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With the increase of chemotherapy frequency for breast cancer, the drug resistance rate of patients is rising, accompanied by cell invasion and metastasis, thus causing mortality. We aimed to explore the mechanism by which Platycodon grandiflorus affects breast cancer cells in terms of the doxorubicin (Dox) resistance and epithelial-mesenchymal transition (EMT) via the p38 mitogen-activated protein kinase (p38 MAPK) signaling pathway. MCF-7/R cell lines with resistance to Dox were established. After 24 h of culture with DMEM (blank group), they were divided into Platycodon grandiflorus, Platycodon grandiflorus + Ophiopogon japonicus, Platycodon grandiflorus + Curcumae Rhizoma, Platycodon grandiflorus + Curcumae Rhizoma + U46619 groups. Flow cytometry, colony formation assay, as well as Transwell assay were performed to examine the cells for apoptosis, proliferation, and invasion, respectively. Western blotting was performed to measure the phosphorylated (p)-p38 MAPK-to-p38 MAPK ratio together with N-cadherin, vimentin, β-catenin, and E-cadherin protein expressions. Compared with the blank group, the half maximal inhibitory concentration (IC50), number of cell colonies, number of invading cells and expressions of proteins related to EMT (i.e. N-cadherin, vimentin, and β-catenin) significantly reduced, but increases in apoptosis rate, p-p38 MAPK/p38 MAPK ratio and E-cadherin protein expression were observed in different groups (P < 0.05). Compared with the Platycodon grandiflorus + Curcumae Rhizoma group, the Platycodon grandiflorus + Curcumae Rhizoma + U46619 group had significantly decreased IC50, cell colony count, invading cell count and β-catenin, N-cadherin, and vimentin expressions, in addition to elevated E-cadherin protein expression, apoptosis rate, and p-p38 MAPK/p38 MAPK ratio (P < 0.05). Platycodon grandiflorus can reverse the resistance of breast cancer cells to Dox and regulate their biological activities by activating the p38 MAPK signaling pathway. Highlights: p38 MAPK expression enhances Dox resistance of breast cancer cells, regulates their biological activity and blocks their epithelial-mesenchymal transition. Platycodon grandiflorus can activate the p38 MAPK signaling pathway to reverse the resistance of breast cancer cells to Dox and regulate their biological activity. [ABSTRACT FROM AUTHOR]

This study investigated the utilization of by-products from Platycodon grandiflorus and the role of genetic diversity in their anti-melanogenic properties, focusing on the purple-flower (PF) and white-flower (WF) varieties. Our results show that the WF variety exhibited significantly higher anti-melanogenic activity, attributed to higher concentrations of key saponins such as platycodin D3 and platycodin D. These saponins demonstrated strong tyrosinase inhibitory effects as confirmed by molecular docking analysis. Further, the WF variety showed increased expression of genes involved in saponin biosynthesis, highlighting the role of genetic diversity in determining phytochemical composition and pharmacological efficacy. The superior anti-melanogenic activity of WF suggests its potential as a valuable ingredient in the cosmetic industry for skin-whitening products. Our findings emphasize the importance of utilizing by-products and selecting specific genotypes to enhance the quality and efficacy of plant-derived products. Future research should explore the full spectrum of bioactive compounds in P. grandiflorus, investigate sustainable extraction methods, and conduct clinical trials to validate the safety and effectiveness of these compounds in cosmetic and therapeutic applications. [ABSTRACT FROM AUTHOR]

Introduction: The uridine diphosphate (UDP)-glycosyltransferase (UGT) family is the largest glycosyltransferase family, which is involved in the biosynthesis of natural plant products and response to abiotic stress. UGT has been studied in many medicinal plants, but there are few reports on Platycodon grandiflorus. This study is devoted to genome-wide analysis of UGT family and identification of UGT genes involved in drought stress of Platycodon grandiflorus (PgUGTs). Methods: The genome data of Platycodon grandiflorus was used for genome-wide identification of PgUGTs, online website and bioinformatics analysis software was used to conduct bioinformatics analysis of PgUGT genes and the genes highly responsive to drought stress were screened out by qRT-PCR, these genes were cloned and conducted bioinformatics analysis. Results: A total of 75 PgUGT genes were identified in P.grandiflorus genome and clustered into 14 subgroups. The PgUGTs were distributed on nine chromosomes, containing multiple cis-acting elements and 22 pairs of duplicate genes were identified. Protein-protein interaction analysis was performed to predict the interaction between PgUGT proteins. Additionally, six genes were upregulated after 3d under drought stress and three genes (PGrchr09G0563, PGrchr06G0523, PGrchr06G1266) responded significantly to drought stress, as confirmed by qRT-PCR. This was especially true for PGrchr06G1266, the expression of which increased 16.21-fold after 3d of treatment. We cloned and conducted bioinformatics analysis of three candidate genes, both of which contained conserved motifs and several cis-acting elements related to stress response, PGrchr06G1266 contained the most elements. Discussion: PgGT1 was confirmed to catalyze the C-3 position of platycodin D and only eight amino acids showed differences between gene PGr008G1527 and PgGT1, which means PGr008G1527 may be able to catalyze the C-3 position of platycodin D in the same manner as PgGT1. Seven genes were highly expressed in the roots, stems, and leaves, these genes may play important roles in the development of the roots, stems, and leaves of P. grandiflorus. Three genes were highly responsive to drought stress, among which the expression of PGrchr06G1266 was increased 16.21-fold after 3d of drought stress treatment, indicating that PGrchr06G1266 plays an important role in drought stress tolerance. To summarize, this study laied the foundation to better understand the molecular bases of responses to drought stress and the biosynthesis of platycodin. [ABSTRACT FROM AUTHOR]

