CmMYC2-CmMYBML1 module orchestrates the resistance to herbivory by synchronously regulating the trichome development and constitutive terpene biosynthesis in Chrysanthemum.

Trichomes are specialized epidermal outgrowths covering the aerial parts of most terrestrial plants. There is a large species variability in occurrence of different types of trichomes such that the molecular regulatory mechanism underlying the formation and the biological function of trichomes in most plant species remain unexplored. Here, we used Chrysanthemum morifolium as a model plant to explore the regulatory network in trichome formation and terpenoid synthesis and unravel the physical and chemical roles of trichomes in constitutive defense against herbivore feeding. By analyzing the trichome-related genes from transcriptome database of the trichomes-removed leaves and intact leaves, we identified CmMYC2 to positively regulate both development of T-shaped and glandular trichomes as well as the content of terpenoids stored in glandular trichomes. Furthermore, we found that the role of CmMYC2 in trichome formation and terpene synthesis was mediated by interaction with CmMYBML1. Our results reveal a sophisticated molecular mechanism wherein the CmMYC2-CmMYBML1 feedback inhibition loop regulates the formation of trichomes (non-glandular and glandular) and terpene biosynthesis, collectively contributing to the enhanced resistance to Spodoptera litura larvae feeding. Our findings provide new insights into the novel regulatory network by which the plant synchronously regulates trichome density for the physical and chemical defense against herbivory.

Identification and functional analysis of terpene synthases revealing the secrets of aroma formation in Chrysanthemum aromaticum.

C. aromaticum is widely cultivated for its aromatic, medicinal, and tea-applicable properties, earning the nickname 'lavender in composite'. Terpenoids are the major compounds of C. aromaticum fragrance. To reveal the molecular mechanisms of terpenoid biosynthesis in C. aromaticum, NGS and SMRT sequencing were employed to identify the key terpene synthase genes. A total of 59,903 non-redundant transcripts were obtained by the transcriptome analysis. Twenty-nine terpene synthase genes (TPSs) were identified, and phylogenetic analysis showed that they belong to four subfamilies of terpene synthases. Five CaTPSs were successfully cloned. Subcellular localization showed they were present in the nucleus and cytosol. Structure models of five terpene synthases were predicted, and molecular docking results showed good binding affinities with FPP/GPP. In vitro enzymatic tests showed that CaTPS7, CaTPS8, CaTPS10 and CaTPS20 could catalyze substrates to produce terpenoids. CaTPS7 and CaTPS20 were both able to effectively convert the precursor FPP into caryophyllene. CaTPS8 could convert FPP to trans-nerolidol and nerolidyl acetate, while CaTPS10 could convert FPP to elemene and aristolochene. This study lays the groundwork for further research to depict the metabolism network of terpenoid in C. aromaticum. These identical terpene synthase genes could be introduced into the cultivated chrysanthemums to enhance their fragrance.

Integrated metabolomic and transcriptomic analysis reveals biosynthesis mechanism of flavone and caffeoylquinic acid in chrysanthemum.

BACKGROUND: Chrysanthemum morifolium 'HangBaiJu', a popular medicinal and edible plant, exerts its biological activities primarily through the presence of flavones and caffeoylquinic acids (CQAs). However, the regulatory mechanism of flavone and CQA biosynthesis in the chrysanthemum capitulum remains unclear. RESULTS: In this study, the content of flavones and CQAs during the development of chrysanthemum capitulum was determined by HPLC, revealing an accumulation pattern with higher levels at S1 and S2 and a gradual decrease at S3 to S5. Transcriptomic analysis revealed that CmPAL1/2, CmCHS1/2, CmFNS, CmHQT, and CmHCT were key structural genes in flavones and CQAs biosynthesis. Furthermore, weighted gene co-expression correlation network analysis (WGCNA), k-means clustering, correlation analysis and protein interaction prediction were carried out in this study to identify transcription factors (TFs) associated with flavone and CQA biosynthesis, including MYB, bHLH, AP2/ERF, and MADS-box families. The TFs CmERF/PTI6 and CmCMD77 were proposed to act as upstream regulators of CmMYB3 and CmbHLH143, while CmMYB3 and CmbHLH143 might form a complex to directly regulate the structural genes CmPAL1/2, CmCHS1/2, CmFNS, CmHQT, and CmHCT, thereby controlling flavone and CQA biosynthesis. CONCLUSIONS: Overall, these findings provide initial insights into the TF regulatory network underlying flavones and CQAs accumulation in the chrysanthemum capitulum, which laid a theoretical foundation for the quality improvement of C. morifolium 'HangBaiJu' and the high-quality development of the industry.

