Untargeted Metabolomics Reveals Multiple Phytometabolites in the Agricultural Waste Materials and Medicinal Materials of Codonopsis pilosula

Codonopsis pilosula has been used in traditional Chinese medicine for hundreds of years, where it has been used to treat anaemia, fatigue, a weak spleen, and stomach problems, among other ailments. The roots of C. pilosula are considered medicinal, while the aerial parts are always directly discarded after harvest in autumn or winter. Some studies have shown that the stems and leaves of C. pilosula also contain a variety of active metabolites, including saponins, flavonoids, terpenoids, and polysaccharides. To efficiently utilise resources, waste products from C. pilosula leaves and stems were analysed by untargeted metabolomics and chemometrics. A total of 1508 metabolites were detected and annotated, of which 463 were identified as differentially expressed metabolites (DEMs). These DEMs were grouped into classes, such as carboxylic acids and derivatives, steroids, organic oxygen compounds, fatty acyls, prenol lipids, and flavonoids. Metabolic profiling of C. pilosula tissues showed that the contents of polyacetylenes, polyenes, flavonoids, some alkaloids, steroids, terpenoids, and organic acids were higher in stems and leaves, whereas the contents of the main lignans and some alkaloids were more enriched in roots. Moreover, C. pilosula stems and leaves also contained a lobetyolin, syringin and atractylenolide III, which were detected by LC-MS/MS and HPLC-UV. The extracts of C. pilosula aerial parts also showed stronger antioxidant properties than roots. C. pilosula stems and leaves were rich in active ingredients and might have great value for development and utilisation.

Prediction of Potentially Suitable Distributions of Codonopsis pilosula in China Based on an Optimized MaxEnt Model

Species distribution models are widely used in conservation biology and invasive biology. MaxEnt models are the most widely used models among the existing modeling tools. In the MaxEnt modeling process, the default parameters are used most often to build the model. However, these models tend to be overfit. Aiming at this problem, this study uses an optimized MaxEnt model to analyze the impact of past, present and future climate on the distributions of Codonopsis pilosula, an economic species, to provide a theoretical basis for its introduction and cultivation. Based on 264 distribution records and eight environmental variables, the potential distribution areas of C. pilosula in the last interglacial, middle Holocene and current periods and 2050 and 2070 were simulated. Combined with the percentage contribution, permutation importance, and jackknife test, the environmental factors affecting the suitable distribution area of this species were discussed. The results show that the parameters of the optimal model are: the regularization multiplier is 1.5, and the feature combination is LQHP (linear, quadratic, hinge, product). The main temperature factors affecting the distribution of C. pilosula are the annual mean temperature, mean diurnal range, and isothermality. The main precipitation factors are the precipitation seasonality, precipitation in the wettest quarter, and precipitation in the driest quarter, among which the annual average temperature contributes the most to the distribution area of this species. With climate warming, the suitable area of C. pilosula exhibits a northward expansion trend. It is estimated that in 2070, the suitable area of this species will expand to its maximum, reaching 2.5108 million square kilometers. The highly suitable areas of C. pilosula are mainly in Sichuan, Gansu, Shaanxi, Shanxi, and Henan Provinces. Our findings can be used to provide theoretical support related to avoiding the blind introduction of C. pilosula.

Contents of lobetyolin, syringin, and atractylolide III in Codonopsis pilosula are related to dynamic changes of endophytes under drought stress

Background Codonopsis pilosula, an important medicinal plant, can accumulate certain metabolites under moderate drought stress. Endophytes are involved in the metabolite accumulations within medicinal plants. It is still unknown that the endophytes of C. pilosula are associated with the accumulations of metabolites. This study aims to investigate the promoting effect of endophytes on the accumulations of active substances in C. pilosula under drought stress. Methods High–performance liquid chromatography and high–throughput sequencing technology were performed to investigate changes in the contents of secondary metabolite and endophyte abundances of C. pilosula under drought stress, respectively. Spearman’s correlation analysis was further conducted to identify the endophytic biomarkers related to accumulations of pharmacodynamic compounds. Culture-dependent experiments were performed to confirm the functions of endophytes in metabolite accumulations. Results The distribution of pharmacological components and diversity and composition of endophytes showed tissue specificity within C. pilosula. The contents of lobetyolin, syringin, and atractylolide III in C. pilosula under drought stress were increased by 8.47%‒86.47%, 28.78%‒230.98%, and 32.17%‒177.86%, respectively, in comparison with those in untreated groups. The Chao 1 and Shannon indices in different parts of drought–stressed C. pilosula increased compared with those in untreated parts. The composition of endophytic communities in drought treatment parts of C. pilosula was different from that in control parts. A total of 226 microbial taxa were identified as potential biomarkers, of which the abundances of 42 taxa were significantly and positively correlated to the pharmacodynamic contents. Culture-dependent experiments confirmed that the contents of lobetyolin and atractylolide III were increased by the application of Epicoccum thailandicum, Filobasidium magnum, and Paraphoma rhaphiolepidis at the rates of 11.12%‒46.02%, and that the content of syringin was increased by Pseudomonas nitroreducens at the rates of 118.61%‒119.36%. Conclusions Certain endophytes participated in the accumulations of bioactive metabolites, which provided a scientific evidence for the development and application of microorganisms to improve the quality of traditional Chinese medicine.