Abstract [Objective] Drought stress seriously affects the yield and quality of medicinal plants. This study aims to reveal the physiological mechanism of exogenous calcium in alleviating drought-induced damage in Platycodon grandifloras, which can provide a basis for using exogenous calcium to improve drought resistance and medicinal quality of P . grandiflorus in arid and semi-arid area. [Methods] With P . grandiflorus seedlings as materials, a pot experiment was conducted to explore the effects of exogenous calcium on the growth, photosynthetic gas exchange parameters, antioxidant enzyme activity, and medicinal quality of P . grandiflorus seedlings under drought stress by foliar spraying of 10 mmol/L CaCl2. [Results] (1) Exogenous calcium significantly increased root length and dry and fresh biomass of P . grandiflorus under drought stress. It significantly increased the stomatal aperture, chlorophyll a content, total chlorophyll content, carotenoid content, net photosynthetic rate, stomatal conductance, and transpiration rate by 30.28%, 57.67%, 44.44%, 100.33%, 89.53%, 60.00%, and 83.11%, respectively, in P . grandiflorus leaves under drought stress. (2) Exogenous calcium effectively reduced the content of malondialdehyde and hydrogen peroxide in P . grandiflorus leaves under drought stress, which was significantly reduced by 13.82% and 18.66%, respectively, compared to drought stress. Exogenous calcium increased the superoxide dismutase, superoxide dismutase, and catalase activities in P . grandiflorus leaves by 25.43%, 7.90%, and 33.92%, respectively, and also increased the soluble protein content in roots compared with drought stress. (3) Exogenous calcium significantly increased the levels of latycodin-D, polysaccharides, total flavonoids, and free amino acids in P . grandiflorus roots by 35.34%, 34.87%, 4.19%, and 6.52%, respectively, under normal conditions. Under drought stress, the levels were significantly increased by 10.94%, 7.53%, 6.07%, and 5.78%, respectively. (4) N, P, K, Ca, Cu, and Mn contents in the aboveground and underground parts of P . grandiflorus were significantly reduced under drought stress, while they were increased to varying degrees under exogenous calcium treatment. [Conclusion] Spraying 10 mmol/L CaCl2 could enhance the antioxidant system activity and osmotic regulation ability in P . grandiflorus leaves in arid environment, increase the content of photosynthetic pigments and photosynthetic performance, synergistically promote the accumulation of secondary metabolites and mineral elements, improve the quality of medicinal parts, and alleviate the damage of P . grandiflorus seedlings under drought stress.

