School of Pharmacy, Anhui University of Chinese Medicine, Hefei, 230012, China.
Institute of Traditional Chinese Medicine Resources Protection and Development, Anhui Academy of Chinese Medicine, Hefei, 230012, China.
Sci Rep. 2021 May 7;11(1):9810. doi: 10.1038/s41598-021-89294-1.
Platycodin D and platycoside E are two triterpenoid saponins in Platycodon grandiflorus, differing only by two glycosyl groups structurally. Studies have shown β-Glucosidase from bacteria can convert platycoside E to platycodin D, indicating the potential existence of similar enzymes in P. grandiflorus. An L(3) orthogonal experiment was performed to establish a protocol for calli induction as follows: the optimal explant is stems with nodes and the optimum medium formula is MS + NAA 1.0 mg/L + 6-BA 0.5 mg/L to obtain callus for experimental use. The platycodin D, platycoside E and total polysaccharides content between callus and plant organs varied wildly. Platycodin D and total polysaccharide content of calli was found higher than that of leaves. While, platycoside E and total polysaccharide content of calli was found lower than that of leaves. Associating platycodin D and platycoside E content with the expression level of genes involved in triterpenoid saponin biosynthesis between calli and leaves, three contigs were screened as putative sequences of β-Glucosidase gene converting platycoside E to platycodin D. Besides, we inferred that some transcription factors can regulate the expression of key enzymes involved in triterpernoid saponins and polysaccharides biosynthesis pathway of P. grandiflorus. Totally, a candidate gene encoding enzyme involved in converting platycoside E to platycodin D, and putative genes involved in polysaccharide synthesis in P. grandiflorus had been identified. This study will help uncover the molecular mechanism of triterpenoid saponins biosynthesis in P. grandiflorus.
桔梗中的远志酸 D 和远志酸 E 是两种三萜皂苷,仅在结构上相差两个糖基。研究表明,细菌中的β-葡萄糖苷酶可以将远志酸 E 转化为远志酸 D,表明桔梗中可能存在类似的酶。采用 L(3)正交试验设计,建立了愈伤组织诱导的方案:最佳外植体为带节的茎,最佳培养基配方为 MS+NAA1.0mg/L+6-BA0.5mg/L,以获得实验用愈伤组织。愈伤组织与植物器官之间的远志酸 D、远志酸 E 和总多糖含量差异很大。发现愈伤组织中的远志酸 D 和总多糖含量高于叶片,而远志酸 E 和总多糖含量低于叶片。将愈伤组织和叶片中三萜皂苷生物合成相关基因的表达水平与远志酸 D 和远志酸 E 含量相关联,筛选出三个假定序列为β-葡萄糖苷酶基因,该酶能将远志酸 E 转化为远志酸 D。此外,我们推断出一些转录因子可以调节三萜皂苷和多糖生物合成途径中关键酶的表达。总的来说,鉴定了一个编码将远志酸 E 转化为远志酸 D 的酶的候选基因,以及一个参与桔梗多糖合成的假定基因。本研究将有助于揭示桔梗中三萜皂苷生物合成的分子机制。