identified and 12 of them were analyzed by Northern blot to confirm the hybridization results of cDNA microarray.Of the 30 genes, 18 genes were induced by salt, The other 12 genes were inhibited by salt, of which 27 had homologous sequences in the GenBank database.According to the function, these genes can be divided into five groups: photosynthesis related genes, transport related genes, metabolism related genes, resistance-related genes And other unclassified genes.The results show that salt stress affects many aspects of plant growth and development and some of these genes may play an important role in the salt tolerance of plants.