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Forsythiae Fructus (FF) is a widely used folk medicine in China, Japan, and Korea. The distribution of bioactive constituents throughout the fruit segments has rarely been addressed, although mounting evidence suggests that plant secondary metabolites are synthesized and distributed regularly. The phytochemical profiles of three segments of FF (pericarp, stalk and seed) were firstly revealed by liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based quantitative analysis of twenty-one bioactive constituents, including three phenylethanoid glycosides, five lignans, eight flavonoids, and five phenolic acids to explore the spatial distribution of bioactive constituents. Furthermore, the hierarchical clustering analysis (HCA) and one-way analysis of variance (one-way ANOVA) were conducted to visualize and verify the distribution regularity of twenty-one analytes among three segments. The results showed that phytochemical profiles of the three segments were similar, i.e., phenylethanoid glycosides covering the most part were the predominant compounds, followed by lignans, flavonoids and phenolic acids. Nevertheless, the abundance of twenty-one bioactive constituents among three segments was different. Specifically, phenylethanoid glycosides were highly expressed in the seed; lignans were primarily enriched in the stalk; flavonoids were largely concentrated in the pericarp, while the contents of phenolic acids showed no much difference among various segments. The research improves our understanding of distribution patterns for bioactive constituents in FF, and also complements some scientific data for further exploring the quality formation mechanism of FF. [ABSTRACT FROM AUTHOR]

Spatholobi Caulis (SC), the vine stem of Spatholobus suberectus Dunn, is a widely used traditional Chinese medicine (TCM) for the treatment of blood stasis syndrome and related diseases. Xylem and phloem are the main structures of SC and the color of xylem in SC is red brown or brown while the phloem with resin secretions is reddish brown to dark brown. They are alternately arranged in a plurality of concentric or eccentric rings. In order to investigate the distribution patterns of metabolites in xylem and phloem of SC, an analytical method based on UFLC–QTRAP–MS/MS was established for simultaneous determination of 22 constituents including four flavanols, nine isoflavones, two flavonols, two dihydroflavones, one flavanonol, one chalcone, one pterocarpan, one anthocyanidin and one phenolic acid in the samples (xylem and phloem) from Laos. Furthermore, according to the contents of 22 constituents, heat map, principal components analysis (PCA), orthogonal partial least squares discriminant analysis (OPLS–DA) and t–test were used to evaluate the samples and discover the differences between xylem and phloem of SC. The results indicated that the measured ingredients in xylem and phloem were significantly different. To be specific, the contents of flavonoids in xylem were higher than that in phloem, while the content of protocatechuic acid showed a contrary tendency. This study will not only reveal the distribution patterns of metabolites in xylem and phloem of SC but also facilitate further study on their quality formation. [ABSTRACT FROM AUTHOR]

Lonicera japonica flowers (LJF) is a traditional Chinese medicine packed with phenols constituents and widely used in the treatments of various diseases throughout the world. However, there is still very little known on how LJF identifies and resists salt stress. Here in, we systematically investigated the effect of salt on the phenotypic, metabolite, and transcriptomic in LJF. During long term stress (35 days), 1055 differential expression genes (DEGs) involved in the biosynthesis of secondary metabolites were screened through transcriptome analysis, among which the candidate genes and pathways involved in phenols biosynthesis were highlighted; and performed by phylogenetic tree analysis and multiple nucleotide sequence alignment. Ninety compounds were identified and their relative levels were compared between the control and stressed groups based on the LC-MS analysis, Putative biosynthesis networks of phenolic acid and flavonoid were con-structed with structural DEGs. Strikingly, the expression patterns of structural DEGs were mostly consistent with the variations of phenols under salt stress. Notably, the upregulation of UDP-glycosyl transferases under salt stress indicated post-modification of glycosyl transferases may participate in downstream flavonoids synthesis. This study reveals the relationships of the gene regulation and the phenols biosynthesis in LJF under salt stress, paving the way for the use of gene-specific expression to improve the yield of biocomponent. [Display omitted] • The first employ transcriptome to elucidate molecular mechanism regulating secondary metabolites in LJF under salt stress. • 20 DEGs related to phenols synthesis were identified. • Phylogenetic tree analysis and multiple nucleotide sequence alignment were performed on the selected DEGs. • Putative biosynthesis networks of phenols were constructed with DEGs. • The expressions of UGTs were closely related to the variations of downstream flavonoids synthesis. [ABSTRACT FROM AUTHOR]

