Modeling of Pigment Glycoside Production and Its Regulation by Genetic Transformation in Plant Cell Culture.

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The purpose of the study is to construct a novel method to determine the distribution of pigment accumulation among each cell in plant cell culture. As the results of the quantification of this distribution, a model describing pigment accumulation and its variation through culture period was made for the selection of cell lines and the genetic transformation in pigment production pathway.1) Quantification of an anthocyanin production profile in strawberry cell culture :A new method for the quantitative determination of pigment accumulation profiles in each cell using image analysis was constructed and verified as a reproducible and practical method.2) Genetic transformation of anthocyanin formation pathway for the enhancement of the pigment productivity in strawberry cultured cells :It was verified that the addition of phenylalanine as a precursor in the culture medium enhanced the anthocyanin productivity in strawberry cells. As the uptake rate of the precursor into the cells suggests that one of the rate-limiting enzyme in an anthocyanin production pathway is a CHS,the genetic transformation by the introduction of a CHS gene into the strawberry cells was tried to enhance the productivity. For the genetic analysis of a mutant in anthocyanin-producing cell lines, a continuous culture method was applied for the stabilization of gene expression in anthocyanin production pathway. Cloning of specific gene expressed in the mutant was investigated using a proposed method with a continuous culture and cDNA subtraction.