Lactobacillus casei-fermented Amomum xanthioides ameliorates metabolic dysfunction in high-fat diet-induced obese mice.

Amomum xanthioides (AX) has been used as an edible herbal medicine to treat digestive system disorders in Asia. Additionally, Lactobacillus casei is a well-known probiotic commonly used in fermentation processes as a starter. The current study aimed to investigate the potential of Lactobacillus casei-fermented Amomum xanthioides (LAX) in alleviating metabolic disorders induced by high-fat diet (HFD) in a mouse model. LAX significantly reduced the body and fat weight, outperforming AX, yet without suppressing appetite. LAX also markedly ameliorated excessive lipid accumulation and reduced inflammatory cytokine (IL-6) levels in serum superior to AX in association with UCP1 activation and adiponectin elevation. Furthermore, LAX noticeably improved the levels of fasting blood glucose, serum insulin, and HOMA-IR through positive regulation of glucose transporters (GLUT2, GLUT4), and insulin receptor gene expression. In conclusion, the fermentation of AX demonstrates a pronounced mitigation of overnutrition-induced metabolic dysfunction, including hyperlipidemia, hyperglycemia, hyperinsulinemia, and obesity, compared to non-fermented AX. Consequently, we proposed that the fermentation of AX holds promise as a potential candidate for effectively ameliorating metabolic disorders.

[Multi-index evaluation of different parts of Amomum villosum].

To investigate the quality differences between the seeds and husks of Amomum villosum and explore the rationality of using the seeds without husks, this study determined the content of protocatechuic acid, vanillic acid, epicatechin, quercitrin, volatile oil, water extract, and ethanol extract. The 2,2-diphenyl-1-picrylhydrazyl(DPPH), 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)(ABTS), and hydroxyl radical scavenging activities were determined to evaluate the antioxidant activities of seeds and husks. The quality differences between the seeds and husks were assessed through orthogonal partial least squares-discriminant analysis(OPLS-DA) and analytic hierarchy process(AHP) combined with the entropy weight method(EWM). Significant differences(P<0.05) were observed in all 10 indicators between the seeds and husks. The levels of epicatechin, quercetin, and volatile oil were higher in the seeds, whereas those of protocatechuic acid, vanillic acid, water extract, and ethanol extract were higher in the husks. The seeds showed stronger scavenging ability against DPPH and ABTS radicals, while the husks showed a stronger scavenging effect on hydroxyl radicals. OPLS-DA significantly discriminated between the seeds and husks. Furthermore, volatile oil, water extract, DPPH radical scavenging rate, quercitrin, ABTS radical scavenging rate, hydroxyl radical scavenging rate, and vanillic acid were selected as the main differential indicators by variable importance in projection(VIP). Comprehensive scores calculated by AHP combined with EWM indicated that the seeds were superior to husks in terms of overall quality. However, there are still some dominant components and a certain antioxidant effect in the husks. Therefore, it is suggested to using Amomi Fructus with a certain amount of husks or utilizing the husks for other purposes.

Lactobacillus Casei-fermented Amomum Xanthioides Mitigates non-alcoholic fatty liver disease in a high-fat diet mice model.

Non-alcoholic fatty liver disease (NAFLD) is a substantial global health issue owing to its high prevalence and the lack of effective therapies. Fermentation of medicinal herbs has always been considered a feasible strategy for enhancing efficacy in treating various ailments. This study aimed to investigate the potential benefits of the Lactobacillus casei-fermented Amomum xanthioides (LAX) on NAFLD in a high-fat diet model. HFD-fed C57BL6/j mice were administered with 200 mg/kg of LAX or unfermented Amomum xanthioides (AX) or 100 mg/kg of metformin for 6 weeks from the 4th week. The 10-week HFD-induced alterations of hepatic lipid accumulation and hepatic inflammation were significantly attenuated by LAX dominantly (more than AX or metformin), which evidenced by pathohistological findings, lipid contents, inflammatory cytokines including tumor necrosis factor (TNF)-α, interleukin (IL)- 6 and IL-1ß, oxidative parameters such as reactive oxygen species (ROS) and malondialdehyde (MDA), and molecular changes reversely between lipogenic proteins such as glycerol-3-phosphate acyltransferase (GPAM) and sterol regulatory element-binding protein (SREBP)- 1, and lipolytic proteins including peroxisome proliferator-activated receptor (PPAR-α) and AMP-activated kinase (AMPK)-α in the liver tissues. In addition, the abnormal serum lipid parameters (triglyceride, total cholesterol and LDL-cholesterol) notably ameliorated by LAX. In conclusion, these findings support the potential of LAX as a promising plant-derived remedy for NAFLD.

