Development of Hyperspectral Imaging System to Detect Gene Recombinant Callus

本研究旨在发展一套高光谱影像系统，以侦测基因重组植物细胞。由于耐药性基因有可能对生态系统产生负面影响，因此本研究计划使用表达硝酸还原酶（NR）的基因作为标记基因。如果将NR基因导入具有转基因的细胞中，重组基因将显示出比野生型更高的降低硝酸盐浓度的潜力。因此，建立了非破坏性测定愈伤组织中硝酸盐浓度的方法来检测基因重组愈伤组织。在建立的数学模型中，从近红外光谱估算愈伤组织硝酸盐浓度的准确性方面，测定值与估算值之间的相关系数为0。7363.虽然我们尝试用粒子枪法将NR基因导入菠菜细胞，但由于重组条件不合适，不能产生基因重组愈伤组织。在此基础上，对硝酸盐含量较高的愈伤组织（961。0 mg/L）。0 mg/L）的近红外光谱进行分析。用该数学模型预测硝酸盐浓度的准确率为85%，用该模型预测硝酸盐浓度的准确率为87。4%来自PLS-DA

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