Characterization of bZIP Transcription Factors in Transcriptome of Chrysanthemum mongolicum and Roles of CmbZIP9 in Drought Stress Resistance.

bZIP transcription factors play important roles in regulating plant development and stress responses. Although bZIPs have been identified in many plant species, there is little information on the bZIPs in Chrysanthemum. In this study, bZIP TFs were identified from the leaf transcriptome of C. mongolicum, a plant naturally tolerant to drought. A total of 28 full-length bZIP family members were identified from the leaf transcriptome of C. mongolicum and were divided into five subfamilies based on their phylogenetic relationships with the bZIPs from Arabidopsis. Ten conserved motifs were detected among the bZIP proteins of C. mongolicum. Subcellular localization assays revealed that most of the CmbZIPs were predicted to be localized in the nucleus. A novel bZIP gene, designated as CmbZIP9, was cloned based on a sequence of the data of the C. mongolicum transcriptome and was overexpressed in tobacco. The results indicated that the overexpression of CmbZIP9 reduced the malondialdehyde (MDA) content and increased the peroxidase (POD) and superoxide dismutase (SOD) activities as well as the expression levels of stress-related genes under drought stress, thus enhancing the drought tolerance of transgenic tobacco lines. These results provide a theoretical basis for further exploring the functions of the bZIP family genes and lay a foundation for stress resistance improvement in chrysanthemums in the future.

Heterologous Expression of Chrysanthemum TCP Transcription Factor CmTCP13 Enhances Salinity Tolerance in Arabidopsis.

Plant-specific TEOSINTE BRANCHED1/CYCLOIDEA/PROLIFERATING CELL FACTOR (TCP) proteins play critical roles in plant development and stress responses; however, their functions in chrysanthemum (Chrysanthemum morifolium) have not been well-studied. In this study, we isolated and characterized the chrysanthemum TCP transcription factor family gene CmTCP13, a homolog of AtTCP13. This gene encoded a protein harboring a conserved basic helix-loop-helix motif, and its expression was induced by salinity stress in chrysanthemum plants. Subcellular localization experiments showed that CmTCP13 localized in the nucleus. Sequence analysis revealed the presence of multiple stress- and hormone-responsive cis-elements in the promoter region of CmTCP13. The heterologous expression of CmTCP13 in Arabidopsis plants enhanced their tolerance to salinity stress. Under salinity stress, CmTCP13 transgenic plants exhibited enhanced germination, root length, seedling growth, and chlorophyll content and reduced relative electrical conductivity compared with those exhibited by wild-type (WT) plants. Moreover, the expression levels of stress-related genes, including AtSOS3, AtP5CS2, AtRD22, AtRD29A, and AtDREB2A, were upregulated in CmTCP13 transgenic plants than in WT plants under salt stress. Taken together, our results demonstrate that CmTCP13 is a critical regulator of salt stress tolerance in plants.

Time-Course Analysis and Transcriptomic Identification of a Group III ERF CmTINY2 Involved in Waterlogging Tolerance in Chrysanthemums × morifolium Ramat.

