Kai Hou

Heavy-metal ATPase (HMA) is widely involved in the absorption and transport of metal elements, including copper, zinc, and cadmium in plants. Although the characterization of HMA has been conducted in several plants, the research on the HMA gene in Stevia rebaudiana remains limited. In this study, twelve SrHMA genes were identified, encoding stable proteins mostly localized to the plasma membrane and possessing conserved E1-E2_ATPase and hydrolase domains, with one to three N-terminal HMA domains. Phylogenetic analysis classified them into two subfamilies (Zn/Co/Cd/Pb and Cu/Ag P1B-ATPases). The genes are unevenly distributed across five chromosomes, with two segmental duplication events and an independent genome duplication event specific to dicots, and their promoters harbor abundant stress-responsive elements. Expression profiling under copper and cadmium stresses showed distinct tissue specificity: cadmium stress upregulated most genes in roots but suppressed them in leaves, whereas copper stress upregulated all genes in leaves and half of the genes in roots. This study represents the first systematic analysis of the SrHMA gene family, revealing its potential involvement in heavy-metal stress responses and providing candidate genes for future functional characterization and breeding of S. rebaudiana varieties with enhanced metal tolerance.

Houttuynia cordata is a traditional medicinal plant with various bioactivities mainly caused by its terpenoids. However, the effects of geography, environment, biosynthesis, and multiple pharmacological targets of these compounds are still not well understood . We used UPLC‐MS/MS and GC–MS metabolomics to analyze terpenoids in accession 7# grown across six regions in China (Yunnan, Guangxi, Hubei, Chongqing, Guizhou, and Sichuan) and identified key environmental factors linked through random forest analysis. Network pharmacology identified potential targets and pathways for differentially accumulated terpenoids, further validated by molecular docking. Transcriptome sequencing identified terpenoid biosynthetic genes. A total of 502 terpenoid metabolites were detected, and chemotypic diversity was strongly shaped by geographical origin. Altitude (bio21) and annual precipitation (bio12) emerged as the primary environmental factors associated with 58 and 18 differential metabolites, respectively. Network analysis of 39 terpenoids revealed 239 potential targets, with 23 core targets (e.g., ESR1, STAT3, BCL2, AR) enriched in cancer, endocrine resistance, and hormone‐related pathways. Docking confirmed stable interactions between key terpenoids (byzantionoside B, ursonic Acid) and core targets (ESR1, AR, BCL2) with binding energies < −7.5 kcal mol ⁻¹ . Transcriptomic analysis uncovered 103 differentially expressed genes in the MVA and MEP pathways, with several (e.g., AACT, FPPS, HMGR, DXS) showing strong correlations with core terpenoid accumulation. This multi‐omics study provides insights into substantial geospatial variation in H. cordata terpenoids, identifies altitude and precipitation as major environmental factors associated with terpenoid accumulation, and offers a predictive framework for the biosynthetic network and multi‐target pharmacological potential, especially anticancer. It offers a theoretical basis for quality control and optimized cultivation.

This study examines Chuan Xiong (川芎Rhizoma Chuanxiong), a traditional Chinese medicinal herb cultivated in Sichuan province, China. By systematically investigating its botanical origin, cultivation history, and international trade dynamics, the study identifies a sharp decline in global exports—once peaking at approximately 10,000 tons—and evaluates the impact of divergent pharmacopoeial standards, particularly the 2017 requirements of the European Pharmacopoeia , on trade performance. Through a historical literature review and a comparative analysis of pharmacopoeial standards across different countries and regions, the research confirms the Sichuan-specific origin and cultivation heritage of Chuan Xiong , and reveals inconsistencies in key quality control parameters, including botanical identification, macroscopic characteristics, moisture limits, and bioactive constituents. Through iterative negotiations and scientific consultations among relevant stakeholders, an international consensus was achieved, resulting in the development of a harmonized, evidence-based quality framework which was subsequently adopted as an ISO standard under ISO/TC 249. This internationally aligned standard addresses previous discrepancies in both parameter selection and specification limits, more accurately reflecting the distinctive quality profile of Chuan Xiong . The establishment of this ISO standard provides a scientifically robust and globally recognized basis for quality control, and is expected to support the recovery of exports and facilitate the integration of Chuan Xiong into international markets. Ultimately, this initiative provides a replicable model for the global harmonization of quality standards for traditional medicinal botanicals.

