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转白藜芦醇合酶和白藜芦醇 O-甲基转移酶基因的番茄中天南星素的生产。

Pterostilbene production of tomato transformed with resveratrol synthase and resveratrol O-methyltransferase genes.

机构信息

College of Horticulture and Landscape Architecture, Southwest University, No.2 Tiansheng Road, Beibei, Chongqing 400715, China; Key Laboratory of Horticulture Science for Southern Mountainous Regions, the Ministry of Education, Southwest University, No.2 Tiansheng Road, Beibei, Chongqing 400715, China; State Cultivation Base of Crop Stress Biology for Southern Mountainous Land of Southwest University, Academy of Agricultural Sciences, Southwest University, Beibei, Chongqing 400715, China; Department of Agriculture and Animal Husbandry, Aba Vocational College, No. 252, South Section of Fengyi Avenue, Fengyi Town, Maoxian County, Aba Tibetan and Qiang Autonomous Prefecture, Sichuan 61002500, China.

College of Horticulture and Landscape Architecture, Southwest University, No.2 Tiansheng Road, Beibei, Chongqing 400715, China; Key Laboratory of Horticulture Science for Southern Mountainous Regions, the Ministry of Education, Southwest University, No.2 Tiansheng Road, Beibei, Chongqing 400715, China; State Cultivation Base of Crop Stress Biology for Southern Mountainous Land of Southwest University, Academy of Agricultural Sciences, Southwest University, Beibei, Chongqing 400715, China.

出版信息

Plant Sci. 2022 Sep;322:111343. doi: 10.1016/j.plantsci.2022.111343. Epub 2022 Jun 1.

Abstract

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.

摘要

紫檀芪是白藜芦醇的甲基化衍生物。已证明它在预防许多人类疾病方面非常有效。然而,它在天然植物原料中的产量和积累量都非常少。在这里,考虑到番茄的密码子偏好,人工合成了花生中的白藜芦醇合酶 3(AhRS3)和葡萄中的白藜芦醇 O-甲基转移酶(VvROMT)这两个基因。它们通过 LP4/2A 连接在一起形成融合基因,由花椰菜花叶病毒 35S 启动子控制,并通过根癌农杆菌介导的转化导入番茄。本研究旨在获得富含紫檀芪的果实番茄育种材料,作为健康食品的来源。从再生植株中使用实时聚合酶链反应选择了两个具有高异源基因表达的转基因植株。使用高效液相色谱法检测果实中的紫檀芪含量。最高含量达到 146.701±47.771µg/g 干重,明显高于迄今为止测试的所有其他物种的天然水平。使用 UPLC-MS/MS 分析转基因和野生型植物果实中代谢物的差异,以了解 AhRS3-LP4/2A-VvROMT 基因对番茄代谢的影响。结果表明,芪类化合物的合成途径对番茄果实中类黄酮代谢途径的影响很小。

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