Phenolics and terpenoids with good anti-inflammatory activity from the fruits of Amomum villosum and the anti-inflammatory mechanism of active diterpene.

The fruits of Amomum villosum are often considered a medicinal and food homologous material and have been found to have therapeutic effects in chronic enteritis, gastroenteritis, and duodenal ulcer. The aim of this study is to discover the anti-inflammatory active ingredients from dried ripe fruits of A. villosum and to elucidate the molecular mechanisms. We verified that the inhibitory activity of the ethyl acetate extract was superior to Dexamethasone (Dex), so we ultimately chose to study the ethyl acetate extract from the fruits of A. villosum. A total of 33 compounds were isolated from its ethyl acetate extract, including nine known diterpenoids (compounds 1-9), twelve known sesquiterpenoids (compounds 10-21), ten known phenolics (compounds 22, 23, 25-29, 31-33) and two new phenolics (24 and 30). On the basis of chemical evidences and spectral data analysis (UV, ECD, Optical rotation data, 1D and 2D-NMR, HR-ESI-MS, NMR chemical shift calculations), the structures of new compounds were elucidated. Among these compounds, isocoronarin D (5) was found to have good anti-inflammatory activity. Further research has found that isocoronarin D can down-regulate the protein levels of COX2 and NOS2, activate Nrf2/Keap1 and suppress NF-κB signaling pathway in LPS-induced RAW264.7 cells. In addition, isocoronarin D inhibited inflammasome assembly during inflammasome activation by hampering the binding of NLRP3 and ASC. Further evidence revealed that isocoronarin D suppressed the assembly of the NLRP3 inflammasome via blocking the formation of ASC specks. From these results, isocoronarin D may be the important bioactive compound of A. villosum and exhibits anti-inflammatory effects by regulating the NF-κB/Nrf2/NLRP3 axis in macrophages.

Integrating chemical similarity and bioequivalence: an overall evaluation of the quality consistency of traditional decoction and dispensing granule decoction of Amomum villosum.

OBJECTIVE: This study aims to investigate the quality consistency between traditional decoction (TD) of Amomum villosum and its dispensing granule decoction (DGD). Fifteen batches of TD and nine batches of dispensing granules (manufactured by A, B, and C) were prepared and evaluated for their consistency. METHODS: Firstly, The chemical similarity of TD and DGD was examined using GC and HPLC, coupled with hierarchical cluster analysis (HCA), criteria importance though intercrieria correlation(CRITIC) weighting method, and principal component analysis (PCA). Secondly, the gastrointestinal motility experiments in mice, along with the CRITIC weighting method, were employed to assess the bioequivalence of TD and DGD of Amomum villosum. Finally, the entropy weight technique-gray relative analysis(GRA) method was used to compare the quality of Amomum villosum decoctions. RESULTS: ①The CRITIC weighting method indicated significantly higher scores for TD than DGD (p < 0.01). HCA and PCA results demonstrated a clear distinction between TD and DGD. ②Gastrointestinal motility test results revealed no significant difference between TD and DGD in other indicators (p > 0.05).③Gray relative analysis results showed that the relative correlation of TD was more significant than that of DGD. CONCLUSION: The chemical composition of DGD and TD differed. The biological activity of DGD-A/B was consistent with that of TD, while the difference between DGD-C and TD was significant. A comprehensive evaluation showed that TD exhibited better quality than DGD. DGD manufacturers should optimize the preparation process to enhance product quality.

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.

Network pharmacology combined with an animal model to reveal the material basis and mechanism of Amomum villosum in alleviating constipation in mice.

Constipation is a prevalent gastrointestinal disorder, with its prevalence showing an annual upward trend. There are many factors involved in the occurrence of constipation, such as abnormal smooth muscle contraction and disorders of gastrointestinal hormone secretion. Amomum villosum (A. villosum) has been proven to be effective in improving digestive system diseases, but there is no report on improving constipation. Therefore, we used network pharmacology prediction combined with animal experiments to explore the key active components of A. villosum and their pharmacological mechanisms. The results of network pharmacological prediction showed that ß-sitosterol was the key laxative compound of A. villosum, which may play a laxative role by activating the adrenoceptor alpha 1 A-myosin light chain (ADRA1A-MLC) pathway. Further animal experiments showed that ß-sitosterol could significantly shorten the time to first black stool; increase faecal weight, faecal number, and faecal water content; and promote gastrointestinal motility. ß-sitosterol may promote intestinal motility by upregulating the expression of ADRA1A and myosin light chain 9 (Myl9) mRNA and protein in the colon, thereby activating the ADRA1A-MLC signalling pathway. In addition, it is possible to improve constipation symptoms by regulating serum neurotransmitters and gastrointestinal motility-related factors, such as the serum content of 5-hydroxytryptamine (5-HT) and acetylcholinesterase (AchE) and the mRNA expression of 5-hydroxytryptamine receptor 4 (5-HT4), stem cell factor (SCF), stem cell factor receptor (c-Kit) and smooth muscle myosin light chain kinase (smMLCK) in the colon. These results lay a foundation for the application of A. villosum and ß-sitosterol in constipation.