Stevia rebaudiana Bertoni is recognized as the third major source of natural sweetness due to its leaves being rich in high-sweetness, low-calorie steviol

Background: Morus alba L. (family Moraceae) is widely cultivated across the world and is well-known for its medicinal and nutritional value, especially its leaves. This study investigates the regional variation in mulberry leaf metabolites, focusing on alkaloids and flavonoids, and explores the influence of climatic and environmental factors on their biosynthesis using an integrated metabolomic and environmental analysis. Mulberry leaves, known for their medicinal and nutritional value, were collected from six regions across China, including Sichuan, Xinjiang, and Tibet. Methods: Untargeted metabolomics via UHPLC-MS was conducted. Differential metabolites were identified through multivariate analysis and annotated using the KEGG database. Redundancy analysis was used to link metabolite profiles with climatic data. Results: Mulberry leaves from six Chinese regions showed significant variation in total flavonoid content (TFC), total polyphenol content (TPC), and 1-Deoxynojirmycin (DNJ), with Tibet having the highest TFC and TPC, and Panzhihua the highest DNJ. Metabolomic analysis identified 3794 metabolites, revealing distinct regional clustering. A total of 79 differential metabolites were identified, which are enriched in pathways such as galactose metabolism and phenylalanine biosynthesis. Environmental factors, especially bio3, bio10, bio2, bio5, and bio20, strongly influenced metabolite profiles. Conclusions: The biosynthesis and accumulation of secondary metabolites in mulberry leaves are significantly influenced by region-specific environmental factors, particularly temperature, precipitation, and light. The identified differential metabolites are mainly enriched in galactose metabolism, arginine, and proline metabolism, and phenylalanine, tyrosine, and tryptophan biosynthesis. These pathways are closely associated with plant stress responses and the synthesis of secondary metabolites. The pronounced regional differences in metabolite profiles underscore the critical role of environmental factors in determining the chemical composition of mulberry leaves. This research provides valuable insights into the influence of climatic factors affecting the chemical composition of plants. It lays a theoretical foundation for the quality assessment and grading of mulberry leaves, providing scientific guidance for their targeted cultivation and utilization.

With growing public concern about the health effects of low-dose radiation, numerous studies have demonstrated that low-dose radiation can cause damage to the immune system, making intervention measures essential. This study investigated the protective effects of silybin against low-dose radiation-induced immune system damage and its underlying mechanisms at both the cellular and animal levels. At the cellular level, CCK-8 assays, ROS measurements, and RT-qPCR analysis revealed that silybin alleviated the reduction in RAW264.7 cell proliferation, intracellular ROS levels, and inflammatory cytokine expression following low-dose radiation exposure. At the animal level, comparative analyses of post-irradiation body weight, peripheral blood cell counts, immune organ coefficients, spleen HE/IHC staining, and spleen immune cell numbers demonstrated that silybin mitigated the radiation-induced decrease in body weight, reduction in peripheral blood leukocyte counts, inflammatory cell infiltration in the spleen, decline in spleen immune cell numbers, and increase in cGAS protein-positive cells. These findings indicate that silybin exerts protective effects against low-dose radiation-induced immune system damage, potentially by regulating the cGAS signaling pathway to reduce radiation-induced cellular injury, thereby enhancing its radioprotective properties.

Objective: In Tibetan dietary and folk medicine practices, Rheum austral is commonly used as an alternative to Rheum tanguticum, and there is a prevailing belief that wild rhubarb should not be substituted by its cultivated counterpart. However, these traditions are not supported by scientific evidence, particularly concerning the differences in endogenous metabolites between cultivated and wild rhubarbs, as well as between officially recognized and non-official rhubarbs. These uncertainties have also been hindering the vertical integration development of the local rhubarb industry.Methods: In this study, ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-QTOFMS) and biostatistical analysis were employed to systematically and comprehensively investigate the chemical constituents of rhubarbs from various sources, focusing on the differences in metabolic components between cultivated and wild rhubarbs.Results: The metabolic differences in rhubarb from various varieties and environments are pronounced. Among them, 39 differential metabolites were identified between cultivated R. tanguticum and wild R. tanguticum. cultivated R. tanguticum is rich in emodin, physcion, and rhapontigenin, whereas wild R. tanguticum exhibits a higher concentration of rhaponticin and is particularly abundant in anthraquinone compounds. Additionally, 33 differential metabolites distinguished wild R. tanguticum from wild R. austral, with R. austral being rich in stilbene derivatives and wild R. tanguticum predominantly containing coumarins. The correlations among these differential metabolites have also been further explored and presented.Conclusion: The metabolic disparities between cultivated and wild rhubarb varieties are substantial, with wild rhuabarb containing higher levels of effective components than its cultivated counterparts. However, wild varieties face issues with component instability and resource depletion, while cultivated varieties exhibit more stable effective components. Given these significant differences in metabolic components, it is essential to differentiate rhubarbs from various species and growing conditions to suit specific medicinal and dietary purposes effectively. This paper can lay a theoretical foundation for the vertical integration development of the rhubarb industry in Tibetan areas.

