ISOLATION OF NUTRIENT MUTANTS OF PLANT BY T-DNA TAGGING

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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.