Comparative transcriptome analysis and HPLC reveal candidate genes associated with synthesis of bioactive constituents in Rheum palmatum complex.

Content of bioactive constituents is one of the most important characteristics in Rheum palmatum complex. Increasing ingredient content through genetic breeding is an effective strategy to solve the contradiction between large market demand and resource depletion, but currently hampered by limited understanding of metabolite biosynthesis in rhubarb. In this study, deep transcriptome sequencing was performed to compare roots, stems, and leaves of two Rheum species (PL and ZK) that show different levels of anthraquinone contents. Approximately 0.52 billion clean reads were assembled into 58,782 unigenes, of which around 80% (46,550) were found to be functionally annotated in public databases. Expression patterns of differential unigenes between PL and ZK were thoroughly investigated in different tissues. This led to the identification of various differentially expressed genes (DEGs) involved in shikimate, MEP, MVA, and polyketide pathways, as well as those involved in catechin and gallic acid biosynthesis. Some structural enzyme genes were shown to be significantly up-regulated in roots of ZK with high anthraquinone content, implying potential central roles in anthraquinone synthesis. Taken together, our study provides insights for future functional studies to unravel the mechanisms underlying metabolite biosynthesis in rhubarb. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-024-01492-z.

Study on the quality difference between raw and ginger juice processed Magnoliae officinalis cortex by UPLC-Q-TOF-MS/MS and GC-MS coupled with color measurement.

INTRODUCTION: Magnoliae officinalis cortex (MOC) has been used for thousands of years as a traditional Chinese herb. In Chinese Pharmacopoeia (2020 edition), it has two types of decoction pieces, raw Magnoliae officinalis cortex (RMOC) and ginger juice processed Magnoliae officinalis cortex (GMOC). The quality difference between RMOC and GMOC has not been explored systemically. OBJECTIVE: This study aimed to discover the quality difference between RMOC and GMOC, and clarify the effect of ginger juice during processing comprehensively. METHODS: Ultra-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF-MS/MS) and gas chromatography-mass spectrometry (GC-MS) were applied to study the non-volatile and volatile components of RMOC and GMOC; electronic eye was applied for color measurement. Meanwhile, water processed Magnoliae officinalis cortex (WMOC) was studied as the blank sample. RESULTS: There were 155 non-volatile and 72 volatile substances identified. Between RMOC and GMOC, 29 distinctive non-volatile and 34 distinctive volatile compounds were detected, among which 23 new compounds appeared and five compounds disappeared due to the addition of ginger juice during processing. The intensities of 12 common non-volatile compounds and the relative percentage contents of four common volatile compounds showed significant differences between RMOC and GMOC. In color measurement of RMOC, GMOC, and WMOC, 14 common compounds with significant differences were discovered related to their color values, and their mathematical prediction functions were built. CONCLUSION: There were significant differences between RMOC and GMOC; the processing mechanism of GMOC would be carried out based on the differential compounds in further investigation.

Amino acids and flavonoids analysis reveals quality constituents difference among different albino tea resources.

Albino tea has attracted increased attention due to its unique flavor. To reveal the difference in key metabolites constituting the important quality of different tea resources, amino acids and flavonoids profiles in three albino resources with different degrees of albinism and one normal green variety were comprehensively investigated. K-means analysis revealed 35 amino acids were significantly enriched in 'Jibai', while 3 and 2 were specifically accumulated in 'Huangjinya' and 'Anjibaicha', respectively. Based on OPLS-DA models, 40, 31 and 45 significantly differential flavonoids were determined in 'Huangjinya', 'Anjibaicha' and 'Jibai' compared to 'Fudingdabaicha', and most were down-regulated. Among them, 10, 5 and 13 differential flavonoids were exclusively found in 'Huangjinya', 'Anjibaicha' and 'Jibai', respectively, which may contribute to unique quality for different resources. The differential flavonoids and amino acids involved in their metabolic pathways were obviously different among four resources, resulting in the difference in tea quality and flavor.

An evaluation of Astragali Radix with different growth patterns and years, based on a new multidimensional comparison method.