'Hangju' is a variety of Chrysanthemum × morifolium Ramat. with both edible and medicinal value, cultivated as a traditional Chinese medicine for four centuries. The cultivation of 'Hangju' is currently at risk due to waterlogging, yet there is a lack of comprehensive understanding regarding its response to waterlogging stress. This study compared the waterlogging-tolerant 'Hangju' variety Enhanced Waterlogging Tolerance (EWT) with the waterlogging-sensitive variety CK ('zaoxiaoyangju'). EWT exhibited a more developed aeration tissue structure and demonstrated rapid growth regarding the adventitious roots following waterlogging. The time-course transcriptome analysis indicated that EWT could swiftly adjust the expression of the genes involved in the energy metabolism signaling pathways to acclimate to the waterlogged environment. Through WGCNA analysis, we identified Integrase-Type DNA-Binding Protein (CmTINY2) as a key factor in regulating the waterlogging tolerance in EWT. CmTINY2, a transcription factor belonging to the ethylene-responsive factor (ERF) subfamily III, operated within the nucleus and activated downstream gene expression. Its role in enhancing the waterlogging tolerance might be linked to the control of the stomatal aperture via the Ethylene-Responsive Element (ERE) gene. In summary, our research elucidated that the waterlogging tolerance displayed by EWT is a result of a combination of the morphological structure and molecular regulatory mechanisms. Furthermore, the study of the functions of CmTINY2 from ERF subfamily III also broadened our knowledge of the role of the ERF genes in the waterlogging signaling pathways.

Review of polysaccharides from Chrysanthemum morifolium Ramat.: Extraction, purification, structural characteristics, health benefits, structural-activity relationships and applications.

Chrysanthemum morifolium Ramat. (C. morifolium), as a traditional ornamental plant, it has multiple values, including edible, economic, nutritional and even medicinal values, which is used as herbal medicine and a new food resource in the world. Polysaccharides are one of the main bioactive components in C. morifolium, which have various health benefits such as improving functional constipation, improving colitis, anti-glycosylation, antioxidant, anti-angiogenesis, immunomodulation, prebiotic, and α-glucosidase inhibitory activities. This paper describes the extraction, purification, structural characteristics, health benefits, structural-activity relationships, applications, and analyses the shortcomings of the major relevant studies exist on C. morifolium polysaccharides. In addition, the potential mechanisms of the health benefits of C. morifolium polysaccharides were summarized. This study can provide reference and direction for further research and development of C. morifolium polysaccharides.

Buddleoside-rich Chrysanthemum indicum L. extract modulates macrophage-mediated inflammation to prevent metabolic syndrome induced by unhealthy diet.

BACKGROUND: Metabolic syndrome (MetS) is a precursor to the development of many diseases (atherosclerosis, diabetes, etc.). It is marked by disruptions in glucose and lipid metabolism, along with hypertension. Numerous types of risk factors contribute to the development of the MetS, inflammation and insulin resistance are present throughout the metabolic abnormalities. Chrysanthemum indicum L. is a traditional Chinese plant used for both tea and medicine, known for its high content of total flavonoids, which are important secondary metabolites. Our research led to the extraction of a Buddleoside-Rich Chrysanthemum indicum L. extract (BUDE) which has demonstrated anti-inflammatory properties. Nonetheless, the specific role and mechanism of BUDE in preventing MetS remain unclear. METHODS: The study initially evaluated the role of BUDE in preventing MetS. Subsequently, it investigated the anti-inflammatory properties of BUDE in the liver and pancreas in response to unhealthy diets. It then examined the level of insulin resistance and pancreatic ß-cell function induced by inflammation. Additionally, an lipopolysaccharide (LPS)-induced macrophage inflammation model was used to further investigate the ameliorative effects of BUDE in inflammation. RESULTS: BUDE has hypotensive, hypoglycemic and hypolipidemic effects. It can also resolve the imbalance between macrophage subpopulations, impede the triggering of the NF-κB signaling pathway, reduce the secretion of inflammatory mediators, ameliorate insulin resistance, and safeguard organs such as the liver and pancreas from inflammatory damage. These effects collectively contribute to preventing the development of MetS. DISCUSSION: BUDE has the ability to modulate macrophage-mediated inflammation, leading to improved insulin resistance. Additionally, it delivers antihypertensive, hypoglycemic, and hypolipidemic effects, offering a potential for preventing MetS.

Study of the Pharmacodynamic Material Basis And Mechanisms of the Action of Fubai Chrysanthemum in Relieving Visual Fatigue.

