Amomum Tsao-ko Research Articles

Complete genome sequence of tsaoko stripe mosaic virus, a novel macluravirus found in Amomum tsaoko.

A novel macluravirus, tentatively named "tsaoko stripe mosaic virus" (TkSMV), was identified in Amomum tsaoko through high-throughput sequencing. The complete genome sequence of TkSMV was determined using RT-PCR and RACE. The genome sequence consists of 8218 nucleotides, excluding the poly(A) tail, and contains a large open reading frame encoding a polyprotein of 2625 amino acids with a molecular weight of approximately 297.13 kDa. TkSMV is most closely related to Alpinia oxyphylla mosaic virus, sharing 71.5% nucleotide and 75.9% amino acid sequence identity. These values are below the species demarcation threshold for the family Potyviridae. These results suggest that TkSMV should be considered a distinct member of the genus Macluravirus.

Isospongian Diterpenoids from the Leaves of Amomum tsao-ko Promote GLP-1 Secretion via Ca2+/CaMKII and PKA Pathways and Inhibit DPP-4 Enzyme.

Three uncommon isospongian diterpenoids including a new one, 3-epi-kravanhin A (2), were isolated from the leaves of Amomum tsao-ko. Compounds 2 and 3 dose-dependently promoted GLP-1 secretion on STC-1 cells with promotion ratios of 109.7% and 186.1% (60 μM). Mechanism study demonstrated that the GLP-1 stimulative effects of 2 and 3 were closely related with Ca2+/CaMKII and PKA pathways, but irrelevant to GPBAR1 and GPR119 receptors. Moreover, compound 1 showed moderate DPP-4 inhibitory activity with an IC50 value of 311.0 μM. Molecular docking verified the binding affinity of 1 with DPP-4 by hydrogen bonds between the γ-lactone carbonyl (C-15) and Arg61 residue. Bioinformatics study indicated that compound 1 exerted antidiabetic effects by improving inflammation, oxidative stress and insulin resistance. This study first disclosed the presence of isospongian diterpenoids in A. tsao-ko, which showed antidiabetic potency by promoting GLP-1 secretion and inhibiting DPP-4 activity.

Analysis of Chemical Changes during Maturation of Amomum tsao-ko Based on GC-MS, FT-NIR, and FT-MIR.

Amomum tsao-ko Crevost et Lemaire (A. tsao-ko) is widely grown for its high nutritional and economic value. However, the lack of a scientific harvesting and quality control system has resulted in an uneven product quality. The present study was based on A. tsao-ko from four maturity stages from the same growing area, and its chemical trends and quality were evaluated using a combination of agronomic trait analysis, spectroscopy, chromatography, chemometrics, and network pharmacology. The results showed that A. tsao-ko was phenotypically dominant in October. Spectroscopy showed that the absorbance intensity at different maturity stages showed a trend of October > September > August > July. Further chemical differences between A. tsao-ko at different stages of maturity were found by chromatography to originate mainly from alcohol, aromatic, acids, esters, hydrocarbons, ketone, heterocyclic, and aldehydes. The network pharmacology results showed that the active ingredient for the treatment of obesity was present in A. tsao-ko and had high levels in A. tsao-ko in September and October. The results of this study provide a new idea for the comprehensive evaluation of A. tsao-ko and a theoretical basis for the harvesting and resource utilization of A. tsao-ko.

Analysis of the Volatile Components in Different Parts of Three Species of the Genus Amomum via Combined HS-SPME-GC-TOF-MS and Multivariate Statistical Analysis.