Angelica dahurica var. formosana (ADF), which belongs to the Umbelliferae family, is one of the original plants of herbal raw material Angelicae Dahuricae Radix. ADF roots represent an enormous biomass resource convertible for disease treatment and bioproducts. But, early bolting of ADF resulted in lignification and a decrease in the coumarin content in the root, and roots lignification restricts its coumarin for commercial utility. Although there have been attempts to regulate the synthesis ratio of lignin and coumarin through biotechnology to increase the coumarin content in ADF and further enhance its commercial value, optimizing the biosynthesis of lignin and coumarin remains challenging. Based on gene expression analysis and phylogenetic tree profiling, AdNAC20 as the target for genetic engineering of lignin and coumarin biosynthesis in ADF was selected in this study. Early-bolting ADF had significantly greater degrees of root lignification and lower coumarin contents than that of the normal plants. In this study, overexpression of AdNAC20 gene plants were created using transgenic technology, while independent homozygous transgenic lines with precise site mutation of AdNAC20 were created using CRISPR/Cas9 technology. The overexpressing transgenic ADF plants showed a 9.28% decrease in total coumarin content and a significant 12.28% increase in lignin content, while knockout mutant plants showed a 16.3% increase in total coumarin content and a 33.48% decrease in lignin content. Furthermore, 29,671 differentially expressed genes (DEGs) were obtained by comparative transcriptomics of OE-NAC20, KO-NAC20, and WT of ADF. A schematic diagram of the gene network interacting with AdNAC20 during the early-bolting process of ADF was constructed by DEG analysis. AdNAC20 was predicted to directly regulate the transcription of several genes with SNBE-like motifs in their promoter, such as MYB46, C3H, and CCoAOMT. In this study, AdNAC20 was shown to play a dual pathway function that positively enhanced lignin formation but negatively controlled coumarin formation. And the heterologous expression of the AdNAC20 gene at Arabidopsis thaliana proved that the AdNAC20 gene also plays an important role in the process of bolting and flowering.

The basic (region) leucine zippers (bZIPs) are evolutionarily conserved transcription factors widely distributed in eukaryotic organisms. In plants, they are not only involved in growth and development, defense and stress responses and regulation of physiological processes but also play a pivotal role in regulating secondary metabolism. To explore the function related to the bZIP gene family in Stevia rebaudiana Bertoni, we identified 105 SrbZIP genes at the genome-wide level and classified them into 12 subfamilies using bioinformation methods. Three main classes of cis-acting elements were found in the SrbZIP promoter regions, including development-related elements, defense and stress-responsive elements and phytohormone-responsive elements. Through protein–protein interaction network of 105 SrbZIP proteins, SrbZIP proteins were mainly classified into four major categories: ABF2/ABF4/ABI5 (SrbZIP51/SrbZIP38/SrbZIP7), involved in phytohormone signaling, GBF1/GBF3/GBF4 (SrbZIP29/SrbZIP63/SrbZIP60) involved in environmental signaling, AREB3 (SrbZIP88), PAN (SrbZIP12), TGA1 (SrbZIP69), TGA4 (SrbZIP82), TGA7 (SrbZIP31), TGA9 (SrbZIP95), TGA10 (SrbZIP79) and HY5 (SrbZIP96) involved in cryptochrome signaling, and FD (SrbZIP72) promoted flowering. The transcriptomic data showed that SrbZIP genes were differentially expressed in six S. rebaudiana cultivars (‘023’, ‘110’, ‘B1188’, ‘11-14’, ‘GP’ and ‘GX’). Moreover, the expression levels of selected 15 SrbZIP genes in response to light, abiotic stress (low temperature, salt and drought), phytohormones (methyl jasmonate, gibberellic acid and salicylic acid) treatment and in different tissues were analyzed utilizing qRT-PCR. Some SrbZIP genes were further identified to be highly induced by factors affecting glycoside synthesis. Among them, three SrbZIP genes (SrbZIP54, SrbZIP63 and SrbZIP32) were predicted to be related to stress-responsive terpenoid synthesis in S. rebaudiana. The protein–protein interaction network expanded the potential functions of SrbZIP genes. This study firstly provided the comprehensive genome-wide report of the SrbZIP gene family, laying a foundation for further research on the evolution, function and regulatory role of the bZIP gene family in terpenoid synthesis in S. rebaudiana.