Fubai chrysanthemum is a kind of traditional Chinese medicine, which can be used as a common food, and is commonly used to improve and relieve visual fatigue. However, its pharmacodynamic material basis and action mechanisms in relieving visual fatigue have not been systematically studied. In this article, 11 absorbed ingredients from Fubai chrysanthemum were detected in rat plasma. Then, the target network pharmacology and KEGG pathway analysis were performed. It was found that Fubai chrysanthemum could inhibit various apoptotic cells and reduce oxidative damage of eyes by regulating the apoptosis pathway, thus alleviating visual fatigue. Further in vitro experiments showed that Fubai chrysanthemum could effectively protect against oxidation damage of adult retinal pigment epithelial cells (ARPE-19), retinal ganglion cells (RGC-5), and lens. The results of cell experiments showed that Fubai chrysanthemum could increase the cell activity, GSH content, and SOD content of ARPE-19 and RGC-5 after oxidative injury, while decreasing the IL-18 content. Similarly, in the study of lens transparency, we found that Fubai chrysanthemum could effectively alleviate the oxidative damage degree of the lens, and significantly increase the content of CAT, GSH, and SOD. The above results suggested that Fubai chrysanthemum could play an important role in alleviating visual fatigue through regulating cell apoptosis and antioxidative damage.

Identification of SDG gene family members and exploration of flowering related genes in different cultivars of chrysanthemums and their wild ancestors.

The SET domain genes (SDGs) are significant contributors to various aspects of plant growth and development, mainly includes flowering, pollen development, root growth, regulation of the biological clock and branching patterns. To clarify the biological functions of the chrysanthemum SDG family, the SDG family members of four chrysanthemum cultivars and three related wild species were identified; their physical and chemical properties, protein domains and conserved motifs were predicted and analyzed. The results showed that 59, 67, 67, 102, 106, 114, and 123 SDGs were identified from Chrysanthemum nankingense, Chrysanthemum lavandulifolium, Chrysanthemum seticuspe, Chrysanthemum × morifolium cv. 'Hechengxinghuo', 'Zhongshanzigui', 'Quanxiangshuichang' and 'Jinbeidahong', respectively. The SDGs were divided into 5-7 subfamilies by cluster analysis; different conserved motifs were observed in particular families. The SDGs of C. lavandulifolium and C. seticuspe were distributed unevenly on 9 chromosomes. SDG promoters of different species include growth and development, photo-response, stress response and hormone responsive elements, among them, the cis-acting elements related to MeJA response had the largest proportion. The expression of chrysanthemum SDG genes was observed for most variable selected genes which has close association with important Arabidopsis thaliana genes related to flowering regulation. The qPCR results showed that the expression trend of SDG genes varied in different tissues at different growth stages with high expression in the flowering period. The ClSDG29 showed higher expression in the flower and bud tissues, which indicate that ClSDG29 might be associated with flowering regulation in chrysanthemum. In summary, the results of this study can provide a basis for subsequent research on chrysanthemum flowering time regulation.

The Influence of Planting Sites On the Chemical Compositions of Chrysanthemum morifolium Flowers (Chuju) as Revealed by Py-GC/MS Combined with Multivariate Statistical Analysis.

Chuju, a cultivar of Chrysanthemum morifolium, has been traditionally cultivated for over 2000 years in China for both ornamental and medicinal purposes. To date, investigations into the chemical composition of this plant have indicated that it contains compounds with extensive biological activities, although detailed information on the chemical composition of Chuju remains scarce. In the present study, the chemical compositions of Chuju flowers were investigated across five sites in the core Chuju planting area in Anhui province, China. Analytical pyrolysis-gas chromatography/mass spectrometry (Py-GC/MS) was used to explore variations in flower chemical fingerprints from different Chuju planting sites. The study identified approximately 200 components in Chuju flowers and stems, including high levels of fatty acids, lipids, polysaccharides and terpenoids. Multivariate statistical analysis indicated that 16 chemical compounds were influential determinants of the chemical fingerprint and could be used to distinguish two clusters in the five core planting areas. The established Py-GC/MS analytical workflow could provide a basis for determining the chemical fingerprints of Chuju and help elucidate that products contain a reproducible content of bioactive compounds and overall quality for potential development of health and medicinal purposes.