The study used headspace solid-phase microextraction coupled with gas chromatography-time-of-flight mass spectrometry (HS-SPME-GC-TOF-MS) to analyze volatile compounds in leaves and fruits of Amomum tsaoko, Amomum paratsaoko, and Amomum koenigii. The composition and aroma of distinct metabolites were analyzed using multivariate statistical methods. A total of 564 volatile compounds were identified from three species of the genus Amomum, which were further divided into nine categories: terpenoids, carboxylic acids, alcohols, hydrocarbons, aldehydes, ketones, phenols, ethers, and other compounds. Terpenoids and alcohols were the most abundant. The content and types of compounds vary in A. tsaoko, A. paratsaoko, and A. koenigii, so mixing or substituting them is not advisable. We selected 45 metabolites based on the criteria of the variable importance in projection values (VIP > 1.5) and one-way ANOVA (p < 0.05). The top 19 metabolites with the most significant VIP values were chosen. Interestingly, (Z)-2-decenal was only found in Amomum koenigii, while nitroethane and nonanal were only present in cultivated A. tsaoko. Additionally, linalool, cineole, and (D)-limonene were the main components affecting the aroma of three species of the genus Amomum. The volatile components identified in this study provide a theoretical basis for analyzing the unique flavor of A. tsaoko, A. paratsaoko, and A. koenigii.

Antibacterial and gut health effects of Amomum tsao-ko in aquatic feed: A sustainable alternative to chemical antibiotics.

Amomum tsao-ko Crevost et Lemarie (Zingiberaceae), an aromatic plant, has been considered to have diverse medicinal values and economic significance. It has been reported to possess antibacterial, antioxidant, and antidiabetic effects. With the increasing risk of diseases in aquaculture, there is a need for alternative solutions to chemical antibiotics. Plant extracts have shown promise as natural feed additives for aquatic animals. In this study, the antibacterial effect of Amomum tsao-ko crude extracts was evaluated using the Oxford cup method. The extracts exhibited significant antimicrobial activity against Salmonella typhimurium and Salmonella enteritidis. Furthermore, the addition of Amomum tsao-ko to fish feed resulted in notable changes in the gut structure of zebrafish and tilapia. The length and morphology of intestinal villi were enhanced, promoting improved digestion. Analysis of the gut microbial community revealed that Amomum tsao-ko supplementation induced key changes in the gut microbial community composition of both zebrafish and tilapia. Notably, a 1% inclusion of Amomum tsao-ko resulted in a marked rise in Proteobacteria levels in zebrafish, which diminished at 10% dosage. The supplement elicited mixed reactions among other bacterial phyla like Actinobacteria and Verrucomicrobiota. Fluctuations were also observed at the genus level, pointing to the concentration of Amomum tsao-ko playing a pivotal role in influencing the structure of intestinal bacteria. The findings of this study suggest that Amomum tsao-ko has antibacterial properties and can positively influence the gut health of fish. The potential use of Amomum tsao-ko as a natural feed additive holds promise for improving aquaculture practices and reducing reliance on chemical antibiotics. Further research is needed to explore the full potential and applications of Amomum tsao-ko in fish feed development.

Comparison of the effects of Amomum tsaoko and its adulterants on functional dyspepsia rats based on metabolomics analysis

Amomum tsaoko (AT) is commonly used in clinical practice to treat abdominal distension and pain. It is also a seasoning for cooking, with the functions of appetizing, invigorating the spleen, and being digestive-promoting. Amomum tsaoko (AT) has three adulterants, Amomum paratsaoko (AP), Amomum koenigii (AK), and Alpinia katsumadai Hayata, because of the confusion in historical classics regarding recorded sources as well as the near geographic distribution and fruit morphological similarities. In this study, we established a functional dyspepsia (FD) rat model and then treated it with the corresponding medicinal solutions AT, AP, AK, and AKH. The gastric emptying rate, intestinal propulsion rate, serum biochemical indicators, histopathological changes, and fecal metabolism were measured. The efficacy and mechanism of AT, AP, AK, and AKH in the treatment of FD were compared. Fecal metabolomics revealed that 20 potential biomarkers were involved in seven significant metabolic pathways in FD rats. These pathways include ubiquinone and other terpenoid-quinone biosynthesis, glycerophospholipid metabolism, tyrosine metabolism, primary bile acid biosynthesis, purine metabolism, folate biosynthesis, and amino sugar and nucleotide sugar metabolism. AP regulates 6 metabolic pathways, 5 metabolic pathways affected by AT, 4 metabolic pathways affected by AK, and 2 metabolic pathways affected by AKH.The above results suggest that the different effects of AT, AP, AK, and AKH on FD rats may be due to their different regulatory effects on the metabolome.