Background: Platycodon grandiflorus belongs to the genus Platycodon and has many pharmacological effects, such as expectorant, antitussive, and anti-tumor properties. Among transcription factor families peculiar to eukaryotes, the basic leucine zipper (bZIP) family is one of the most important, which exists widely in plants and participates in many biological processes, such as plant growth, development, and stress responses. However, genomic analysis of the bZIP gene family and related stress response genes has not yet been reported in P. grandiflorus. Methods: P. grandiflorus bZIP (PgbZIP) genes were first identified here, and the phylogenetic relationships and conserved motifs in the PgbZIPs were also performed. Meanwhile, gene structures, conserved domains, and the possible protein subcellular localizations of these PgbZIPs were characterized. Most importantly, the cis-regulatory elements and expression patterns of selected genes exposed to two different stresses were analyzed to provide further information on PgbZIPs potential biological roles in P. grandiflorus upon exposure to environmental stresses. Conclusions: Forty-six PgbZIPs were identified in P. grandiflorus and divided into nine groups, as displayed in the phylogenetic tree. The results of the chromosomal location and the collinearity analysis showed that forty-six PgbZIP genes were distributed on eight chromosomes, with one tandem duplication event and eleven segmental duplication events identified. Most PgbZIPs in the same phylogenetic group have similar conserved motifs, domains, and gene structures. There are cis-regulatory elements related to the methyl jasmonate (MeJA) response, low-temperature response, abscisic acid response, auxin response, and gibberellin response. Ten PgbZIP genes were selected to study their expression patterns upon exposure to low-temperature and MeJA treatments, and all ten genes responded to these stresses. The real-time quantitative polymerase chain reaction (RT-qPCR) results suggest that the expression levels of most PgbZIPs decreased significantly within 6 h and then gradually increased to normal or above normal levels over the 90 h following MeJA treatment. The expression levels of all PgbZIPs were significantly reduced after 3 h of the low-temperature treatment. These results reveal the characteristics of the PgbZIP family genes and provide valuable information for improving P. grandiflorus's ability to cope with environmental stresses during growth and development. [ABSTRACT FROM AUTHOR]

The Basic Leucine Zipper (bZIP) transcription factors (TFs) family is among of the largest and most diverse gene families found in plant species, and members of the bZIP TFs family perform important functions in plant developmental processes and stress response. To date, bZIP genes in Platycodon grandiflorus have not been characterized. In this work, a number of 47 PgbZIP genes were identified from the genome of P. grandiflorus, divided into 11 subfamilies. The distribution of these PgbZIP genes on the chromosome and gene replication events were analyzed. The motif, gene structure, cis-elements, and collinearity relationships of the PgbZIP genes were simultaneously analyzed. In addition, gene expression pattern analysis identified ten candidate genes involved in the developmental process of different tissue parts of P. grandiflorus. Among them, Four genes (PgbZIP5, PgbZIP21, PgbZIP25 and PgbZIP28) responded to drought and salt stress, which may have potential biological roles in P. grandiflorus development under salt and drought stress. Four hub genes (PgbZIP13, PgbZIP30, PgbZIP32 and PgbZIP45) mined in correlation network analysis, suggesting that these PgbZIP genes may form a regulatory network with other transcription factors to participate in regulating the growth and development of P. grandiflorus. This study provides new insights regarding the understanding of the comprehensive characterization of the PgbZIP TFs for further exploration of the functions of growth and developmental regulation in P. grandiflorus and the mechanisms for coping with abiotic stress response.

Abstract Background Platycodon grandiflorus (Jacq.) A. DC is a famous traditional Chinese medicine in China and an authentic medicine in Inner Mongolia. It has been traditionally used as an expectorant in cough and also has anti-inflammatory and other pharmacological effects. As a homologous plant of medicine and food, P. grandiflorus is widely planted in Northeast China. Soil salinity isa limiting factor for its cultivation. In this study, we comprehensively described the physiological characteristics of P. grandiflorus and combined transcriptomics and metabolomics to study the response of roots of P. grandiflorus to salt stress. Results Overall, 8,988 differentially expressed genes were activated and significantly altered the metabolic processes. In total, 428 differentially abundant metabolites were affected by salt stress. After moderate and severe salt stress, most of the differentially abundant metabolites were enriched in the L-phenylalanine metabolic pathway. Through the comprehensive analysis of the interaction between key genes and metabolites, the main pathways such as lignin compound biosynthesis and triterpene saponin biosynthesis were completed. The relative content of compounds related to lignin biosynthesis, such as caffeic acid, coniferin, and syringing, increased under salt stress, and the related genes such as PAL, C4H, and the key enzyme gene UGT72E2 were activated to adapt to the salt stress. Platycodon saponin is one of the major triterpene saponins in P. grandiflorus, and Platycodin D is its most abundant major bioactive component. Under severe salt stress, Platycodin D level increased by nearly 1.77-fold compared with the control group. Most of the genes involved insynthetic pathway of Platycodin D, such as HMGCR, GGPS, SE, and LUP, were upregulated under salt stress. Conclusion Salt stress led to a decrease in the biomass and affected the activities of antioxidant enzymes and contents of osmotic regulators in the plant. These results provided not only novel insights into the underlying mechanisms of response of P. grandiflorus to salt stress but also a foundation for future studies on the function of genes related to salt tolerance in the triterpenoid saponin biosynthesis pathway.

