Impact of Methyl Jasmonate on Terpenoid Biosynthesis and Functional Analysis of Sesquiterpene Synthesis Genes in Schizonepeta tenuifolia.

This study investigates the impact of methyl jasmonate (MeJA) on the volatile oil composition of Schizonepeta tenuifolia and elucidates the function of the StTPS45 gene, a key player in terpenoid biosynthesis. The effect of different concentrations of MeJA (0, 50, 100, 200, and 300 µmol/L) on the growth of S. tenuifolia adventitious bud clusters was analyzed over a 20 d period. Using gas chromatography-mass spectrometry (GC-MS), 17 compounds were identified from the adventitious bud clusters of S. tenuifolia. Significant changes in the levels of major monoterpenes, including increased contents of (+)-limonene and (+)-menthone, were observed, particularly at higher concentrations of MeJA. Analysis of transcriptome data from three groups treated with 0, 100, and 300 µmol/L MeJA revealed significant changes in the gene expression profiles following MeJA treatment. At 100 µmol/L MeJA, most terpene synthase (TPS) genes were overexpressed. Additionally, gene expression and functional predictions suggested that StTPS45 acts as germacrene D synthase. Therefore, StTPS45 was cloned and expressed in Escherichia coli, and enzyme activity assays confirmed its function as a germacrene D synthase. Molecular docking and structural prediction of StTPS45 further suggested specific interactions with farnesyl diphosphate (FPP), aligning with its role in the terpenoid synthesis pathway. These findings provide valuable insights into the modulation of secondary metabolite pathways by jasmonate signaling and underscore the potential of genetic engineering approaches to enhance the production of specific terpenoids in medicinal plants.

Heat Stress and Microbial Stress Induced Defensive Phenol Accumulation in Medicinal Plant Sparganium stoloniferum.

An approach based on the heat stress and microbial stress model of the medicinal plant Sparganium stoloniferum was proposed to elucidate the regulation and mechanism of bioactive phenol accumulation. This method integrates LC-MS/MS analysis, 16S rRNA sequencing, RT-qPCR, and molecular assays to investigate the regulation of phenolic metabolite biosynthesis in S. stoloniferum rhizome (SL) under stress. Previous research has shown that the metabolites and genes involved in phenol biosynthesis correlate to the upregulation of genes involved in plant-pathogen interactions. High-temperature and the presence of Pseudomonas bacteria were observed alongside SL growth. Under conditions of heat stress or Pseudomonas bacteria stress, both the metabolites and genes involved in phenol biosynthesis were upregulated. The regulation of phenol content and phenol biosynthesis gene expression suggests that phenol-based chemical defense of SL is stimulated under stress. Furthermore, the rapid accumulation of phenolic substances relied on the consumption of amino acids. Three defensive proteins, namely Ss4CL, SsC4H, and SsF3'5'H, were identified and verified to elucidate phenol biosynthesis in SL. Overall, this study enhances our understanding of the phenol-based chemical defense of SL, indicating that bioactive phenol substances result from SL's responses to the environment and providing new insights for growing the high-phenol-content medicinal herb SL.

GC-MS Combined with Fast GC E-Nose for the Analysis of Volatile Components of Chamomile (Matricaria chamomilla L.).

Chamomile has become one of the world's most popular herbal teas due to its unique properties. Chamomile is widely used in dietary supplements, cosmetics, and herbal products. This study aimed to investigate the volatile aromatic components in chamomile. Two analytical techniques, gas chromatography-mass spectrometry (GC-MS) and an ultra-fast gas chromatography electronic nose, were employed to examine samples from Xinjiang (XJ), Shandong (SD), and Hebei (HB) in China, and imported samples from Germany (GER). The results revealed that all chamomile samples contained specific sesquiterpene compounds, including α-bisabolol, bisabolol oxide, bisabolone oxide, and chamazulene. Additionally, forty potential aroma components were identified by the electronic nose. The primary odor components of chamomile were characterized by fruity and spicy notes. The primary differences in the components of chamomile oil were identified as (E)-ß-farnesene, chamazulene, α-bisabolol oxide B, spathulenol and α-bisabolone oxide A. Significant differences in aroma compounds included geosmin, butanoic acid, 2-butene, norfuraneol, γ-terpinene. This study demonstrates that GC-MS and the ultra-fast gas chromatography electronic nose can preliminarily distinguish chamomile from different areas, providing a method and guidance for the selection of origin and sensory evaluation of chamomile. The current study is limited by the sample size and it provides preliminary conclusions. Future studies with a larger sample size are warranted to further improve these findings.

Effect of different fertilization strategies on the yield, quality of Euryales Semen and soil microbial community.

