Development of Hyperspectral Imaging System to Detect Gene Recombinant Callus

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The purpose of this research is to develop a hyperspectral imaging system to detect gene recombinant plant cells. Since drug tolerance genes have the potential to negatively affect ecosystems, this research plans to use genes that express the nitrate reductase(NR) as marker genes. If the NR genes are introduced into a cell with a transgene, the recombined genes would show higher potential to reduce nitrate concentration than that of a wild type. Therefore, non-destructive methods for measurements of nitrate concentration in callus were developed to detect the gene recombinant calluses. As for accuracy of developed mathematical model to estimate the nitrate concentration of callus from the near-infrared spectrum, correlation coefficient between measured and estimated nitrate concentration became 0. 7363. Though we tried to introduce the NR gene to spinach cells by using the particle gun method, gene recombinant calluses could not be produced because of inappropriate conditions for the recombination. Then possibilities to discriminate calluses containing high nitrate concentration(more than 961. 0 mg/L) from low concentration calluses(less than 961. 0 mg/L) by processing the near-infrared spectra were examined. Accuracy rate became 85% that was obtained from the mathematical model to predict the nitrate concentration and 87. 4% that was derived from PLS-DA