Chrysanthemum lavandulifolium homolog ClCYCA2;1 modulates cell division in ray florets.

Morphology of ray florets in chrysanthemums is tightly associated with cell division and cell expansion, both of which require proper cell cycle progression. Here we identified a Chrysanthemum lavandulifolium homolog ClCYCA2;1, whose expression in ray florets is negatively correlated with petal width in C. lavandulifolium. Two TCP transcription factors in CYCLOIDEA2 (CYC2) family, ClCYC2a interacts with and stabilizes ClCYC2b and the latter can bind to the promoter of ClCYCA2;1 to activate its transcription. Overexpression of ClCYCA2;1 in C. lavandulifolium reduces the size of capitula and ray florets. Cytological analysis reveals that ClCYCA2;1 overexpression inhibits both cell division and cell expansion via repressing mitotic cell cycle in ray florets whose latitudinal development was more negatively influenced leading to increased ratios of petal length to width at later developmental stages. Yeast two hybrid library screening reveals multiple ClCYCA2;1 interacting proteins including ARP7, and silencing ClARP7 inhibits the development of ray florets. Co-immunoprecipitation assays confirm that ClCYCA2;1 can induce the degradation of ClARP7 to inhibit the development of ray florets. Taken together, our study constitutes a regulatory network containing ClCYC2b-ClCYCA2;1-ClARP7 in ray floret development via governing mitosis, which may facilitate breeding efforts targeted for novel ornamental traits of chrysanthemums.

Floral Response to Heat: A Study of Color and Biochemical Adaptations in Purple Chrysanthemums.

Chrysanthemums are among the world's most popular cut flowers, with their color being a key ornamental feature. The formation of these colors can be influenced by high temperatures. However, the regulatory mechanisms that control the fading of chrysanthemum flower color under high-temperature stress remain unclear. This study investigates the impact of high temperatures on the color and biochemical responses of purple chrysanthemums. Four purple chrysanthemum varieties were exposed to both normal and elevated temperature conditions. High-temperature stress elicited distinct responses among the purple chrysanthemum varieties. 'Zi Feng Che' and 'Chrystal Regal' maintained color stability, whereas 'Zi Hong Tuo Gui' and 'Zi lian' exhibited significant color fading, particularly during early bloom stages. This fading was associated with decreased enzymatic activities, specifically of chalcone isomerase (CHI), dihydroflavonol 4-reductase (DFR), and anthocyanidin synthase (ANS), indicating a critical period of color development under heat stress. Additionally, the color fading of 'Zi Lian' was closely related to the increased activity of the peroxidase (POD) and polyphenol oxidase (PPO). Conversely, a reduction in ß-glucosidase (ßG) activity may contribute significantly to the color steadfastness of 'Zi Feng Che'. The genes Cse_sc027584.1_g010.1 (PPO) and Cse_sc031727.1_g010.1 (POD) might contribute to the degradation of anthocyanins in the petals of 'Zi Hong Tuo Gui' and 'Zi Lian' under high-temperature conditions, while simultaneously maintaining the stability of anthocyanins in 'Zi Feng Che' and 'Chrystal Regal' at the early bloom floral stage. The findings of this research provide new insights into the physiological and biochemical mechanisms by which chrysanthemum flower color responds to high-temperature stress.

Identification of chrysanthemum variety via hyperspectral imaging and wavelength selection based on multitask particle swarm optimization.