Insights into the key odorants in fresh and dried Amomum tsaoko using the sensomics approach

To identify the key odorants in Amomum tsaoko (AT), volatiles in fresh AT (FAT) and dried AT (DAT) were investigated using molecular sensory science. In addition to this, the sensomics approach was used to confirm the presence of the compound in FAT that contributed the most to its aroma profile. A total of 49 odor-active compounds (43 in FAT and 42 in DAT) with flavor dilution (FD) factors ranging from 1 to 6561 were identified, with eucalyptol exhibiting the highest FD factor of 6561. Odorants with FD factors ≥ 27 were quantitated, and 23 and 20 compounds in FAT and DAT, respectively, with odor activity value ≥ 1 were determined as key odorants. Recombination and omission experiment further indicated that (E)-2-dodecenal, geranial, octanal, (E)-2-octenal, (E)-2-decenal, and eucalyptol contributed significantly to the overall aroma profile of FAT. After drying of FAT, the concentrations of aldehydes decreased significantly, whereas those of terpene hydrocarbons increased. Multivariate statistical analysis revealed that 26 FAT and 23 DAT odorants were biomarker compounds.

Differences between two plants fruits: Amomum tsaoko and Amomum maximum, using the SPME-GC–MS and FT-NIR to classification

Also known as Amomum tsaoko Crevost et Lemaire (A. tsaoko), is a spice and medicinal fruit plant that can be used in daily human life for cooking and health purposes. In Xishuangbanna (Yunnan Province, China) there is an interesting phenomenon where the Amomum maximum Roxb. (A. maximum) is consumed as a fruit by the local people. The spice trade played a significant role in the development of human civilization and the histories of nations. Both of them have high edible and medicinal value. The identification and classification of the metabolic components of both are key to driving the development of related industries. We used SPME-GC–MS (Non-targeted headspace-solid phase microextraction-gas chromatography-mass spectrometry) for its accurate qualitative and quantitative performance, combined with multiple analytical methods, to explain the metabolic components present in the fruits of these two plants. Next, near-infrared (NIR) technology offers researchers the advantage of saving time and being cost-effective. We analyzed the characteristic spectral chemical information of the NIR data. Identified the relevant base group and vibration mode. After analyzing the data of both, it was determined that there are a total of 66 metabolites, with 64 from cardamom and 29 from the substance, showing 37 differences. Among the metabolites, 35 had a significance level of P ≤ 0.05 and VIP ≥ 1, totaling 101 substances. This includes terpenes (37.9 %), aldehydes (12.1 %), organic heterocyclic compounds (7.6 %), alcohols (6.1 %), aromatic compounds (4.5 %), esters (4.5 %), and ketones (4.5 %). This study validates the metabolic components of the two species and provides a clear classification, which will be beneficial for future data-based applications in the fields of food and medicine.

Screening, evaluation and identification of promising plant extracts for development of novel natural preservatives

Chemical preservatives are widely employed as the primary approach to inhibit food spoilage, yet their acceptance among consumers is progressively diminishing. Consequently, researchers have increasingly made efforts to explore natural preservatives. Phytochemicals with diverse bioactivities offer promising opportunities for development of natural preservatives. In this study, extracts from a total of 90 plant materials were screened and evaluated for their inhibitory effects against the two pathogenic fungi, Aspergillus flavus and Penicillium citrinum. Initially, 20 plant extract samples that exhibited an inhibition rate exceeding 85 % against either or both fungal strains were conducted for further screening. Then, evaluations of their minimum inhibition concentration (MIC), spore germination, and mycelial growth were carried out for the subsequent screenings. Finally, GC-MS analysis was preformed to detect the chemical composition of the selected plant extracts. Thus, the antifungal index of the 90 plants was calculated, forming a repository of raw material for plant-based preservatives. The results revealed significant antifungal activities in the petroleum ether (PE) fractions of Cinnamomum cassia, Syzygium aromaticum, Amomum tsaoko, Nardostachys jatamansi, Angelica sinensis, and chloroform fractions of Cinnamomum cassia, Syzygium aromaticum, Houpoea officinalis and Ruta graveolens. Especially, the MICs of Cinnamomum cassia PE fraction (CMPE) against the Aspergillus flavus and Penicillium citrinum were only 0.125 mg/mL and 0.25 mg/mL, respectively. Moreover, CMPE also exhibited strong inhibiton effects on sprore germination (> 90 %) and mycelial growth (> 50 %, 1 day). These plant extract fractions are considered as potential candidates for novel natural preservatives.