Introduction: With the depletion of wild Astragali Radix (WA) resources, imitated-wild Astragali Radix (IWA) and cultivated Astragali Radix (CA) have become the main products of Astragali Radix. However, the quality differences of three growth patterns (WA, IWA, CA) and different growth years of Astragali Radix have not been fully characterized, leading to a lack of necessary scientific evidence for their use as substitutes for WA. Methods: We innovatively proposed a multidimensional evaluation method that encompassed traits, microstructure, cell wall components, saccharides, and pharmacodynamic compounds, to comprehensively explain the quality variances among different growth patterns and years of Astragali Radix. Results and discussion: Our study showed that the quality of IWA and WA was comparatively similar, including evaluation indicators such as apparent color, sectional structure and odor, thickness of phellem, diameter and number of vessels, morphology of phloem and xylem, and the levels and ratios of cellulose, hemicellulose, lignin, sucrose, starch, water-soluble polysaccharides, total-saponins. However, the content of sucrose, starch and sorbose in CA was significantly higher than WA, and the diameter and number of vessels, total-flavonoids content were lower than WA, indicating significant quality differences between CA and WA. Hence, we suggest that IWA should be used as a substitute for WA instead of CA. As for the planting years of IWA, our results indicated that IWA aged 1-32 years could be divided into three stages according to their quality change: rapid growth period (1-5 years), stable growth period (6-20 years), and elderly growth period (25-32 years). Among these, 6-20 years old IWA exhibited consistent multidimensional comparative results, showcasing elevated levels of key active components such as water-soluble polysaccharides, flavonoids, and saponins. Considering both the quality and cultivation expenses of IWA, we recommend a cultivation duration of 6-8 years for growers. In conclusion, we established a novel multidimensional evaluation method to systematically characterize the quality of Astragali Radix, and provided a new scientific perspective for the artificial cultivation and quality assurance of Astragali Radix.

Exploring major variable factors influencing flavor and microbial characteristics of upper jiupei.

The flavor quality of jiupei gradually decreased with the increase of cellar height. In this study, high-throughput sequencing, metabolomics and HS-SPME-GC-MS techniques were used to explore the mechanism of flavor quality decline in upper jiupei in mud sealed cellars. The results showed the total content of flavor compounds increased from 1947.48 mg/L in top-site to 3855.51 mg/L in bottom of the cellar, and 19 differential flavor compounds were identified based on OPLS-DA, mainly including 12 esters such as ethyl hexanoate, ethyl butyrate, propyl hexanoate, hexyl caproate and 5 other substances such as caprylic acid, decanal and nonaldehyde. Lactobacillus, Prevotella and Methanobacterium were dominant genus of bacteria in all of cellars, while Thermomyces, Aspergillus, Pichia, Trichosporon and Rhizopus were the dominant genera of fungi. Oxygen was the key factor causing the quality heterogeneity of flavor substances and microbial communities in jiupei at different depths. Anaerobic micro-pressure sealed cellars (AMSC) method was developed and applied to jiupei fermentation, the difference in oxygen content between top site (5.90 ± 0.62 %) and bottom of the cellar (4.17 ± 0.75 %) in AMSC was smaller than that in mud sealed cellars, there were no significant differences in flavor substances content between top site and bottom of the cellar, and microbial communities showed no significant differences of the four-layer jiupei. This study provides a theoretical support for improving the flavor quality of upper jiupei.

Revealing ecotype influences on Cistanche sinensis: from the perspective of endophytes to metabolites characteristics.