Chrysanthemum, a widely favored flower tea, contains numerous phytochemicals for health benefits. Due to the different geographical origins and processing technics, its variety has a direct influence on the phytochemical content and pharmacological effect. Accordingly, an accurate identification for chrysanthemum varieties is significant for quality detection and market supervision. In this study, the hyperspectral imaging (HSI) combined with chemometrics methods was exploited to identify the chrysanthemum varieties. First, to alleviate the problem of easily trapping into local optimum in traditional spectral variable selection methods, the multi-tasking particle swarm optimization (MTPSO) was developed to select the key wavelengths by dividing hundreds of variables into low-dimensional subtasks. Second, to enrich the feature information, the spatial texture and color features contained in hyperspectral images were extracted and applied to chrysanthemum identification for the first time. Finally, an ensemble learning model, extreme gradient boosting (XGBoost), was constructed to conduct the chrysanthemum variety classification due to its strong generalization ability. Experimental results showed that the proposed MTPSO achieved the identification accuracy of 96.89%, and increased by 1.11-5.91% than classical spectral feature selection methods. Furthermore, after the involvement of spatial image information, the classification accuracy using spatial-spectral features was improved further, and reached 98.39%. Overall, this study highlights that the feature fusion of key wavelengths and spatial information is more effective for chrysanthemum variety identification, and can also provide technical reference for other HSI-related applications.

Integrated Metabolomics and Transcriptomics Reveal the Key Role of Flavonoids in the Cold Tolerance of Chrysanthemum.

Chrysanthemum (Chrysanthemum morifolium, ground-cover Chrysanthemums), one of the important garden flowers, has a high ornamental and economic value. However, its ornamental value is significantly diminished by the low temperature experienced in northeastern China. Here, metabolomics and transcriptomics were performed on three Chrysanthemum cultivars before and after a low temperature to investigate the dynamic metabolite changes and the molecular regulatory mechanisms. The results showed that 1324 annotated metabolites were detected, among which 327 were identified as flavonoids derived from Chrysanthemum. The accumulation of metabolites and gene expression related to the flavonoid biosynthesis pathway significantly increased in the three cultivars under the low temperature, indicating flavonoid metabolism actively participates in the Chrysanthemum cold response. Specifically, the content of cyanidin and pelargonidin derivatives and the expression of anthocyanin biosynthesis genes significantly increases in XHBF, providing a reasonable explanation for the change in petal color from white to purple under the low temperature. Six candidate UDP-glycosyltransferase genes involved in the glycosylation of flavonoids were identified through correlation networks and phylogenetic analysis. CmNAC1, CmbZIP3, and other transcription factors potentially regulating flavonoid metabolism and responding to low temperatures were discovered by correlation analysis and weighted gene co-expression network analysis (WGCNA). In conclusion, this study elucidated the specific response of flavonoids to low temperatures in Chrysanthemums, providing valuable insights and metabolic data for investigating cold tolerance.

Prevention of supercritical carbon dioxide fluid extract from Chrysanthemum indicum Linnén on cutaneous squamous cell carcinomas progression following UV irradiation in mice.

Chrysanthemum indicum Linnén (C. indicum), a medicinal and food herb with various bioactive components, may be of beneficial use in cosmetics and the treatment of skin-related diseases. However, to date, few studies have been reported on its potential preventive and therapeutic effects on skin cancer. Therefore, the present study aimed to investigate the effect and potential mechanism of action of supercritical carbon dioxide extract from C. indicum (CISCFE) on UV-induced skin cancer in a mouse model. Kunming mice were allocated randomly to five treatment groups: Sham, model, low concentration CISCFE, high concentration CISCFE and positive control nicotinamide groups. The dorsal skin of mice was irradiated with UV light for 31 weeks. Histopathological changes, ELISA assays, immunohistochemical analysis and western blotting were performed to investigate the potential therapeutic effects of CISCFE. The results showed that CISCFE alleviated skin oxidative and inflammatory damage in a UV-induced mouse model of skin cancer. Moreover, CISCFE suppressed abnormal activation of proto-oncogene c-Myc and the overexpression of Ki-67 and VEGF, and increased expression of the anti-oncogene PTEN, thereby reducing abnormal proliferation of the epidermis and blood vessels. Additionally, CISCFE increased the protein expression levels of NAD-dependent protein deacetylase sirtuin-1 (SIRT1), Kelch-like ECH associated protein 1 (Keap1) and inhibited the expression of nuclear factor 2 erythroid 2-related factor 2 (Nrf2), phosphorylated (p)-p62 (Ser 349), p-p65 and acetyl-p65 proteins in a UV-induced skin cancer mouse model. In summary, CISCFE exhibited potent anti-skin cancer activity, which may be attributed its potential effects on the p62/Keap1-Nrf2 and SIRT1/NF-κB pathways.

