Changes in Flavonoid Ingredients in Micro-Tom Tomato Transgenic Plants Introduced with Citrus CuCYP71A1 Gene

Abstract

In this study, we researched the citrus gene cuCYP71A1 developed from the red citrus mutant pigment is introduced into the Micro-Tom tomato seeds using pcambia2300vector. T2 generation seeds were sowed in the greenhouse, genomic DNA was extracted, and the introduction of the target gene was confirmed through genotyping PCR analysis. After confirming the gene introduction, tomato homozygous were prepared, sown in the greenhouse, and harvested by dividing leaves, flowers, and 6 fruit development stages. The color difference was taken for each stage of the fruit using a colorimeter, RNA was extracted, and cDNA synthesized. After cDNA synthesis, RT-PCR analysis confirmed the expression of the target gene. As a result, it was confirmed that the citrus gene cuCYP71A1 introduced into tomatoes was expressed in all stages. After the introduction and expression of the target, the gene was confirmed as described above, the expression levels of the tomato-related genes SLCYP71A1, SLCYP71A1-like, SLCYP85A3, SLCYP734A7, and SLCYP734A8 genes were confirmed through RT-PCR. The changed seed size of a transgenic tomato plant on the influence of citrus cuCYP71A1 gene. At the time of infecting the citrus cuCYP71A1 gene into to Micro-Tom tomato, the SLCYP78A3 gene was suppressed and the decrease of the transgenic tomato seed size of the overexpressed sense line. Analysis of tomato total flavonoid content, total polyphenol content, and antioxidant activity (DPPH) was confirmed. As a result, it was confirmed that the total flavonoid content overexpression sense line was twice higher than the knock-out AS line and controlMT line in fruit. There was no difference between leaves and flowers. Total polyphenol content and antioxidant activity are confirmed was no difference in all stages and lines. The sense line in the total flavonoid content experiment was high in the fruit, the expression level of flavonoid-related genes in the tomato flavonoid pathway was confirmed through RT-PCR analysis. The result was confirmed that there was no difference in the amount of expression. As described above, the High-Performance Liquid Chromatography experiment was confirmed by flavonoid analysis. The result was similar to the result of the total flavonoid content experiment, the sense line was twice higher than anti-sense, and control MT in the fruit. It was also confirmed that there was no difference between leaves and flowers.