Assessing heavy metal contamination in Amomum villosum Lour. fruits from plantations in Southern China: Soil-fungi-plant interactions.

Amomum villosum Lour. fruit is a common healthy food widely cultivated in southern China. Heavy metal contamination of farmland soils has becomes a serious environmental concern in China. Heavy metals in soil can be introduced into the food chain and pose health risks to humans. However, microbial communities may play beneficial roles in plants grown in metal-polluted soils. This study aimed to assess the potential health risks of heavy metals in soils and A. villosum fruits from different production areas and to explore the soil-microbe-plant regulation pattern for heavy metals in A. villosum fruits. Soil and A. villosum fruit samples were collected from nine planting fields in four provinces of southern China. The results showed that soils from seven areas were polluted with heavy metals to different degrees. Cr and Mn were the most serious contaminating elements. However, the accumulation of heavy metals in A. villosum fruit was negligible with no expected human health risks. Partial least squares path analysis of structural equation modeling showed that the accumulation of heavy metals in A. villosum fruits was influenced by multiple factors. More importantly, the PLS-SEM revealed that the heavy metal content in A. villosum fruits was indirectly affected by soil heavy metals through the regulation of the microbial community. Furthermore, some fungal phyla (e.g., Ascomycota and Chytridiomycota) and genera (e.g., Mucor) were related to the heavy metal content in the soil and in A. villosum fruits. The results of this study verified that soil fungal community play an important role in the accumulation of heavy metals in A. villosum fruits. Using fungi provides a potential biological strategy for reducing the health risk posed by heavy metals in food.

Black cardamom (Amomum subulatum) extract improves growth potential, antioxidant status, immune parameters and response to crowding stress in Catla catla.

Natural herbs are excellent alternatives to synthetic compounds to enhance the growth performance and health status of fish. This study was conducted to evaluate the efficacy of black cardamom (Amomum subulatum) extract (BCE) as an herbal supplement in improving the growth, antioxidant status, haematology, and serum biochemistry of Catla catla. The acclimatized fish (N = 900; average initial weight = 14.44 ± 0.33 g) were allocated into five groups (60 fish/group in triplicate) in hapas (4 × 2 × 2.5 ft) and provided with feed containing 0 (control), 0.5, 1, 2 and 4 g/kg BCE for 90 days before being subjected to 8 days of crowding stress. After 90 days, significantly higher feed utilization and growth were observed in all BCE-fed treatments compared to the control; however, the maximum values of these parameters were noted in the 2 g/kg BCE-fed treatment. Moreover, the BCE-fed groups exhibited a significant increase in antioxidant indices (glutathione peroxidase, superoxide dismutase and catalase), with a significant reduction in malondialdehyde levels, indicating a higher antioxidant capacity compared to the control. Significant improvements in haematological parameters, such as an increase in haematocrit, haemoglobin and red blood cells and a decrease in white blood cells, were observed in BCE-fed treatments compared to the control. Furthermore, BCE-fed groups showed a significant decrease in serum glucose, cortisol and triglycerides, while total protein, globulin and albumin levels were significantly higher compared with the control. During the feeding trial of 90 days, no mortality was observed; however, the lowest cumulative mortality was noted in the 2 g/kg BCE group after crowding stress of 8 days. In conclusion, the use of BCE in feed can enhance the growth performance, antioxidant status, haematology, and serum biochemistry of C. catla and improve the resistance against crowding stress. The optimum levels of BCE for C. catla were estimated based on weight gain % (1.78 g/kg; R2 = 0.97), feed conversion ratio (1.65 g/kg; R2 = 0.98), MDA content (1.66 g/kg; R2 = 0.93) and serum lysozyme activity (1.72 g/kg; R2 = 1) using broken-line regression analysis.

Elevation determines the productivity of large cardamom (Amomum subulatum Roxb.) cultivars in Sikkim Himalaya.