This study investigated the utilization of by-products from Platycodon grandiflorus and the role of genetic diversity in their anti-melanogenic properties, focusing on the purple-flower (PF) and white-flower (WF) varieties. Our results show that the WF variety exhibited significantly higher anti-melanogenic activity, attributed to higher concentrations of key saponins such as platycodin D3 and platycodin D. These saponins demonstrated strong tyrosinase inhibitory effects as confirmed by molecular docking analysis. Further, the WF variety showed increased expression of genes involved in saponin biosynthesis, highlighting the role of genetic diversity in determining phytochemical composition and pharmacological efficacy. The superior anti-melanogenic activity of WF suggests its potential as a valuable ingredient in the cosmetic industry for skin-whitening products. Our findings emphasize the importance of utilizing by-products and selecting specific genotypes to enhance the quality and efficacy of plant-derived products. Future research should explore the full spectrum of bioactive compounds in P. grandiflorus, investigate sustainable extraction methods, and conduct clinical trials to validate the safety and effectiveness of these compounds in cosmetic and therapeutic applications.

The consumption of agricultural water resources is enormous, and in the face of water scarcity, optimizing the utilization rate of water resources is an urgent need. As a common medicinal plant, the planting area of Platycodon grandiflorus crops has increased. Therefore, in order to maximize the yield and quality of Platycodon crops, different irrigation treatment methods are developed in this study based on their evaporation capacity and the impact of different irrigation amounts on Platycodon crops is explored. Different irrigation amounts affect various indicators of Platycodon plants. The experimental results of this study indicate that for the daily transpiration of Platycodon, 120% irrigation is more effective during the maturity period, and the stemflow efficiency performs better overall on sunny days. Irrigation at a rate of 120% can obtain the maximum number of leaves. Regarding the quality of Platycodon grandiflorus crops, 120% irrigation can achieve the highest contents of saponins, flavonoids, and polysaccharides, but 100% irrigation is required to achieve the highest soluble protein contents. Overall, Platycodon crops require 100% irrigation during the flowering stage and 120% irrigation during the maturity stage. As the temperature increases, the irrigation amount should be appropriately increased, and it should be reduced as the temperature decreases. This research demonstrates the impact of irrigation volume on Platycodon crops, and controlling the irrigation volume can effectively enhance the utilization efficiency of water resources. [ABSTRACT FROM AUTHOR]

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Background: Platycodon grandiflorus (Jacq.) A. DC is a famous traditional Chinese medicine in China and an authentic medicine in Inner Mongolia. It has been traditionally used as an expectorant in cough and also has anti-inflammatory and other pharmacological effects. As a homologous plant of medicine and food, P. grandiflorus is widely planted in Northeast China. Soil salinity isa limiting factor for its cultivation. In this study, we comprehensively described the physiological characteristics of P. grandiflorus and combined transcriptomics and metabolomics to study the response of roots of P. grandiflorus to salt stress. Results: Overall, 8,988 differentially expressed genes were activated and significantly altered the metabolic processes. In total, 428 differentially abundant metabolites were affected by salt stress. After moderate and severe salt stress, most of the differentially abundant metabolites were enriched in the L-phenylalanine metabolic pathway. Through the comprehensive analysis of the interaction between key genes and metabolites, the main pathways such as lignin compound biosynthesis and triterpene saponin biosynthesis were completed. The relative content of compounds related to lignin biosynthesis, such as caffeic acid, coniferin, and syringing, increased under salt stress, and the related genes such as PAL, C4H, and the key enzyme gene UGT72E2 were activated to adapt to the salt stress. Platycodon saponin is one of the major triterpene saponins in P. grandiflorus, and Platycodin D is its most abundant major bioactive component. Under severe salt stress, Platycodin D level increased by nearly 1.77-fold compared with the control group. Most of the genes involved insynthetic pathway of Platycodin D, such as HMGCR, GGPS, SE, and LUP, were upregulated under salt stress. Conclusion: Salt stress led to a decrease in the biomass and affected the activities of antioxidant enzymes and contents of osmotic regulators in the plant. These results provided not only novel insights into the underlying mechanisms of response of P. grandiflorus to salt stress but also a foundation for future studies on the function of genes related to salt tolerance in the triterpenoid saponin biosynthesis pathway. [ABSTRACT FROM AUTHOR]