Apocyni Veneti Folium (AVF) has been raised great interest in the antioxidant properties recently for the preservation of human health. However, little research was found on the integrate metabolites except our previous investigation on the variations of the bioactive constituents. To understand the salt-tolerant mechanisms of the halophyte, metabolomic platform based on ultra-fast liquid chromatography tandem triple time-of-flight mass/mass spectrometer was applied in this study. The results showed that metabolic profiles were separated and differentiated among groups based on multivariate statistical analysis; different metabolites belonged to various chemical classes. Besides, phenylpropanoid pathway and terpenoid biosynthesis were disturbed in all salt-stressed AVF and low salt-treated group appeared to be better than other samples in terms of relative contents (peak areas) of the wide variety of bioactive components and physiological variations of photosynthetic pigments, osmotic homeostasis, lipid peroxidation product and antioxidative enzymes. This study may provide additional insight into the salt-tolerant mechanisms and the quality assessment of AVF in a holistic level based on the plant metabolomics. Image 1 • Metabolomics platform was developed for studying the salt-tolerant mechanisms and quality assessment of the halophyte. • UFLC-Triple TOF-MS/MS analysis combined with multivariate statistical analysis was used to separate and differentiate AVF. • Significant metabolites responsible for discrimination were identified. • Global visualization of pathway changes and proposed flavonoid biosynthesis were performed. [ABSTRACT FROM AUTHOR]

Highlights • A total of 32 metabolites were simultaneously analyzed by triple quadrupole-linear ion trap mass spectrometry in different parts of licorice. • Major biologically active compounds and important amino acids related to abiotic stress were found with remarkably high level in wild licorice. • High content of phenylalanine and compounds in upstream of flavonoid synthesis in cultivated-type suggest the synthesis may differ at the initial stage. • Our findings will provide a novel clue in agricultural breeding of cultivated licorice and facilitate the further study on licorice's quality formation. Abstract Licorice is a famous Chinese medicinal material widely applied worldwide in food and drugs. It possesses a substantial share in the international and domestic markets. While the demand is continuously increasing, wild-type of licorice is gradually disappearing. Its cultivated-type is significantly different compared to its wild-type, especially the bioactive compounds in dried roots and rhizomes (the medicinal parts of wild and cultivated licorice) are responsible for their quality difference. In this study, a total of thirty-two constituents, including seventeen secondary compounds and fifteen primary metabolites, were simultaneously analyzed by UFLC coupled with triple quadrupole-linear ion trap mass spectrometry (UFLC-QTRAP-MS/MS) in different medicinal parts of licorice. Our findings indicated that the content of major bioactive compounds in wild licorice were significantly higher than those in its cultivated-type. The metabolites in rhizomes showed totally different outcomes between wild and cultivated licorice. Remarkably high level of some important amino acids related to abiotic stress (drought and salt) were found in wild licorice. Notably, the high contents of phenylalanine and compounds in upstream of flavonoid synthesis in cultivated licorice suggested that the synthesis may differ at the initial stage. The distribution pattern of metabolites in different medicinal parts of wild and cultivated licorice will not only provide a novel clue in agricultural breeding but also facilitate the further study on their quality formation. [ABSTRACT FROM AUTHOR]

Scrophulariae Radix is one of the most popular traditional Chinese medicines. The harvesting time of Scrophulariae Radix is closely related to the quality of products in this traditional Chinese medicine. The goal of the study is to analyze the dynamic changes of metabolite accumulation in Scrophulariae Radix. The difference of constituents in Scrophulariae Radix harvested at different times was analyzed by liquid chromatography with triple quadrupole-time of flight tandem mass spectrometry coupled with principal component analysis and partial least-square discriminant analysis. According to the accurate mass of molecular and product ions provided by triple quadrupole-time of flight tandem mass spectrometry, a total of 30 differential constituents were identified. Furthermore, the contents of ten index differential constituents in Scrophulariae Radix were simultaneously determined by liquid chromatography coupled with triple quadrupole-linear ion trap tandem mass spectrometry. Gray relational analysis was performed to evaluate the samples harvested at different times according to the contents of ten constituents. All of the results demonstrated that the quality of Scrophulariae Radix collected at traditional harvest time was better. This study will provide the basic information for revealing the dynamic change law of metabolite accumulation of Scrophulariae Radix and exploring its quality forming mechanism. [ABSTRACT FROM AUTHOR]