Research on extracting and preparing a Puerariae Flos and Chrysanthemum-based drink.

In traditional Chinese medicine, Puerariae Flos and Chrysanthemum are widely utilized in herbal teas for hangover relief and heat-clearing detoxification. In this study, a new drink has been developed, employing these two flowers as primary raw materials. The objective of this study was to optimize the optimal formula, extraction process, and preparation method for the drink. The optimization of the formula and extraction process was guided by the utilization of the total flavonoids content in the water decoction of the two flowers as an indicator. Based on the sensory evaluation criteria, including color, smell, taste, and state of the drink, the water decoction addition, honey addition, and citric acid addition were optimized by single-factor experiments and orthogonal experiments. The best formula and extraction process was 10 g of Puerariae Flos, 10 g of Chrysanthemum, 48 min of decocting time, and 615 mL of water. The optimal preparation process consisted of 30% water decoction, 8% honey, and 0.025% citric acid. Subsequently, a golden yellow, transparent, and stable liquid was produced, possessing a sweet taste along with the distinctive aroma and flavor of Puerariae Flos and Chrysanthemum. Supplementary Information: The online version contains supplementary material available at 10.1007/s13197-024-05937-x.

Chrysanthemum morifolium Ramat extract and probiotics combination ameliorates metabolic disorders through regulating gut microbiota and PPARα subcellular localization.

BACKGROUND: Chrysanthemum morifolium Ramat, a traditional Chinese medicine, has the effects on liver clearing, vision improving, and anti-inflammation. C. morifolium and probiotics have been individually studied for their beneficial effects on metabolic diseases. However, the underlying molecular mechanisms were not completely elucidated. This study aims to elucidate the potential molecular mechanisms of C. morifolium and probiotics combination (CP) on alleviating nonalcoholic fatty liver disease (NAFLD) and the dysregulation of glucose metabolism in high-fat diet (HFD)-fed mice. METHODS: The therapeutic effect of CP on metabolism was evaluated by liver histology and serum biochemical analysis, as well as glucose tolerance test. The impact of CP on gut microbiota was analyzed by 16S rRNA sequencing and fecal microbiota transplantation. Hepatic transcriptomic analysis was performed with the key genes and proteins validated by RT-qPCR and western blotting. In addition, whole body Pparα knockout (Pparα-/-) mice were used to confirm the CP-mediated pathway. RESULTS: CP supplementation ameliorated metabolic disorders by reducing body weight and hepatic steatosis, and improving glucose intolerance and insulin resistance in HFD fed mice. CP intervention mitigated the HFD-induced gut microbiota dysbiosis, which contributed at least in part, to the beneficial effect of improving glucose metabolism. In addition, hepatic transcriptomic analysis showed that CP modulated the expression of genes associated with lipid metabolism. CP downregulated the mRNA level of lipid droplet-binding proteins, such as Cidea and Cidec in the liver, leading to more substrates for fatty acid oxidation (FAO). Meanwhile, the expression of CPT1α, the rate-limiting enzyme of FAO, was significantly increased upon CP treatment. Mechanistically, though CP didn't affect the total PPARα level, it promoted the nuclear localization of PPARα, which contributed to the reduced expression of Cidea and Cidec, and increased expression of CPT1α, leading to activated FAO. Moreover, whole body PPARα deficiency abolished the anti-NAFLD effect of CP, suggesting the importance of PPARα in CP-mediated beneficial effect. CONCLUSION: This study revealed the hypoglycemic and hepatoprotective effect of CP by regulating gut microbiota composition and PPARα subcellular localization, highlighting its potential for therapeutic candidate for metabolic disorders.

Influence of planting dates and fertilizer modules on yield of chrysanthemum and soil health.

