ISOLATION OF NUTRIENT MUTANTS OF PLANT BY T-DNA TAGGING

在这个项目下进行了三项主要实验。1)利用ICP-MS和ICP-AES分析拟南芥叶片微量营养素浓度。添加conc后，每株植物的叶片（干重）被消化约10微克。硝酸。10 ml消化后稀释后的样品用ICP-MS分析B、Mn、Al、Cu、Zn和Mo，用ICP-AES分析Mn、Al、P、Mg、Fe、K和Ca。该方法可在一天内分析200多个样品，用于检测营养植物突变体。2) T-DNA标记线的构建。以pVICE标记载体为载体，利用农杆菌对野生型拟南芥进行转化。采用真空渗透法进行转化。3)植物基因组T-DNA检测。为了了解pVICE的T-DNA区域是否随机整合到植物基因组中，我们利用从个体转化中提取的植物DNA进行了PCR方法和southern分析。将T-DNA区域随机插入到植物基因组中，每个转化基因组中有1-3个T-DNA。

Three major experiments were conducted under this projects.1) Analysis of micronutritent concentration in the leaf of Arabidopsis thaliana by using ICP-MS and ICP-AES.About 10 micro grams of leaf (dry weight) from each plant was digested after adding of conc. nitric acid. Ten ml of digested and diluted sample was analyzed by ICP-MS for B,Mn, Al, Cu, Zn and Mo, and by ICP-AES for Mn, Al, P,Mg, Fe, K,and Ca. More than 200 individual samples can be analyzed by this method in a day for detection of nutritional planr mutants.2) Construction of T-DNA tagging line.Wild type Arabidopsis thaliana was transformed by Agrobacterium having a tagging vector of pVICE.Vacuume infiltration method was applied for transformation.3) Detection of T-DNA in plant genome.To know random integration of T-DNA region of pVICE into plant genome, PCR method and southern analysis were conducted by using plant DNA extracted from individual transformant. T-DNA region was inserted randomly into plant genome and each transformant have 1-3 T-DNA in the genome.