Pseudostellaria heterophylla is an important traditional Chinese herbal medicine with vast clinical consumption because of its positive effects. To date, changes in the metabolite composition of P. heterophylla have been well documented; however, the molecular differences between cultivated P. heterophylla and its wild-type are still unknown. The aim of this study was to investigate differences in cultivated and wild P. heterophylla . Due to the lack of a genomic database, we used high-throughput transcriptomic and proteomic technologies to identify proteins in the herb. Isobaric tags for relative and absolute quantification (iTRAQ) MS/MS were used to detect statistically significant changes between cultivated and wild P. heterophylla . We detected 3775 proteins; 332 showed differential accumulations across two different ecotypes of P. heterophylla . 71 significant differential expressions of proteins were selected based on GO annotations, KEGG, STRING analysis, and expression level. Quantitative real-time PCR analysis confirmed the authenticity and accuracy of the proteomic analysis. The results indicated that the carbohydrate and cellular amino acid metabolisms in cultivated P. heterophylla were weaker than those in its wild-type; seven important proteins were found to regulate sucrose and amino acids. This will provide the basic information for exploring the cause of differences in secondary metabolites in different ecotypes of P. heterophylla and the protein mechanism of its quality formation. Biological significance This study combined a transcriptome, proteome, and metabolism approach for analyzing differentially expressed proteins of cultivated and wild P. heterophylla and established the relationship between significantly differentially expressed proteins and differential chemical components in non-model plants. The results of proteomic analysis provide the basic information for exploring the quality forming process, which will demonstrate, and provide guidance for, the study of effective constituents of P. heterophylla and its quality formation mechanism. [ABSTRACT FROM AUTHOR]

• The first attempt to employ proteomics to elucidate salt tolerance mechanism of Apocyni Veneti Folium. • Key salt-responsive proteins, such as dehydrin 1, annexin, PRH, POP, POD, CAD, 4CL3, CYP were identified. • Carbohydrate and energy metabolism, primary and secondary metabolites and signal transduction contribute to salt tolerance. • Novel information for halophyte quality formation was provided. Soil salinity is a major abiotic stress that limits plant growth and productivity. Understanding the mechanisms of plant salinity tolerance can facilitate engineering for quality improvement. Apocynum venetum L. exhibits tolerance to salinity. Due to the lack of a genomic database, RNA-seq based transcriptomics and isobaric tag for relative and absolute quantitation (iTRAQ) based proteomic profiles of Apocyni Veneti Folium (leaves of Apocynum venetum L.) exposure to four levels of salt treatments (0, 100, 200 and 300 mM NaCl, respectively) were performed. A total of 143, 162 and 167 differentially expressed proteins (DEPs) were found between salt-treated Apocyni Veneti Folium compared with control, respectively. They were mainly involved in carbohydrate and energy metabolism, biosynthesis of metabolites and signal transduction. Furthermore, results showed that carbon and nitrogen metabolisms were altered under salt stress; low and moderate levels of salt stress enhanced photosynthetic functions and ramped up carbohydrate metabolism. However, severe salt stress depressed biosynthesis of secondary metabolites, consistent with the metabolomics results. It is worth emphasizing that some key salt-responsive proteins, such as dehydrin 1, annexin, pathogenesis-related protein, prolyl oligopeptidase, peroxidase, cinnamyl alcohol dehydrogenase, 4-hydroxycinnamoyl-CoA ligase 3, cytochrome P450 CYP73A120, were screened. These novel proteins provide a good starting point for further research into their functions using genetic or other approaches. In addition, a weak correlation between the abundance of DEPs and the corresponding differentially expressed genes highlighted the effect of post-transcriptional modifications and the importance of employing proteomics and transcriptomics to analyze global protein level changes. In conclusion, the protein profiles indicate that halophyte uses a multipronged approach to overcome salt stress, and provides some novel information on revealing the mechanisms of adaption and quality formation. [ABSTRACT FROM AUTHOR]