Introduction: Euryales Semen, a medicinal herb widely utilized in Asia, faces a critical constraint in its production, primarily attributed to fertilizer utilization. Understanding the impact of different fertilization schemes on Euryales Semen (ES) planting and exploring the supporting mechanism are crucial for achieving high yield and sustainable development of the ES planting industry. Methods: In this study, a field plot experiment was conducted to evaluate the effects of four different fertilization treatments on the yield and quality of ES using morphological characteristics and metabolomic changes. These treatments included a control group and three groups with different organic fertilizer to chemical fertilizer ratios (3:7, 5:5, and 7:3). The results of this study revealed the mechanisms underlying the effect of the different treatments on the yield and quality of Euryales Semen. These insights were achieved through analyses of soil physicochemical properties, soil enzyme activity, and soil microbial structure. Results: We found that the quality and yield of ES were the best at a ratio of organic fertilizer to chemical fertilizer of 7:3. The optimality of this treatment was reflected in the yield, soil available nitrogen, soil available phosphorus, and soil enzyme activity of ES. This ratio also increased soil microbial diversity, resulting in an increase and decrease in Proteobacteria and Firmicutes abundances, respectively. In addition, linear discriminant analysis showed that Chloroflexi, Gammaproteobacteria, and Hypocreales-incertae-sedis were significantly enriched in the ratio of organic fertilizer to chemical fertilizer of 7:3. Variance partitioning analysis showed that the soil properties, enzyme activities, and their interactions cumulatively can explain 90.80% of the differences in Euryales Semen yield and metabolome. In general, blending organic and chemical fertilizers at a 7:3 ratio can enhance soil fertility, boost Euryales Semen yield and quality, and bring forth conditions that are agriculturally beneficial to microbial (bacteria and fungi) dynamics. Discussion: This study initially revealed the scientific connotation of the effects of different fertilization patterns on the planting of Euryales Semen and laid a theoretical foundation for the study of green planting patterns of Euryales Semen with high quality and yield.

Metabolite profiling and transcriptomic analyses demonstrate the effects of biocontrol agents on alkaloid accumulation in Fritillaria thunbergii.

BACKGROUND: During Fritillaria thunbergii planting, pests and diseases usually invade the plant, resulting in reduced yield and quality. Previous studies have demonstrated that using biocontrol agents can effectively control grubs and affect the steroid alkaloids content in F. thunbergii. However, the molecular regulatory mechanisms underlying the differences in the accumulation of steroid alkaloids in response to biocontrol agents remain unclear. RESULTS: Combined transcriptomic and metabolic analyses were performed by treating the bulbs of F. thunbergii treated with biocontrol agents during planting. Otherwise, 48 alkaloids including 32 steroid alkaloids, 6 indole alkaloids, 2 scopolamine-type alkaloids, 1 isoquinoline alkaloid, 1 furoquinoline alkaloid, and 6 other alkaloids were identified. The content of steroidal alkaloids particularly peimine, peiminine, and veratramine, increased significantly in the group treated with the biocontrol agents. Transcriptome sequencing identified 929 differential genes using biocontrol agents, including 589 upregulated and 340 downregulated genes. Putative biosynthesis networks of steroid alkaloids have been established and combined with differentially expressed structural unigenes, such as acetyl-CoA C-acetyl-transferase, acelyl-CoAC-acetyltransferase3-hydroxy-3-methylglutaryl-coenzyme A synthase, 1-deoxy-D-xylulose-5-phosphate reductor-isomerase, 2-C-methyl-D-erythritol-4-phosphate cytidylyltransferase and 4-hydroxy-3-methylbut-2-enyl diphosphate reductase. In addition, biological processes such as amino acid accumulation and oxidative phosphorylation were predicted to be related to the synthesis of steroid alkaloids. Cytochrome P450 enzymes also play crucial roles in the steroid alkaloid synthesis. The transcription factor families MYB and bHLH were significantly upregulated after using biocontrol agents. CONCLUSIONS: Biocontrol agents increased the steroid alkaloids accumulation of steroid alkaloids by affecting key enzymes in the steroid alkaloid synthesis pathway, biological processes of oxidative phosphorylation and amino acid synthesis, cytochrome P450 enzymes, and transcription factors. This study revealed the mechanism underlying the difference in steroidal alkaloids in F. thunbergii after using biocontrol agents, laying the groundwork for future industrial production of steroid alkaloids and ecological planting of medicinal materials in the future.

Chemical profiling and quantification analysis of flavonoids in different varieties of Euryales Semen by ultra-high-performance liquid chromatography with tandem mass spectrometry.

Euryales Semen was a traditional Chinese medicine, which has been commonly used to treat spermatorrhea, enuresis, and frequent urination. Flavonoids were a critical ingredient in determining the function and quality of Euryales Semen. At present, no effective method has been established for the qualitative of Euryales Semen flavonoids. In this study, an ultra-high-performance liquid chromatography-quadrupole-time of flight-mass spectrometry method was established for flavonoids. By comparison with standard or literature data, 32 flavonoid compounds have been identified in Euryales Semen. Based on the qualitative results, an ultra-high-performance liquid chromatography-triple quadrupole tandem mass spectroscopy method was developed for the main components, and the linearity, the limit of detection, limit of quantification, repeatability, precision, stability, and recovery of the method were verified. The principal component analysis and the hierarchical clustering heatmaps analysis showed that the 30 batches of samples were distinctly separated into the North Gordon Euryale and South Gordon Euryale, and the measured contents of the six flavonoids in North Gordon Euryale were more abundant than in South Gordon Euryale, especially isoquercitrin, hesperetin, and quercetin. It provided a scientific basis for the quality control of Euryales Semen and a theoretical basis for the rational utilization of Euryales Semen resources.