Hou Jiaqi, Zhou Yuhan, Ran Liping, Chen Yanzhu, Zhang Ting, Sun Bowei, Yang Yimo, Sang Qianzi, Cao Li
Agriculture College, Yanbian University, Yanji 133002, China.
Metabolites. 2023 Feb 1;13(2):216. doi: 10.3390/metabo13020216.
L. (HSK) is a rare and endangered species in the wild that grows on the cliffs of deep mountains. As a natural plant, the chemical composition of HSK is relatively complex, which mainly includes flavonoids, organic acids, polysaccharides, and various trace elements with good effects of clearing away heat, anti-inflammatory, analgesic, and calming nerves, and inhibiting tumor cells. In this experiment, different developmental stages of HSK flowers were used for optimization of the flavonoid extraction and determining method. The antioxidant activities, flavonoid accumulation pattern, and synthesis regulatory network were analyzed using biochemistry, RNA-seq, and UPLC-MS/MS. The total content of flavonoids, vitexin rhamnoside, hyperoside, and rutin in HSK flowers at T3 stage (flower wilting) was significantly higher than in T2 (full flowering) and T1 (bud) stages. Compared with T1 and T2, the antioxidant capacity of the T3 flower alcohol extract was also the strongest, including the total reducing ability, DPPH clearance, OH clearance, O clearance, and total antioxidant capacity. A total of 156 flavonoids and 47,179 unigenes were detected by UPLC-MS/MS and RNA-Seq, respectively. The candidate genes and key metabolites involved in flavonoid biosynthesis were identified and the regulatory networks were also analyzed in this study. qRT-PCR test further proved that the gene expression level was consistent with the results of RNA sequence data. The relationship between the gene expression and flavonoid accumulation network provides a theoretical basis for the mining and regulation of functional genes related to the flavonoid biosynthesis and metabolism in L.
石蒜（HSK）是一种生长在深山悬崖上的野生珍稀濒危物种。作为一种天然植物，石蒜的化学成分相对复杂，主要包括黄酮类化合物、有机酸、多糖以及各种微量元素，具有清热、抗炎、镇痛、安神和抑制肿瘤细胞等良好功效。在本实验中，利用石蒜花的不同发育阶段对黄酮类化合物的提取及测定方法进行优化。采用生物化学、RNA测序和超高效液相色谱-串联质谱法分析其抗氧化活性、黄酮类化合物积累模式及合成调控网络。在T3阶段（花枯萎期）石蒜花中黄酮类化合物、牡荆素鼠李糖苷、金丝桃苷和芦丁的总含量显著高于T2阶段（盛花期）和T1阶段（花蕾期）。与T1和T2阶段相比，T3阶段花的醇提取物抗氧化能力也最强，包括总还原能力、DPPH清除率、OH清除率、O清除率和总抗氧化能力。通过超高效液相色谱-串联质谱法和RNA测序分别检测到156种黄酮类化合物和47179个单基因。本研究鉴定了参与黄酮类生物合成的候选基因和关键代谢产物，并分析了调控网络。qRT-PCR检测进一步证明基因表达水平与RNA序列数据结果一致。基因表达与黄酮类化合物积累网络之间的关系为石蒜中黄酮类生物合成与代谢相关功能基因的挖掘和调控提供了理论依据。
