Flower Research Institute of Yunnan Academy of Agricultural Sciences, Key Lab of Yunnan Flower Breeding, National Engineering Research Center For Ornamental Horticulture, Kunming 650205, China.
Genes (Basel). 2023 Oct 9;14(10):1920. doi: 10.3390/genes14101920.
Flavonoid-3',5'-hydroxylase (F3'5'H) is the key enzyme for the biosynthesis of delphinidin-based anthocyanins, which are generally required for purple or blue flowers. Previously, we isolated a full-length cDNA of from , which shared the highest homology with . In this study, was subcloned into a plant over-expression vector and transformed into tobacco via to investigate its catalytic function. Positive transgenic tobacco T plants were obtained by hygromycin resistance screening and PCR detection. showed a higher expression level in all transgenic tobacco plants than in control plants. Under the drive of the cauliflower mosaic virus (CaMV) 35S promoter, the over-expressed produced dihydromyricetin (DHM) and some new anthocyanin pigments (including delphinidin, petunidin, peonidin, and malvidin derivatives), and increased dihydrokaempferol (DHK), taxifolin, tridactyl, cyanidin derivatives, and pelargonidin derivatives in transgenic tobacco plants by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) analysis, resulting in a dramatic color alteration from light pink to magenta. These results indicate that products have F3'5'H enzyme activity. In addition, transfer alters flavonoid pigment synthesis and accumulation in tobacco. Thus, may be considered a candidate gene for gene engineering to enhance anthocyanin accumulation and the molecular breeding project for blue flowers.
类黄酮 3',5'-羟化酶（F3'5'H）是花色苷生物合成的关键酶，花色苷通常是紫色或蓝色花所必需的。此前，我们从 中分离出全长 cDNA ，其与 具有最高同源性。在这项研究中， 将被亚克隆到植物过表达载体中，并通过 转化到烟草中，以研究其催化功能。通过潮霉素抗性筛选和 PCR 检测获得了阳性转基因烟草 T 植株。 在所有转基因烟草植株中的表达水平均高于对照植株。在花椰菜花叶病毒（CaMV）35S 启动子的驱动下，过表达的 产生了二氢杨梅素（DHM）和一些新的花色苷色素（包括飞燕草素、矢车菊素、锦葵素和翠雀素衍生物），并通过超高效液相色谱-串联质谱（UPLC-MS/MS）分析增加了二氢山奈酚（DHK）、杨梅素、三叶苷、矢车菊素衍生物和天竺葵素衍生物在转基因烟草植株中的含量，导致颜色从浅粉色变为洋红色。这些结果表明 具有 F3'5'H 酶活性。此外， 的转移改变了烟草中类黄酮色素的合成和积累。因此， 可能被认为是增强花色苷积累的基因工程和蓝色花卉分子育种项目的候选基因。