Large cardamom (Amomum subulatum Roxb.) is an economically important cash crop that provides a livelihood option for the rural communities in Sikkim Himalaya. However, its production has declined drastically over the past few decades due to climate change and other factors affecting the livelihood of marginal cardamom-dependent farmers in the region. Climate change causes a shift in elevational distributions of mountain species, and it is pivotal to understand its effect on yield and yield-related traits for economically important plant species like large cardamom. For this, we randomly studied 41 large cardamom cultivation sites in Sikkim (India) with elevations ranging between 975 and 2069 m asl and evaluated the yield-related traits (number of capsules per spike, capsule length, capsule width, fresh capsule weight, dry capsule weight, number of seeds per locule, fresh seed weight, and dry seed weight) in five cultivars (Dzongu Golsey, Sawney, Seremna, Ramsey, and Varlangey). We observed a significant variability (P < 0.05) for morphometric traits in each of the five cultivars cultivated in different elevations. The cultivation of low-elevation cultivars like Seremna and Dzongu Golsey (suitable in elevation < 975 m) has shifted upward to mid (975-1515 m) and high-elevation (> 1515 m), while cultivation of high-elevation Ramsey cultivar (suitable in elevation > 1515 m) has shifted downward (< 1515 m). The Dzongu Golsey, Sawney, and Seremna cultivated in mid-elevation (975-1515 m) performed better in terms of yield-related traits than the same cultivars cultivated in high-elevation (> 1515 m) and showed moderate to high negative correlation between elevation and yield-related traits, indicating the negative effect of elevation on their yield. Likewise, Ramsey and Varlangey cultivated in high elevation (> 1515 m) performed better than the one cultivated in mid-elevation (975-1515 m) and depicted moderate to high positive correlation between elevation and yield-related traits, suggesting a positive influence of elevation on their yield. Although there is an elevational shift in the cultivation of large cardamom cultivars, the elevation influences the performance of the large cardamom cultivars, and it also suggests cultivating the cultivars in their suitable elevation range for better productivity.

Chemical Composition of Essential Oils and Supercritical Carbon Dioxide Extracts from Amomum kravanh, Citrus hystrix and Piper nigrum 'Kampot'.

The fruits of Amomum kravanh, Citrus hystrix and Piper nigrum 'Kampot' are traditionally used as spices in Cambodian cuisine. In this study, the chemical composition of essential oils (EOs) and supercritical CO2 extracts from all three species was determined using GC-MS, with two columns of different polarity (HP-5/DB-HeavyWAX). Differences between the chemical profile of the EOs and CO2 extracts were observed for all species. The greatest difference was detected in A. kravanh EO containing mainly eucalyptol (78.8/72.6%), while the CO2 extract was rich in fatty acids (13/55.92%) and long-chain alkanes (25.55/9.54%). Furthermore, the results for the CO2 extract of this species differed, where tricosane (14.74%) and oleic acid (29.26%) were the main compounds identified when utilizing the HP-5 or DB-HeavyWAX columns, respectively. Moreover, the EO and CO2 extract from P. nigrum 'Kampot' fruits and the CO2 extract from C. hystrix fruit peel, containing respective amounts 34.84/39.55% (for EO) and 54.21/55.86% (for CO2 extract) of ß-caryophyllene and 30.2/28.9% of ß-pinene, were isolated and analyzed for the first time. Generally, these findings suggest that supercritical CO2 could potentially be used for the extraction of all three spices. Nevertheless, further research determining the most efficient extraction parameters is required before its commercial application.

Amomum subulatum fruits protect against radiation-induced esophagitis by regulating antioxidant status and inflammatory responses.

Radiation esophagitis (RE) is an inimical event that requires proper management while carrying out radiotherapy for thoracic cancers. The present study investigates the protective effect of dry fruits of the culinary and folkloric spice Amomum subulatum against experimental thoracic radiation-induced esophagitis. C57BL/6 mice were subjected to 25 Gy whole thorax irradiation and administered with 250 mg/kg body weight of methanolic extract of A. subulatum dry fruits (MEAS) orally for four consecutive weeks. Changes in tissue antioxidant activities, oxidative stress parameters, expression of antioxidant, inflammation, and fibrosis-related genes were observed. Administration of MEAS boosted antioxidant status, thereby reducing radiation-induced oxidative stress in the esophagus. PCR (polymerase chain reaction) results showed decreased expression of apoptosis, inflammation, and fibrosis-associated genes as well as increased expression of vital cytoprotective and antioxidant genes in MEAS-treated mice, manifesting its protective effect against radiation-induced oxidative stress, inflammatory responses, and fibrosis in the esophagus. Further, histopathology, immunohistochemistry (Cyclooxygenase-2), and Masson's Trichrome staining ascertained the protective effect of MEAS in alleviating radiation-induced esophageal injury. The synergistic effect of bioactive phytochemicals in MEAS with potent antioxidant and anti-inflammatory efficacies might have contributed to its mitigating effect against RE. Taken together, our results ascertained the radioprotective potential of MEAS, suggesting its possible nutraceutical application as a radiation countermeasure.