Introduction: Plant microorganism is critical to plant health, adaptability, and productive forces. Intriguingly, the metabolites and microorganisms can act upon each other in a plant. The union of metabolomics and microbiome may uncover the crucial connections of the plant to its microbiome. It has important benefits for the agricultural industry and human being health, particularly for Chinese medical science investigation. Methods: In this last 2 years study, on the strength of the UPLC-MS/MS detection platform, we accurately qualitatively, and quantitatively measured the Cistanche sinensis fleshy stems of two ecotypes. Thereafter, through high-throughput amplicon sequencing 16S/ITS sequences were procured. Results: PhGs metabolites including echinacoside, isoacteoside, and cistanoside A were significantly downregulated at two ecotypes of C. sinensis. Add up to 876 metabolites were monitored and 231 differential metabolites were analyzed. Further analysis of 34 core differential metabolites showed that 15 compounds with up-regulated belonged to phenolic acids, flavonoids, and organic acids, while 19 compounds with down-regulated belonged to phenolic acids, flavonoids, alkaloids, amino acids, lipids, and nucleotides. There was no noteworthy discrepancy in the endophytic bacteria's α and ß diversity between sandy and loam ecotypes. By comparison, the α and ß diversity of endophytic fungi was notably distinct. The fungal community of the loam ecotype is more abundant than the sandy ecotype. However, there were few such differences in bacteria. Most abundant genera included typical endophytes such as Phyllobacterium, Mycobacterium, Cistanche, Geosmithia, and Fusarium. LEfSe results revealed there were 11 and 20 biomarkers of endophytic bacteria and fungi in C. sinensis at two ecotypes, respectively. The combination parsing of microflora and metabolites indicated noteworthy relativity between the endophytic fungal communities and metabolite output. Key correlation results that Anseongella was positive relation with Syringin, Arsenicitalea is negative relation with 7-methylxanthine and Pseudogymnoascus is completely positively correlated with nepetin-7-O-alloside. Discussion: The aim of this research is: (1) to explore firstly the influence of ecotype on C. sinensis from the perspective of endophytes and metabolites; (2) to investigate the relationship between endophytes and metabolites. This discovery advances our understanding of the interaction between endophytes and plants and provides a theoretical basis for cultivation of C. sinensis in future.

Widely Targeted Metabolomics Profiling Reveals the Effect of Powdery Mildew on Wine Grape Varieties with Different Levels of Tolerance to the Disease.

Powdery mildew is an economic threat for viticulture because it not only affects grape yield, but also causes a series of impacts on the qualities of fruit and wine, especially the flavors and various metabolites. Different grape varieties may have different levels of powdery mildew resistance/tolerance and their components of their metabolome are also various. In this study, two wine grape varieties, Guipu No.6 (GP6) and Marselan (Mar) with different levels of powdery mildew tolerance, were used to compare the quality differences in metabolism level by using the widely targeted metabolomics method. The results show that GP6 has a better powdery mildew leaf tolerance than Mar. A total of 774 metabolites were detected by using a UPLC-QQQ-MS-based metabolomics approach, and 57 differential metabolites were identified as key metabolites that were accumulated after infection with powdery mildew in GP6 and Mar, including phenolic acids, flavonoids, terpenoids, stilbenes, lipids, nucleotides and derivatives, lignans and coumarins, and quinones. This finding indicates that the defense mechanisms of grape fruit are mainly associated with phenylpropane-flavonoid metabolism. Specifically, stilbenes had greater variations after powdery mildew infection in GP6; while in Mar, the variations of flavonoids, especially kaempferol-3-O-glucuronide and luteolin-7-O-glucuronide, were more remarkable. The above results demonstrate that stilbenes may play a more important role than flavonoids in resisting powdery mildew infection in GP6's fruits, and the drastic variations of these phenolic compounds in different wine grapes after powdery mildew infection might also lead to quality difference in the flavors. This study can provide new insights into the understanding of the cause of powdery mildew tolerance in different grape varieties and the effects on the quality of wine grapes infected with the disease exerted by metabolism level.

Quality Difference Characteristics of Bupleurum Formulae Granules Based on the Analysis of Dual-Wavelength Fingerprint Profiles and Chemometric Methods / 中国药学杂志

OBJECTIVE: To distinguishing the Bupleurum formulae granules from different origins and promoting the consistency of quality of them,with the analysis of fingerprint profiles and chemometric methods. METHODS: In the study, a method of dual-wavelength (210 & 254 nm) HPLC fingerprint of Bupleurum formulae granules was developed. Based on the fingerprint method, the quality differences of the samples of Bupleurum formulae granules were analyzed by similarity analysis, hierarchical cluster analysis, principal component analysis and partial least squares discriminant analysis. RESULTS: The fingerprints of 12 batchs of Bupleurum formulae granules are established. The results indicated that the fingerprints of Bupleurum formulae granules from different origins could be differentiated well and the characteristic peaks could be marked. CONCLUSION: The 210 nm fingerprints of Bupleurum formulae granule have the most characteristic differences; It is recommended that the dual-wavelength fingerprints could be combined for analysis and application. Saikosaponin a, saikosaponin b2 and saikosaponin b1 are the key components of the quality difference, which should be focused on in the quality evaluation and control of Bupleurum formula granules.