Tsaokoflavanols A1–J1: Flavanol-fatty alcohol hybrids with HPL inhibitory activity from Amomum tsao-ko

Ten previously undescribed compounds were isolated from the fruits of Amomum tsao-ko (Zingiberaceae), including nine undescribed flavanol-fatty alcohol hybrids (1–6, 10−11, 13), and a flavanol-monoterpenoid hybrid (14), along with seven known flavanol hybrids (7−9, 12, 15−17). The structures of these compounds were determined using various analyses, such as HRESIMS, 1D/2D NMR, and ECD calculations. In terms of biological activity, compounds 1, 2, 5, and 6 exhibited inhibitions of human pancreatic lipase (HPL), with IC50 values ranging from 0.017 to 0.193 mM. Some of these values were found to be stronger than that of the positive control, orlistat (IC50, 0.067 mM). Molecular docking studies were also conducted to investigate the interactions between these compounds and HPL. The docking simulations revealed the importance of the orientation of the 3,4-dihydroxyphenyl in binding with HPL. Additionally, compound 9 demonstrated cytotoxicity against HepG2, with a CC50 value of 14.96 ± 0.62 μM as determined by the MTT assay. Flow cytometry analysis indicated that compound 9 induced apoptosis in HepG2 cells. Western blot results showed an up-regulation of apoptosis-related proteins, such as p53 protein, Bax and Caspase-3 proteins, while the expression of Bcl-2 protein was down-regulated.

Antibacterial activity of Amomum tsaoko essential oil and its interaction with Staphylococcus aureus

Amomum tsaoko is a medicinal and edible plant mainly distributed in southwest China and Southeast Asia. This study investigated the antibacterial activity of Amomum tsaoko essential oil (AEO) against typical foodborne pathogenic bacteria and the interaction of AEO with Staphylococcus aureus. The results showed that AEO exhibited excellent antibacterial activity against foodborne pathogenic bacteria, with the most performance against S. aureus, with MIC and MBC values of 0.20 mg/mL and 0.39 mg/mL, respectively. In addition, the effects of AEO on S. aureus might include affecting cell membrane permeability, integrity, bacterial energy, and respiratory metabolism. Furthermore, the (E)-dec-2-enal in AEO was considered a vital antibacterial component. Overall, AEO presented positive antibacterial properties, suggesting that AEO can be served as a potential inhibitor against S. aureus and contribute to the application of AEO in food preservation.

An integrated chemical characterization based on FT-NIR, and GC–MS for the comparative metabolite profiling of 3 species of the genus Amomum

BackgroundThe fruits and seeds of genus Amomum are well-known as medicinal plants and edible spices, and are used in countries such as China, India and Vietnam to treat malaria, gastrointestinal disorders and indigestion. The morphological differences between different species are relatively small, and technical characterization and identification techniques are needed. ResultsFourier transform near infrared spectroscopy (FT-NIR) and gas chromatography-mass spectrometry (GC-MS), combined with principal component analysis and two-dimensional correlation analysis were used to characterize the chemical differences of Amomum tsao-ko, Amomum koenigii, and Amomum paratsaoko. The targets and pathways for the treatment of diabetes mellitus in three species were predicted using network pharmacology and screened for the corresponding pharmacodynamic components as potential quality markers. The results of “component-target-pathway” network showed that (+)-Nerolidol, 2-Nonanol, α-Terpineol, α-Pinene, 2-Nonanone had high degree values and may be the main active components. Partial least squares-discriminant analysis (PLS-DA) was further used to select for differential metabolites and was identified as a potential quality marker, 11 in total. PLS-DA and residual network (ResNet) classification models were developed for the identification of 3 species of the genus Amomum, ResNet model is more suitable for the identification study of large volume samples. SignificanceThis study characterizes the differences between the three species in a visual way and also provides a reliable technique for their identification, while demonstrating the ability of FT-NIR spectroscopy for fast, easy and accurate species identification. The results of this study lay the foundation for quality evaluation studies of genus Amomum and provide new ideas for the development of new drugs for the treatment of diabetes mellitus.