Platycodon grandiflorus set ornamental, edible, and medicinal plant with broad prospects for further application development. However, there are no reports on the YABBY transcription factor in P. grandiflorus. Identification and analysis of the YABBY gene family of P. grandiflorus using bioinformatics means. Six YABBY genes were identified and divided into five subgroups. Transcriptome data and qRT-PCR were used to analyze the expression patterns of YABBY. YABBY genes exhibited organ-specific patterns in expression in P grandiflorus. Upon salt stress and drought induction, P. grandiflorus presented different morphological and physiological changes with some dynamic changes. Under salt treatment, the YABBY gene family was down-regulated; PgYABBY5 was up-regulated in leaves at 24 h. In drought treatment, PgYABBY1, PgYABBY2, and PgYABBY3 were down-regulated to varying degrees, but PgYABBY3 was significantly up-regulated in the roots. PgYABBY5 was up-regulated gradually after being down-regulated. PgYABBY5 was significantly up-regulated in stem and leaf at 48 h. PgYABBY6 was down-regulated at first and then significantly up-regulated. The dynamic changes of salt stress and drought stress can be regarded as the responses of plants to resist damage. During the whole process of salt and drought stress treatment, the protein content of each tissue part of P grandiflorus changed continuously. At the same time, we found that the promoter region of the PgYABBY gene contains stress-resistant elements, and the regulatory role of YABBY transcription factor in the anti-stress mechanism of P grandiflorus remains to be studied. PgYABBY1, PgYABBY2, and PgYABBY5 may be involved in the regulation of saponins in P. grandiflorus. PgYABBY5 may be involved in the drought resistance mechanism in P. grandiflorus stems and leaves. This study may provide a theoretical basis for studying the regulation of terpenoids by the YABBY transcription factor and its resistance to abiotic stress.

Objective: To investigate the impact of developmental stages on triterpene saponin content and expression of key genes in their biosynthetic pathway in Platycodon grandiflorus seedlings. Method: The total saponins were extracted using ultrasonic extraction with methanol as the solvent. The quantification of Platycodin D in 6-month-old and 1-year-old Platycodon grandiflorus seedlings was performed using ultra-performance liquid chromatography tandem mass spectrometry, while the expression of key genes in the biosynthesis of saponins was determined by real-time quantitative polymerase chain reaction. Result: The content of Platycodin D and total saponins in 1-year-old Platycodon grandiflorus roots was significantly higher than that in of 6-month-old. The expression of key genes in saponin biosynthesis, such as PGMVK, PGMVD, PGDXR, PGMCT, PGMDS, PGHDS, PGHDR, PGFPS, and PGSS, was higher in 1-year-old roots compared to 6-month-old seedlings. Conclusion: The medicinal value of 1-year-old Platycodon grandiflorus root was higher than that of 6-month-old seedlings. The difference in medicinal value can be attributed to the variation in the expression of key genes involved in the saponin biosynthesis pathway. [ABSTRACT FROM AUTHOR]

While the status of histone acetylation is a critical regulator of chromatin's structure with a significant impact on plant physiology, our understanding of epigenetic regulation in the biosynthesis of active compounds in plants is limited. In this study, Platycodon grandiflorus was treated with sodium butyrate (NaB), a histone deacetylase inhibitor, to investigate the influence of histone acetylation on secondary metabolism. Its treatment with NaB increased the acetylation of histone H3 at lysine 9, 14, and 27 and enhanced the anti-melanogenic properties of P. grandiflorus roots. Through transcriptome and differentially expressed gene analyses, we found that NaB influenced the expression of genes that were involved in both primary and secondary metabolic pathways. In addition, NaB treatment caused the accumulation of polyphenolic compounds, including dihydroquercetin, gallic acid, and 2,4-dihydroxybenzoic acid. The NaB-induced transcriptional activation of genes in the phenylpropanoid biosynthetic pathway influenced the anti-melanogenic properties of P. grandiflorus roots. Overall, these findings suggest the potential of an epigenomic approach to enhance the medicinal qualities of medicinal plants. [ABSTRACT FROM AUTHOR]

Liang Wang,1,&ast; Longjiao Hu,2,&ast; Zhilian Peng,3 Honghong Cao,1 Danfeng Cao,4 Yiqin Long,4 Zhengyu Zou4 1Pharmacy Department, Traditional Chinese Medicine Hospital of Jiulongpo Distriction Chongqing, Chongqing, People’s Republic of China; 2Pharmacy Department, Chongqing Jiulongpo People’s Hospital, Chongqing, People’s Republic of China; 3Administrative Department, Traditional Chinese Medicine Hospital of Jiulongpo Distriction Chongqing, Chongqing, People’s Republic of China; 4Clinical Laboratory, Chongqing Jiulongpo People’s Hospital, Chongqing, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Zhengyu Zou, Clinical Laboratory, Chongqing Jiulongpo People’s Hospital, Chongqing, People’s Republic of China, Email hulongjiao494@126.com Liang Wang, Pharmacy Department, Traditional Chinese Medicine Hospital of Jiulongpo Distriction Chongqing, Chongqing, People’s Republic of China, Email askliang621129@126.comBackground: Platycodon grandiflorus could significantly improve the pathological results of cutaneous scald injury, reduce the release of inflammatory factors and promote angiogenesis. This study investigated the wound healing effect of luteolin, an active component of P. grandiflorus, on induced cutaneous scald injury in Sprague-Dawley (SD) rats.Methods: The protein expression levels of TNF-α and IL-6 were detected by ELISA. QRT-PCR was adopted to detect the expression of TGF-β 1 and VEGF. Histopathological changes of scald wounds were analyzed by hematoxylin-eosin staining. Cell viability and migration ability were detected by CCK-8 assay and scratch assay.Results: Both in vivo and in vitro experiments showed that luteolin promoted wound healing of cutaneous scald injury. Gene Oncology (GO) functional analysis and rescue experiments showed that endothelial nitric oxide synthase 3 (NOS3) was the critical target of luteolin in treating cutaneous scald.Conclusion: This study demonstrated that luteolin is an effective component of P. grandiflorus and is effective in the treatment of cutaneous scald injury.Keywords: Platycodon grandiflorus, luteolin, scald injury, nitric oxide synthase, network pharmacology

