Xu Xiao-shuang, Xue Ying, Zhang Fu-sheng, Qin Xue-mei, Peng Bing, Tian Hong-ling, Zeng Zu-ping
Yao Xue Xue Bao. 2016 Jul;51(7):1165-74.
The content changes of chemical components in different phenological phase of the cultivated Polygala tenuifolia is one of the important factors for determination of the best harvest time in the production practice. In this study, the digital gene expression (DGE) profiles of the cultivated P. tenuifolia were analyzed in different phenological phase (flowering fruit bearing stage, wilting stage, dormancy stage). The differentially expressed genes were found in the biosynthesis of chemical composition in P. tenuifolia, and the representational ones were validated by RT-q PCR. Then, the key enzymes(CYP450s and UGTs) involved in the downstream of the triterpenoid saponins biosynthesis pathway in P. tenuifolia were predicted through the correlation analysis of gene expression. The number of down-regulated genes was more than that of up-regulated in P. tenuifolia from flowering fruit bearing stage to dormancy stage. Six differentially expressed genes (HMGS, PMK, FPPS, SQS, SE, β-AS) and five (PAL, C4 H, 4CL, CAD, peroxidase) were annotated to the triterpenoid saponins and phenylpropanoid biosynthesis pathway in P. tenuifolia, respectively. Compared to wilting and dormancy stages, the saponins, xanthones, and lignins were largely synthesized at the flowering fruit bearing stage of P. tenuifolia. Furthermore, UGT83A1, CYP716B1, CYP98A3, CYP86B1, and CYP94A1 may be the part of key enzymes in the downstream of the triterpenoid saponins biosynthesis pathway in P. tenuifolia. This study provides evidence to support the correctness of traditional harvest time of P. tenuifolia at the level of transcription, and lays the scientific foundation for gene cloning and functional verification of CYP450 s and UGTs in the downstream of the triterpenoid saponins biosynthesis pathway in P. tenuifolia in the future.
栽培远志不同物候期化学成分的含量变化是生产实践中确定最佳采收期的重要因素之一。本研究分析了栽培远志在不同物候期(花果期、枯萎期、休眠期)的数字基因表达(DGE)谱。在远志化学成分的生物合成中发现了差异表达基因,并通过RT-q PCR对代表性基因进行了验证。然后通过基因表达的相关性分析,预测了远志三萜皂苷生物合成途径下游涉及的关键酶(CYP450s和UGTs)。远志从花果期到休眠期下调基因的数量多于上调基因。六个差异表达基因(HMGS、PMK、FPPS、SQS、SE、β-AS)和五个(PAL、C4H、4CL、CAD、过氧化物酶)分别被注释到远志的三萜皂苷和苯丙烷生物合成途径中。与枯萎期和休眠期相比远志的皂苷、呫吨酮和木质素在花果期大量合成。此外,UGT83A1、CYP716B1、CYP98A3、CYP86B1和CYP94A1可能是远志三萜皂苷生物合成途径下游关键酶的一部分。本研究为远志传统采收期在转录水平的正确性提供了证据,并为今后远志三萜皂苷生物合成途径下游CYP450s和UGTs的基因克隆和功能验证奠定了科学基础。