BACKGROUND: Optimum planting date and appropriate fertilizer module are essential facets of chrysanthemum cultivation, to enhance quality yield, and improve soil health. A field-based study was undertaken over multiple growing seasons in 2022 and 2023, where six different planting dates, viz., P1:June 15, P2:June 30, P3:July 15, P4:July 30, P5:August 15 and P6:August 30 and two fertilizer modules, FM1:Jeevamrit @ 30 ml plant-1 and FM2:NPK @ 30 g m-2 were systematically examined using a Randomized Block Design (factorial), replicated thrice. RESULTS: P6 planting resulted in early bud formation (44.03 days) and harvesting stage (90.78 days). Maximum plant height (79.44 cm), plant spread (34.04 cm), cut stem length (68.40 cm), flower diameter (7.83 cm), stem strength (19.38˚), vase life (14.90 days), flowering duration (24.08 days), available soil N (314 kg ha-1), available P (37 kg ha-1), available K (347 kg ha-1), bacterial count (124.87 × 107 cfu g-1 soil), actinomycetes count (60.72 × 102 cfu g-1 soil), fungal count (30.95 × 102 cfu g-1 soil), microbial biomass (48.79 µg g-1 soil), dehydrogenase enzyme (3.64 mg TPF h-1 g-1 soil) and phosphatase enzyme (23.79 mol PNP h-1 g-1 soil) was recorded in P1 planting. Among the fertilization module, minimum days to bud formation (74.94 days) and days to reach the harvesting stage (120.95 days) were recorded with the application of NPK @30 g m-2. However, maximum plant height (60.62 cm), plant spread (23.10 cm), number of cut stems m-2 (43.88), cut stem length (51.34 cm), flower diameter (6.92 cm), stem strength (21.24˚), flowering duration (21.75 days), available soil N (317 kg ha-1), available P (37 kg ha-1) and available K (349 kg ha-1) were also recorded with the application of NPK @300 kg ha-1. Maximum vase life (13.87 days), OC (1.13%), bacterial count (131.65 × 107 cfu g-1 soil), actinomycetes count (60.89 × 102 cfu g-1 soil), fungal count (31.11 × 102 cfu g-1 soil), microbial biomass (51.27 µg g-1 soil), dehydrogenase enzyme (3.77 mg TPF h-1 g-1 soil) and phosphatase enzyme (21.72 mol PNP h-1 g-1 soil) were observed with the application of Jeevamrit @ 30 ml plant-1. CONCLUSION: Early planting (P1) and inorganic fertilization (NPK @ 30 g m-2) resulted in improved yield and soil macronutrient content. The soil microbial population and enzymatic activity were improved with the jeevamrit application. This approach highlights the potential for improved yield and soil health in chrysanthemum cultivation, promoting a more eco-friendly and economically viable agricultural model.

An improved genome assembly of Chrysanthemum nankingense reveals expansion and functional diversification of terpene synthase gene family.

BACKGROUND: Terpenes are important components of plant aromas, and terpene synthases (TPSs) are the key enzymes driving terpene diversification. In this study, we characterized the volatile terpenes in five different Chrysanthemum nankingense tissues. In addition, genome-wide identification and expression analysis of TPS genes was conducted utilizing an improved chromosome-scale genome assembly and tissue-specific transcriptomes. The biochemical functions of three representative TPSs were also investigated. RESULTS: We identified tissue-specific volatile organic compound (VOC) and volatile terpene profiles. The improved Chrysanthemum nankingense genome assembly was high-quality, including a larger assembled size (3.26 Gb) and a better contig N50 length (3.18 Mb) compared to the old version. A total of 140 CnTPS genes were identified, with the majority representing the TPS-a and TPS-b subfamilies. The chromosomal distribution of these TPS genes was uneven, and 26 genes were included in biosynthetic gene clusters. Closely-related Chrysanthemum taxa were also found to contain diverse TPS genes, and the expression profiles of most CnTPSs were tissue-specific. The three investigated CnTPS enzymes exhibited versatile activities, suggesting multifunctionality. CONCLUSIONS: We systematically characterized the structure and diversity of TPS genes across the Chrysanthemum nankingense genome, as well as the potential biochemical functions of representative genes. Our results provide a basis for future studies of terpene biosynthesis in chrysanthemums, as well as for the breeding of improved chrysanthemum varieties.