Platycodon grandiflorus (balloon flower), used as a food reserve as well as in traditional herbal medicine, is known for its multiple beneficial effects. In particular, this plant is widely used as a vegetable in Republic of Korea. We examined the ameliorative effects of P. grandiflorus on alloxan-induced pancreatic islet damage in zebrafish. The aerial part treatment led to a significant recovery in pancreatic islet size and glucose uptake. The efficacy of the aerial part was more potent than that of the root. Eight flavonoids (1–8) were isolated from the aerial part. Structures of two new flavone glycosides, designated dorajiside I (1) and II (2), were elucidated to be luteolin 7-O-α-L-rhamno-pyranosyl (1 → 2)-(6-O-acetyl)-β-D-glucopyranoside and apigenin 7-O-α-L-rhamnopyranosyl (1 → 2)-(6-O-acetyl)-β-D-glucopyranoside, respectively, by spectroscopic analysis. Compounds 1, 3, 4 and 6–8 yielded the recovery of injured pancreatic islets in zebrafish. Among them, compound 7 blocked KATP channels in pancreatic β-cells. Furthermore, compounds 3, 4, 6 and 7 showed significant changes with respect to the mRNA expression of GCK, GCKR, GLIS3 and CDKN2B compared to alloxan-induced zebrafish. In conclusion, the aerial part of P. grandiflorus and its constituents conferred a regenerative effect on injured pancreatic islets. [ABSTRACT FROM AUTHOR]

A novel β-carboline, namely Plagranline A (1), and a known alkaloid (2) were isolated from the roots of Platycodon grandiflorus. Their structure was elucidated using a combination of spectroscopic analyses and computational chemistry, including electronic circular dichroism (ECD) and optical rotatory dispersion (ORD). Alkaloids 1 and 2 exhibited moderate inhibitory effects against NO production. Interestingly, 1 and 2 are the first alkaloid constituents reported from the plant P. grandifloras. [ABSTRACT FROM AUTHOR]

Floral color plays a major role in pollinator specificity, and changes in color may result in pollinator shifts and pollinator-mediated speciation. In the purple flowers of Platycodon grandiflorus, anthocyanins are the major pigment metabolites, whereas white flowers result due to the absence of anthocyanins. The lack of anthocyanins may be due to the inhibition of the anthocyanin biosynthesis pathway. However, the molecular mechanism of anthocyanin biosynthesis in P. grandiflorus is not fully understood. Hence, we identified R2R3-MYB transcription factor, PlgMYBR1, as a negative regulator for anthocyanin biosynthesis using sequence homology and tissue-specific expression pattern analyses. A heterologous co-expression assay suggested that PlgMYBR1 inhibited the function of AtPAP1 (Arabidopsis thaliana production of anthocyanin pigment 1), indicating that PlgMYBR1 plays as a repressor of anthocyanin biosynthesis in P. grandiflorus. Our results provide a foundation for future efforts to understand the anthocyanin biosynthesis in P. grandiflorus and, thereby, to improve flower color through genetic engineering. [ABSTRACT FROM AUTHOR]