Investigation on the changes of odor-active compounds of Amomum tsao-ko during natural drying

To characterize the key odorants in Amomum tsao-ko (AT) and to investigate their changes during natural drying, volatile compounds of AT were extracted and analyzed by solvent extraction combined with solvent-assisted flavor evaporation and gas chromatographymass spectrometry–olfactometry. A total of 41 aroma-active compounds with flavor dilution factors ranging from 3 to 6561 were identified in AT. Fourteen odorants were determined to be the key odorants by quantitation analyses, odor activity value (OAV) calculations, recombination and omission experiments. To further investigate the changes in aroma components in AT during natural drying, the concentrations and OAVs of odor-active compounds were measured at 0, 2, 4, 6, 8, 12, 16, and 20 days. The results showed that the number of aroma-active components did not change during natural drying, but their content changed with different trends. The concentration of geraniol showed a continuous upward trend; those of hexanal, octanal, nonanal and pinocarvone decreased first and then increased; those of α-pinene, sabinene, 2-nonanone, (E)-2-octenal, 2-nonanol, 4-terpineol, epidolichodial and (E,E)-2,4-dodecadienal changed on a fluctuating trend, while those of the other 28 compounds showed a trend of first increasing and then decreasing. These results provide a theoretical basis for further utilization of AT.

Chemical markers of a rare honey from the traditional spice plant Amomum tsao–ko Crevost et Lemarié, via integrated GC–MS and LC-MS approaches

The precious medicinal plant, Amomum tsao-ko Crevost et Lemarié, is the nectariferous plant from which the rare Amomum tsao-ko Crevost et Lemarié honey (ATH) is produced. Presently, chemical markers for authentication of this honey are not available due to the lack of data on its chemical composition. Here, we analyzed the volatile components and their odor activity values (OAVs), which revealed that the unique aroma was mildly flowery and fruity, accompanied by subtle sweet and fresh undertones. Since non-volatile chemicals are more reliable markers for routine authentication, we used a metabolomic approach combined with NMR-based identification to find and confirm a suitable compound to unambiguously distinguish ATH from other honeys. Isorhamnetin 3-O-neohesperidoside ranged from 3.62 to 9.38 mg/kg in ATH and was absent in the other tested honeys. In sum, the study uncovered unique chemical characteristics of ATH that will be helpful to control its quality.

Purification, Characterization, and Anticomplementary Activity of Polysaccharides From the Fruits of Amomum tsaoko Crevost &amp; Lemarié

The resin purification condition of polysaccharides from Amomum tsaoko Crevost et Lemarie (ATPs) was optimized using response surface method (RSM) and analytic hierarchy process (AHP): elution volume of 2 BV, flow rate of 2 BV/h, sample concentration of 2 mg/mL, sample volume of 0.4 BV, the comprehensive score of actual purification effect was 73.45 ± 0.32% (n = 3). Then, a homogenous polysaccharide (ATP-1-1) was isolated by DEAE-52 cellulose and Sephadex G-200 column. The structure of ATP-1-1 was characterized by the Fourier transform infrared spectroscopy, high-performance liquid chromatography, high-performance gel permeation chromatography, Congo red test, scanning electron microscopy, and thermogravimetric analysis, which showed that ATP-1-1 was composed of mannose, rhamnose, glucose, galactose, and arabinose in a molar ratio of 1.3:1.0:6.2:4.8:1.1 with molecular weight of 845.21 kDa, and had a triple-helix conformation, blocky microstructure, and good thermal stability. The evaluation of anti-complementary activity showed that ATP-1-1 possessed stronger anticomplementary activity than positive drug, significantly inhibited complement activation through both classical (CH50 = 6.28 ± 0.23 μg/mL) and alternative pathways (AP50 = 45.33 ± 0.16 μg/mL), and interacted with C2, C4, and factor B components in the activation cascade of the complement system.

