Table_13_Analysis of chemotypes and their markers in leaves of core collections of Eucommia ulmoides using metabolomics.xlsx

The leaves of Eucommia ulmoides contain various active compunds and nutritional components, and have successively been included as raw materials in the Chinese Pharmacopoeia, the Health Food Raw Material Catalogue, and the Feed Raw Material Catalogue. Core collections of E. ulmoides had been constructed from the conserved germplasm resources basing on molecular markers and morphological traits, however, the metabolite diversity and variation in this core population were little understood. Metabolite profiles of E. ulmoides leaves of 193 core collections were comprehensively characterized by GC-MS and LC-MS/MS based non-targeted metabolomics in present study. Totally 1,100 metabolites were identified and that belonged to 18 categories, and contained 120 active ingredients for traditional Chinese medicine (TCM) and 85 disease-resistant metabolites. Four leaf chemotypes of the core collections were established by integrated uses of unsupervised self-organizing map (SOM), supervised orthogonal partial least squares discriminant analysis (OPLS-DA) and random forest (RF) statistical methods, 30, 23, 43, and 23 chemomarkers were screened corresponding to the four chemotypes, respectively. The morphological markers for the chemotypes were obtained by weighted gene co-expression network analysis (WGCNA) between the chenomarkers and the morphological traits, with leaf length (LL), chlorophyll reference value (CRV), leaf dentate height (LDH), and leaf thickness (LT) corresponding to chemotypes I, II, III, and IV, respectively. Contents of quercetin-3-O-pentosidine, isoquercitrin were closely correlated to LL, leaf area (LA), and leaf perimeter (LP), suggesting the quercetin derivatives might influence the growth and development of E. ulmoides leaf shape.杜仲（Eucommia ulmoides）叶片含有多种活性成分与营养组分，先后被收录为原料纳入《中国药典》、《保健食品原料目录》及《饲料原料目录》。此前研究已基于分子标记与形态性状，从保存的种质资源中构建了杜仲核心种质，但该核心群体的代谢物多样性与变异特征却鲜有研究。本研究通过基于气相色谱-质谱联用（GC-MS）与液相色谱-串联质谱（LC-MS/MS）的非靶向代谢组学技术，对193份核心种质的杜仲叶片代谢物谱进行了全面解析。本研究共鉴定出1100种代谢物，分属18个类别，其中包含120种中药（TCM）活性成分与85种抗病代谢物。本研究整合无监督自组织映射（SOM）、有监督正交偏最小二乘判别分析（OPLS-DA）与随机森林（RF）等统计方法，将核心种质划分为4种叶片化学型，并分别为这4种化学型筛选出30、23、43和23个化学标记物。本研究通过加权基因共表达网络分析（WGCNA）对化学标记物与形态性状进行关联分析，得到对应各化学型的形态标记：叶长（LL）、叶绿素参考值（CRV）、叶齿高度（LDH）以及叶厚（LT）分别对应化学型I、II、III与IV。槲皮素-3-O-戊糖苷（quercetin-3-O-pentosidine）与异槲皮苷（isoquercitrin）的含量与叶长、叶面积（LA）以及叶周长（LP）显著相关，提示槲皮素类衍生物可能影响杜仲叶片形状的生长与发育。