Platycodon grandiflorus, a perennial flowering plant widely distributed in China and South Korea, is an excellent resource for both food and medicine. The main active compounds of P. grandiflorus are triterpenoid saponins. WRKY transcription factors (TFs) are among the largest gene families in plants and play an important role in regulating plant terpenoid accumulation, physiological metabolism, and stress response. Numerous studies have been reported on other medicinal plants; however, little is known about WRKY genes in P. grandiflorus. In this study, 27 PgWRKYs were identified in the P. grandiflorus transcriptome. Phylogenetic analysis showed that PgWRKY genes were clustered into three main groups and five subgroups. Transcriptome analysis showed that the PgWRKY gene expression patterns in different tissues differed between those in Tongcheng City (Southern Anhui) and Taihe County (Northern Anhui). Gene expression analysis based on RNA sequencing and qRT-PCR analysis showed that most PgWRKY genes were expressed after induction with methyl jasmonate (MeJA). Co-expressing PgWRKY genes with triterpenoid biosynthesis pathway genes revealed four PgWRKY genes that may have functions in triterpenoid biosynthesis. Additionally, functional annotation and protein-protein interaction analysis of PgWRKY proteins were performed to predict their roles in potential regulatory networks. Thus, we systematically analyzed the structure, evolution, and expression patterns of PgWRKY genes to provide an important theoretical basis for further exploring the molecular basis and regulatory mechanism of WRKY TFs in triterpenoid biosynthesis.

Platycodin D (PD) is a deglycosylated triterpene saponin with much higher pharmacological activity than glycosylated platycoside E (PE). Extensive studies in vitro showed that the transformation of platycoside E to platycodin D can be achieved using ß-glucosidase extracted from several bacteria. However, whether similar enzymes in Platycodon grandiflorus could convert platycoside E to platycodin D, as well as the molecular mechanism underlying the deglycosylation process of platycodon E, remain unclear. Here, we identified a ß-glucosidase in P. grandiflorus from our previous RNA-seq analysis, with a full-length cDNA of 1,488 bp encoding 495 amino acids. Bioinformatics and phylogenetic analyses showed that ß-glucosidases in P. grandiflorus have high homology with other plant ß-glucosidases. Subcellular localization showed that there is no subcellular preference for its encoding gene. ß-glucosidase was successfully expressed as 6 × His-tagged fusion protein in Escherichia coli BL21 (DE3). Western blot analysis yielded a recombinant protein of approximately 68 kDa. In vitro enzymatic reactions determined that ß-glucosidase was functional and could convert PE to PD. RT-qPCR analysis showed that the expression level of ß-glucosidase was higher at night than during the day, with the highest expression level between 9:00 and 12:00 at night. Analysis of the promoter sequence showed many light-responsive cis-acting elements, suggesting that the light might regulate the gene. The results will contribute to the further study of the biosynthesis and metabolism regulation of triterpenoid saponins in P. grandiflorus. [ABSTRACT FROM AUTHOR]

Platycodon grandiflorus is a medical, ornamental, and edible traditional Chinese medicine whose seed germination and plant growth are frequently restricted by dormancy and stresses. In this study, we investigated how chitosan soaking affected seed germination, growth, photosynthesis, resistance, yield, and quality of P. grandiflorus. The results indicated that chitosan soaking had a preferable enhancing effect on seed germination of P. grandiflorus, which significantly (p < 0.05) promoted its germination rate, energy, and index, as well as cotyl and radicle length. Furthermore, 0.15–0.20% chitosan soaking effectively enhanced the leaf growth, height, stem diameter, and overground part dry weight of P. grandiflorus and reliably improved their leaves’ chlorophyll, photosynthetic rate, transpiration rate, and water use efficiency. Moreover, 0.15–0.20% chitosan soaking effectively enhanced the stress resistance and adaptability of P. grandiflorus via increasing its resistance substances and triggering its defense enzyme activity. Meanwhile, 0.15–0.20% chitosan soaking effectively improved the underground part growth and medical quality of P. grandiflorus. This study highlights that chitosan can be used as a favorable, efficient, and economical candidate or promoter for enhancing seed germination of P. grandiflorus and improving its growth, photosynthesis, resistance, yield, and quality; it also highlights that 0.15–0.20% chitosan is a suitable concentration.

The Basic Leucine Zipper (bZIP) transcription factors (TFs) family is among of the largest and most diverse gene families found in plant species, and members of the bZIP TFs family perform important functions in plant developmental processes and stress response. To date, bZIP genes in Platycodon grandiflorus have not been characterized. In this work, a number of 47 PgbZIP genes were identified from the genome of P. grandiflorus , divided into 11 subfamilies. The distribution of these PgbZIP genes on the chromosome and gene replication events were analyzed. The motif, gene structure, cis -elements, and collinearity relationships of the PgbZIP genes were simultaneously analyzed. In addition, gene expression pattern analysis identified ten candidate genes involved in the developmental process of different tissue parts of P. grandiflorus . Among them, Four genes ( PgbZIP5 , PgbZIP21 , PgbZIP25 and PgbZIP28 ) responded to drought and salt stress, which may have potential biological roles in P. grandiflorus development under salt and drought stress. Four hub genes ( PgbZIP13 , PgbZIP30 , PgbZIP32 and PgbZIP45 ) mined in correlation network analysis, suggesting that these PgbZIP genes may form a regulatory network with other transcription factors to participate in regulating the growth and development of P. grandiflorus . This study provides new insights regarding the understanding of the comprehensive characterization of the PgbZIP TFs for further exploration of the functions of growth and developmental regulation in P. grandiflorus and the mechanisms for coping with abiotic stress response., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Wang, Wang, Cao, Liu, Kong, Wang, Ren, Fu and Ma.)