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

BACKGROUND: The 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. RESULTS: Fourier 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. SIGNIFICANCE: This 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.

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.

Three New Labdane-Type Diterpenoids from the Fruits of Amomum villosum and Their Anti-Inflammatory Activities.

Three new labdane-type diterpenoids, calcaratarin E, villosumtriol, and 12-epi-villosumtriol (1-3) were isolated from the fruits of Amomum villosum, along with seven known diterpenoids (4-10). Through comprehensive analysis of chemical evidence and spectral data including UV, 1D and 2D NMR, HR-ESI-MS, IR, and X-ray crystallography, the structures of these novel compounds were successfully determined. Additionally, the inhibitory effects of compounds 2-10 on NO production in lipopolysaccharide (LPS)-induced RAW264.7 cells were evaluated. Notably, compound 6 exhibited the most significant inhibitory effect with an IC50 value of 1.74±0.69 µM.

Chemical Constituents from the Fruits of Amomum kravanh and Their Role in Activating Alcohol Dehydrogenase.

Alcoholism is a worldwide health problem, and diseases caused by alcoholism are killing people every year. Amomum kravanh is a traditional Chinese medicine used to relieve hangovers. However, whether its bioactive components improve alcohol metabolism is not clear. In this study, ten new (amomumols A-J, 1-10) and thirty-five known (11-45) compounds were isolated from the fruits of Amomum kravanh by an activity-guided separation. Ten novel compounds were identified as four sesquiterpenoids (1-4), three monoterpene derivatives (5-7), two neolignans (8, 9), and a novel norsesquiterpenoid (10) with a new C14 nor-bisabolane skeleton. Their structures were determined by the comprehensive analysis of high-resolution electrospray ionization mass spectrometry (HRESIMS), nuclear magnetic resonance (NMR), and electronic circular dichroism (ECD) calculation. The effects of all isolated compounds on the activity of alcohol dehydrogenase were evaluated in vitro, and it was found that eight compounds (11, 12, 15, 18, 26, and 36-38) exhibited significant activation effects on the alcohol dehydrogenase at 50 µM.

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.

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.

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.

Transcriptome and proteome analyses reveal the potential mechanism of seed dormancy release in Amomum tsaoko during warm stratification.

BACKGROUND: In Amomum tsaoko breeding, the low germination rate is the major limitation for their large-scale reproduction. We found that warm stratification was an effective treatment to break the seed dormancy of A. tsaoko prior to sowing and could be an important component of improving breeding programs. The mechanism of seed dormancy release during warm stratification remains unclear. Therefore, we studied the differences between transcripts and proteomes at 0, 30, 60, and 90 days of warm stratification, to identify some regulatory genes and functional proteins that may cause seed dormancy release in A. tsaoko and reveal their regulatory mechanism. RESULTS: RNA-seq was performed for the seed dormancy release process, and the number of differentially expressed genes (DEGs) was 3196 in three dormancy release periods. Using TMT-labelling quantitative proteome analysis, a total of 1414 proteins were defined as differentially expressed proteins (DEPs). Functional enrichment analyses revealed that the DEGs and DEPs were mainly involved in signal transduction pathways (MAPK signaling, hormone) and metabolism processes (cell wall, storage and energy reserves), suggesting that these differentially expressed genes and proteins are somehow involved in response to seed dormancy release process, including MAPK, PYR/PYL, PP2C, GID1, GH3, ARF, AUX/IAA, TPS, SPS, and SS. In addition, transcription factors ARF, bHLH, bZIP, MYB, SBP, and WRKY showed differential expression during the warm stratification stage, which may relate to dormancy release. Noteworthy, XTH, EXP, HSP and ASPG proteins may be involved in a complex network to regulate cell division and differentiation, chilling response and the seed germination status in A. tsaoko seed during warm stratification. CONCLUSION: Our transcriptomic and proteomic analysis highlighted specific genes and proteins that warrant further study in fully grasping the precise molecular mechanisms that control the seed dormancy and germination of A. tsaoko. A hypothetical model of the genetic regulatory network provides a theoretical basis for overcoming the physiological dormancy in A. tsaoko in the future.