Biological Resource Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup, 56212, Republic of Korea.
Biological Resource Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Jeongeup, 56212, Republic of Korea; Department of Pharmaceutical Engineering, Daegu Haany University, Daegu, 38610, Republic of Korea.
Plant Physiol Biochem. 2021 May;162:556-563. doi: 10.1016/j.plaphy.2021.03.015. Epub 2021 Mar 12.
Stilbenes, including resveratrol and viniferins, a small family of polyphenols, are considered the most important phytoalexin group in Vitis species. In a previous study, we found that co-treatment of methyl jasmonate (MJ) and stevioside (STE) resulted in enhanced extracellular production of viniferins in grapevine cell suspension cultures. Thus, to further understand the mechanisms of viniferin production in grapevine cell cultures, we performed transcriptome analysis and isolated seven candidates of grapevine peroxidase genes (VlAPX6, VlGPX5, VlPRX13, VlPRX21, VlPRX35, VlPRX40, and VlPRX50). Bioconversion of trans-resveratrol to δ-viniferin was examined using crude protein extracts isolated from agroinfiltration-based transient expression of VlPRXs in Nicotiana benthamiana. In addition, we found that crude protein extracts from VlPRX21-, VlPRX35-, and VlPRX40-overexpressing (OX) transgenic Arabidopsis plants led to the conversion of trans-resveratrol to δ-viniferin. We found that in vitro experiments with crude protein extracts from VlPRX21-OX and VlPRX35-OX Arabidopsis plants catalyzed the dimerization of trans-resveratrol to δ-viniferin. Our results suggest that VlPRX21 and VlPRX35 encode functional grapevine class III peroxidases and catalyze the oxidative dimerization of trans-resveratrol to form δ-viniferin in grapevine.
芪类化合物，包括白藜芦醇和葡萄素，是一类多酚，被认为是葡萄属植物中最重要的植物抗毒素（phytoalexin）群体。在之前的研究中，我们发现茉莉酸甲酯（methyl jasmonate，MJ）和甜菊苷（stevioside，STE）共同处理会导致葡萄悬浮细胞培养中外源产生的葡萄素增加。因此，为了进一步了解葡萄细胞培养中葡萄素产生的机制，我们进行了转录组分析，并分离出了 7 个葡萄过氧化物酶基因（VlAPX6、VlGPX5、VlPRX13、VlPRX21、VlPRX35、VlPRX40 和 VlPRX50）的候选基因。我们使用从基于农杆菌瞬时表达的烟草原生质体中分离的粗蛋白提取物，对反式白藜芦醇转化为 δ-葡萄素进行了生物转化研究。此外，我们发现，来自 VlPRX21、VlPRX35 和 VlPRX40 过表达（overexpression，OX）转基因拟南芥植物的粗蛋白提取物可将反式白藜芦醇转化为 δ-葡萄素。我们发现，来自 VlPRX21-OX 和 VlPRX35-OX 拟南芥植物粗蛋白提取物的体外实验可催化反式白藜芦醇二聚化为 δ-葡萄素。我们的研究结果表明，VlPRX21 和 VlPRX35 编码具有功能的葡萄属植物类 III 过氧化物酶，并催化反式白藜芦醇的氧化二聚化，形成葡萄中的 δ-葡萄素。