Widely Targeted Volatilomics and Metabolomics Analysis Reveal the Metabolic Composition and Diversity of Zingiberaceae Plants.

Zingiberaceae plants are widely used in the food and pharmaceutical industries; however, research on the chemical composition and interspecific differences in the metabolome and volatilome of Zingiberaceae plants is still limited. In this study, seven species of Zingiberaceae plants were selected, including Curcuma longa L., Zingiber officinale Rosc., Alpinia officinarum Hance, Alpinia tonkinensis Gagnep, Amomum tsaoko Crevost et Lemarie, Alpinia hainanensis K. Schum. and Amomum villosum Lour. Myristica fragrans Houtt. was also selected due to its flavor being similar to that of the Zingiberaceae plant. The metabolome and volatilome of selected plants were profiled by widely targeted approaches; 542 volatiles and 738 non-volatile metabolites were detected, and β-myrcene, α-phellandrene and α-cadinene were detected in all the selected plants, while chamigren, thymol, perilla, acetocinnamone and cis-α-bisabolene were exclusively detected in certain Zingiberaceae plants. Differential analysis showed that some terpenoids, such as cadalene, cadalene-1,3,5-triene, cadalene-1,3,8-triene and (E)-β-farnesene, and some lipids, including palmitic acid, linoleic acid and oleic acid were amongst the most varied compounds in Zingiberaceae plants. In conclusion, this study provided comprehensive metabolome and volatilome profiles for Zingiberaceae plants and revealed the metabolic differences between these plants. The results of this study could be used as a guide for the nutrition and flavor improvement of Zingiberaceae plants.

Amomum tsaoko DRM1 regulate seed germination and improve heat tolerance in Arabidopsis

Seed dormancy and germination are critical to medicinal plant reproduction. Dormancy-associated gene (DRM1) has been involved in the regulation of dormancy in Arabidopsis meristematic tissues or organs. However, research on molecular functions and regulations of DRM1 in Amomum tsaoko, an important medicinal plant, is rare. In this study, the DRM1 was isolated from embryos of A. tsaoko, and the results of protein subcellular localization in Arabidopsis protoplast indicated that DRM1 was mainly nucleus and cytoplasm. Expression analysis showed that DRM1 especially exhibited the highest transcript level in dormant seed and short-time stratification while displaying a high response of hormone and abiotic stress. Further investigation showed that ectopic expression of DRM1 in Arabidopsis exhibited delayed seed germination and germination capability to high temperatures. Additionally, DRM1 transgenic Arabidopsis exhibited increased tolerance to heat stress by enhancing antioxidative capacities and regulating stress-associated genes (AtHsp25.3-P, AtHsp18.2-CI, AtHsp70B, AtHsp101, AtGolS1, AtMBF1c, AtHsfA2, AtHsfB1 and AtHsfB2). Overall, our results reveal the role of DRM1 in seed germination and abiotic stress response.

A rapid identification based on FT-NIR spectroscopies and machine learning for drying temperatures of Amomum tsao-ko