Screening of Active Components and Key Targets of Radix Codonopsis in the Treatment of Gastric Cancer

Gastric cancer is the fifth most common cancer type in the world. The incidence and mortality of gastric cancer in China ranks second among malignant tumors in the country. At present, the main treatment method of gastric cancer is still surgical resection combined with chemotherapy. However, chemotherapy drugs will cause serious toxic and side effects on other normal tissues and cells. At the same time, chemotherapy drugs can make patients develop drug resistance and seriously affect the curative effect. By contrast, Chinese medicine has more advantages in the treatment of cancer. Dangshen (Radix Codonopsis), a traditional Chinese medicine, has been proved to be effective for the clinical treatment of gastric cancer. However, due to the complex components of Dangshen, the main active components and pharmacological mechanism for its treatment of gastric cancer are still unclear. In this study, the main active components and pharmacological mechanism of Radix Codonopsis in the treatment of gastric cancer were preliminarily explored based on network pharmacology and molecular docking. We obtained bioactive compounds and targets from Radix Codonopsis from the Chinese Medicine System Pharmacology Database (TCMSP) and constructed the active ingredient-target network of Codonopsis pilosula. We then obtained targets related to gastric cancer from the disease database. The common targets of Radix Codonopsis and gastric cancer were the key target of Radix Codonopsis for the treatment of gastric cancer. Then, we used Metascapedatabase to conduct functional enrichment analysis on the key targets of Radix Codonopsis for the treatment of gastric cancer to clarify the mechanism of Radix Codonopsis for the treatment of gastric cancer. We constructed a network to screen the main bioactive compounds and therapeutic targets, assessed the prognostic value of the main target genes by survival analysis, and finally assessed the binding affinity of the main target genes and main bioactive compounds of Radix Codonopsis for the treatment of gastric cancer by molecular docking. The results showed that the main active compounds of Codonopsis pilosula in treating gastric cancer were luteolin and cryptotanshinone, which played a role in the treatment of gastric cancer through the multitarget and multipathway mechanism.

Genome-Wide Characterization of SPL Gene Family in Codonopsis pilosula Reveals the Functions of CpSPL2 and CpSPL10 in Promoting the Accumulation of Secondary Metabolites and Growth of C. pilosula Hairy Root

SQUAMOSA PROMOTER BINDING PROTEIN-LIKE (SPL) transcription factors play critical roles in regulating diverse aspects of plant growth and development, including vegetative phase change, plant architecture, anthocyanin accumulation, lateral root growth, etc. In the present study, 15 SPL genes were identified based on the genome data of Codonopsis pilosula, a well-known medicinal plant. Phylogenetic analysis clustered CpSPLs into eight groups (G1-G8) along with SPLs from Arabidopsis thaliana, Solanum lycopersicum, Oryza sativa and Physcomitrella patens. CpSPLs in the same group share similar gene structure and conserved motif composition. Cis-acting elements responding to light, stress and phytohormone widely exist in their promoter regions. Our qRT-PCR results indicated that 15 CpSPLs were differentially expressed in different tissues (root, stem, leaf, flower and calyx), different developmental periods (1, 2 and 3 months after germination) and various conditions (NaCl, MeJA and ABA treatment). Compared with the control, overexpression of CpSPL2 or CpSPL10 significantly promoted not only the growth of hairy roots, but also the accumulation of total saponins and lobetyolin. Our results established a foundation for further investigation of CpSPLs and provided novel insights into their biological functions. As far as we know, this is the first experimental research on gene function in C. pilosula.