Key Laboratory of Biological Molecular Medicine Research of Guangxi Higher Education, Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Guangxi Medical University, Nanning, Guangxi province, China.
Department of Nursing, Sichuan Nursing Vocational College, Sichuan province, China.
PLoS One. 2021 Dec 7;16(12):e0260027. doi: 10.1371/journal.pone.0260027. eCollection 2021.
Previous studies have revealed that gypenosides produced from Gynostemma pentaphyllum (Thunb.) Makino are mainly dammarane-type triterpenoid saponins with diverse structures and important biological activities, but the mechanism of diversity for gypenoside biosynthesis is still unclear. In this study, a combination of isobaric tags for relative and absolute quantification (iTRAQ) proteome analysis and RNA sequencing transcriptome analysis was performed to identify the proteins and genes related to gypenoside biosynthesis. A total of 3925 proteins were identified by proteomic sequencing, of which 2537 were quantified. Seventeen cytochrome P450 (CYP) and 11 uridine 5'-diphospho-glucuronosyltransferase (UDP-glucuronosyltransferase, UGT) candidate genes involved in the side chain synthesis and modification of gypenosides were found. Seven putative CYPs (CYP71B19, CYP77A3, CYP86A7, CYP86A8, CYP89A2, CYP90A1, CYP94A1) and five putative UGTs (UGT73B4, UGT76B1, UGT74F2, UGT91C1 and UGT91A1) were selected as candidate structural modifiers of triterpenoid saponins, which were cloned for gene expression analysis. Comprehensive analysis of RNA sequencing and proteome sequencing showed that some CYPs and UGTs were found at both the transcription and translation levels. In this study, an expression analysis of 7 CYPs and 5 UGTs that contributed to gypenoside biosynthesis and distribution in G. pentaphyllum was performed, providing consistent results that will inspire more future research on vital genes/proteins involved in gypenoside biosynthesis.
先前的研究表明，绞股蓝中产生的绞股蓝皂苷主要是达玛烷型三萜皂苷，具有多种结构和重要的生物活性，但苷元生物合成的多样性机制尚不清楚。在这项研究中，采用相对和绝对定量同位素标记（iTRAQ）蛋白质组分析和 RNA 测序转录组分析相结合的方法，鉴定与绞股蓝皂苷生物合成相关的蛋白质和基因。通过蛋白质组测序共鉴定到 3925 种蛋白质，其中 2537 种蛋白质被定量。发现了 17 种细胞色素 P450（CYP）和 11 种尿苷二磷酸葡萄糖醛酸基转移酶（UDP-葡萄糖醛酸基转移酶，UGT）候选基因，这些候选基因参与了绞股蓝皂苷侧链的合成和修饰。鉴定了 7 个假定的 CYP（CYP71B19、CYP77A3、CYP86A7、CYP86A8、CYP89A2、CYP90A1、CYP94A1）和 5 个假定的 UGT（UGT73B4、UGT76B1、UGT74F2、UGT91C1 和 UGT91A1）作为三萜皂苷结构修饰的候选基因，并对其进行了基因表达分析。RNA 测序和蛋白质组测序的综合分析表明，一些 CYP 和 UGT 同时在转录和翻译水平上被发现。在这项研究中，对参与绞股蓝皂苷生物合成和分布的 7 个 CYP 和 5 个 UGT 进行了表达分析，结果一致，为参与绞股蓝皂苷生物合成的关键基因/蛋白提供了更多的研究启示。