Amomum tsao-ko (A. tsao-ko), is a medicine and food homology plant with high economic value. Different drying temperature affects flavor and quality of A. tsao-ko, which is closely related to its economic value and consumer acceptance. This research investigated the feasibility of combining Fourier-transform near infrared spectroscopy (FT-NIR) with chemometrics and machine learning to the identification of drying temperatures of A. tsao-ko. A total of 626 fresh samples from Honghe Prefecture, Yunnan, were dried to constant weight at 40 ℃ (62 h), 50 ℃ (54 h), 60 ℃ (43 h), 70 ℃ (35 h) and 80 ℃ (25 h) using an electric thermostatic drying oven. Principal component analysis (PCA) was used to observe the clustering of samples treated at different drying temperatures. Partial least squares-discriminant analysis (PLS-DA), support vector machines (SVM), and Residual Neural Network (ResNet) are used to build discriminant models. The accuracy of GS-SVM, GA-SVM, and ResNet models was more than 99% in the test set for identifying the drying temperature of A. tsao-ko. In contrast, the ResNet model based on synchronous two-dimensional correlation spectroscopy (2DCOS) images obtained the most satisfactory discrimination results. The use of appropriate data pre-processing (SD) can optimize the performance of the model to a certain extent and improve the accuracy of discrimination. This study demonstrates the feasibility of FT-NIR in the identification of the drying temperature of A. tsao-ko, providing a rapid and nondestructive method for the assessment and control of A. tsao-ko quality.

Metabolomics and Transcriptomics Reveal that Diarylheptanoids Vary in Amomum tsao-ko Fruit Development

Amomum tsao-ko is an important spice and medicinal plant that has received extensive attention in recent years for its high content of bioactive constituents with the potential for food additives and drug development. Diarylheptanoids are major and characteristic compounds in A. tsao-ko; however, the biochemical and molecular foundation of diarylheptanoids in fruit is unknown. We performed comparative metabolomics and transcriptomics studies in the ripening stages of A. tsao-ko fruit. The chemical constituents of fruit vary in different harvest periods, and the diarylheptanoids have a trend to decrease or increase with fruit development. GO enrichment analysis revealed that plant hormone signaling pathways including the ethylene-activated signaling pathway, salicylic acid, jasmonic acid, abscisic acid, and response to hydrogen peroxide were associated with fruit ripening. The biosynthetic pathways including phenylpropanoid, flavonoids, and diarylheptanoids biosynthesis were displayed in high enrichment levels in ripening fruit. The molecular networking and phytochemistry investigation of A. tsao-ko fruit has isolated and identified 10 diarylheptanoids including three new compounds. The candidate genes related to diarylheptanoids were obtained by coexpression network analysis and phylogenetic analysis. Two key genes have been verified to biosynthesize linear diarylheptanoids. This integrative approach provides gene regulation and networking associated with the biosynthesis of characteristic diarylheptanoids, which can be used to improve the quality of A. tsao-ko as food and medicine.

Flavonoids in Amomum tsaoko Crevost et Lemarie Ameliorate Loperamide-Induced Constipation in Mice by Regulating Gut Microbiota and Related Metabolites.

Amomum tsaoko (AT) is a dietary botanical with laxative properties; however, the active ingredients and mechanisms are still unclear. The active fraction of AT aqueous extract (ATAE) for promoting defecation in slow transit constipation mice is the ethanol-soluble part (ATES). The total flavonoids of ATES (ATTF) were the main active component. ATTF significantly increased the abundance of Lactobacillus and Bacillus and reduced the dominant commensals, such as Lachnospiraceae, thereby changing the gut microbiota structure and composition. Meanwhile, ATTF changed the gut metabolites mainly enriched in pathways such as the serotonergic synapse. In addition, ATTF increased the serum serotonin (5-HT) content and mRNA expression of 5-hydroxytryptamine receptor 2A (5-HT2A), Phospholipase A2 (PLA2), and Cyclooxygenase-2 (COX2), which are involved in the serotonergic synaptic pathway. ATTF increased Transient receptor potential A1 (TRPA1), which promotes the release of 5-HT, and Myosin light chain 3(MLC3), which promotes smooth muscle motility. Notably, we established a network between gut microbiota, gut metabolites, and host parameters. The dominant gut microbiota Lactobacillus and Bacillus, prostaglandin J2 (PGJ2) and laxative phenotypes showed the most significant associations. The above results suggest that ATTF may relieve constipation by regulating the gut microbiota and serotonergic synaptic pathway and has great potential for laxative drug development in the future.