Key Laboratory of Plant Functional Genomics of the Ministry of Education/Jiangsu Key Laboratory of Crop Genomics and Molecular Breeding/Jiangsu Co-Innovation Center for Modern Production Technology of Grain Crops/Jiangsu Key Laboratory of Crop Genetics and Physiology, College of Agriculture, Yangzhou University, Yangzhou 225009, China.
Department of Biotechnology, School of Life Science and Technology, University of Electronic Science and Technology of China, Chengdu 610054, China.
Int J Mol Sci. 2022 Oct 12;23(20):12166. doi: 10.3390/ijms232012166.
Rice, as a major food crop, provides necessary energy and nutrition for humans and livestock. However, its nutritional value is affected by lysine. Using point mutation, we previously obtained (aspartokinase) and (dihydrodipicolinate synthase) genes insensitive to lysine feedback inhibition and constructed transgenic lines AK2-52 and DHDPS1-22, which show increased lysine synthesis, as well as Ri-12, which shows decreased lysine degradation by inhibiting rice lysine ketoglutarate reductase/saccharopine dehydrogenase (LKR/SDH) activity. In this study, further transgenic lines were hybridized and evaluated. The lysine content of mature seeds from pyramid lines PRD and PRA increased 32.5- and 29.8-fold, respectively, compared with the wild-type, while the three-gene pyramiding line PRDA had a moderate lysine content. The total lysine, total free lysine, and total protein contents of PRD and PRA also increased and had no obvious impact on the physical and chemical quality, seed appearance, and main agronomic traits. Meanwhile, comparative analysis with polygenic polymeric lines GR containing bacterial () and () genes revealed differences in the way bacterial and endogenous rice AK and DHDPS regulate lysine biosynthesis. These results provide a reference for further evaluation and commercialization of high-lysine transgenic rice.
大米是一种主要的粮食作物,为人类和牲畜提供了必要的能量和营养。然而,其营养价值受到赖氨酸的影响。我们之前通过点突变获得了对赖氨酸反馈抑制不敏感的 (天冬氨酸激酶) 和 (二氢吡啶二羧酸合酶) 基因,并构建了 AK2-52 和 DHDPS1-22 转基因系,这些系表现出赖氨酸合成的增加,以及 Ri-12,它通过抑制水稻赖氨酸酮戊二酸还原酶/琥珀酰丙酮脱氢酶 (LKR/SDH) 活性来降低赖氨酸的降解。在这项研究中,进一步杂交和评估了转基因系。与野生型相比,成熟种子的赖氨酸含量分别增加了 32.5 倍和 29.8 倍,而三基因叠加系 PRDA 则具有中等的赖氨酸含量。PRD 和 PRA 的总赖氨酸、总游离赖氨酸和总蛋白含量也增加,对物理和化学质量、种子外观和主要农艺性状没有明显影响。同时,与含有细菌 (天冬氨酸激酶) 和 (二氢吡啶二羧酸合酶) 基因的多基因聚合系 GR 进行比较分析表明,细菌和内源性水稻 AK 和 DHDPS 调节赖氨酸生物合成的方式存在差异。这些结果为进一步评估和商业化高赖氨酸转基因水稻提供了参考。