Pterostilbene production of tomato transformed with resveratrol synthase and resveratrol O-methyltransferase genes
2022
Zhang, Yue | Ni, Lei | Chen, Shihao | Qin, Yafei | Ding, Xing | Li, Jinhua | Pan, Yu | Zhang, Xingguo
Pterostilbene is a methylated derivative of resveratrol. It has been proved to be effective in preventing many human diseases. However, it is produced and accumulated in only small amounts in natural plant raw materials. Here, two genes coding for resveratrol synthase 3 (AhRS3) in Arachis hypogaea and resveratrol O-methyltransferase (VvROMT) in Vitis vinifera were artificially synthesized considering the codon preference of the tomato. They were linked by LP4/2A to form a fusion gene, controlled by cauliflower mosaic virus 35S promoter, and introduced into tomato via Agrobacterium-mediated transformation. This study aimed to obtain a tomato breeding material enriched with pterostilbene in fruits for a healthy food source. Two transgenic plants with high alien gene expression were selected from the regenerated plants using real-time polymerase chain reaction. High-performance liquid chromatography was used to detect the pterostilbene content in fruits. The highest content reached 146.701 ± 47.771 µg/g dry weight, which was significantly higher than natural levels in all other species tested to date. UPLC-MS/MS was used to analyze the differences in metabolites in fruits between the transgenic and wild-type plants to understand the effect of AhRS3-LP4/2A-VvROMT gene on tomato metabolism. Results showed that the synthesis pathway of stilbenes had little influence on the flavonoid metabolic pathway in tomato fruits.
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