Featured Application: Authors are encouraged to provide a concise description of the specific application or a potential application of the work. This section is not mandatory.Platycodon grandiflorus has huge potential medicinal applications in different diseases and immune enhancement (IE). In the past, limited studies have been conducted to decipher the molecular mechanism behind IE of Platycodon grandiflorus extract (PGE) despite the fact that IE can be one of the important factors for application of PGE in different diseases including cancer. In this study, whole transcriptome expression and subsequent analysis in PGE treated/untreated organisms were carried out to explore the molecular mechanism behind IE, and subsequently anticancer effect. Current study has the possibility of interaction of PGE treatment with diet, so further study is required to validate the results. The immunity-related genes were found to be up-regulated in the differential expression analysis, which is in line with the IE potential of PGE and the literature. Similarly, Siglec-15, the immune-suppressor gene that has recently been suggested as the normalization target in cancer, was one of the most down-regulated genes. In a set analysis with immune-suppressor genes, the high number of immune-suppressor genes were found to be down-regulated, which indicated that down-regulation of immune-suppressor genes can be the major mechanism behind the IE in PGE treatment. Identified genes, important in immunity and cancer, are highly recommended for anticancer and IE targets in future studies. [ABSTRACT FROM AUTHOR]

Background: The root of Platycodon grandiflorus (PG) has a long-standing tradition in the Asian diet and herbal medicine, because of its anti-inflammatory and antiobesity effects. Changes in the gut microbiota can have dietary effects on host health, which suggests a relation between the 2.Objectives: The aim of our study was to investigate the relation between PG-mediated suppression of obesity and the composition and functioning of the gut microbiota.Methods: Six-week-old male C57BL/6J mice were fed either a control diet (CON, 10% kcal from fat), a high-fat diet (HFD, 60% kcal from fat), or a PG-supplemented HFD for 18 wk. PG was administered by oral gavage at 2 g · kg body weight-1 · d-1. Body weight and food intake were monitored. Lipid metabolism, inflammation, and intestinal barrier function were determined. Amplicon sequencing of the bacterial 16S ribosomal RNA gene was used to explore gut microbiota structure, and nontargeted metabolomics analysis was performed to investigate metabolite concentrations in fecal samples.Results: We found that PG significantly ameliorated HFD-induced inflammation, recovered intestinal barrier integrity (reduced permeability by 39% , P = 0.008), reduced fat accumulation by 26% (P = 0.009), and changed the expression of key genes involved in the development of white adipose tissue (P < 0.05) in HFD-fed mice to similar levels in CON mice. Moreover, PG attenuated HFD-induced changes in the gut microbiota; it especially increased Allobaculum (7.3-fold, P = 0.002) relative to HFD, whereas CON was 15.2-fold of HFD (P = 0.002). These changes by PG were associated with an increase in the production of SCFAs (butyrate and propionate, P < 0.001) and other carbohydrate-related metabolites known to have a major role in disease suppression.Conclusions: Our study demonstrated that PG beneficially changed the gut microbiota and the gut metabolome in HFD-fed mice, and suggests that the antiobesity effects of PG may be mediated via changes in gut microbiota composition and metabolic activity. [ABSTRACT FROM AUTHOR]

Platycodon grandiflorus roots have been used as a foodstuff and traditional medicine for thousands of years in East Asia. In order to increase the root development of P. grandiflorus, cultivators removed the inflorescences, suggesting the possible negative effect of flowering on root development. This indicates that the genetic improvement of P. grandiflorus by late flowering is a potential approach to increase productivity. However, nothing is known about key genes integrating multiple flowering pathways in P. grandiflorus. In order to fill this gap, we identified potential homologs of the FLOWERING LOCUS T (FT) gene in P. grandiflorus. The alignment with other FT members and phylogenetic analysis revealed that the P. grandiflorus FT (PlgFT) protein contains highly conserved functional domains and belongs to the FT-like clade. The expression analysis revealed spatial variations in the transcription of PlgFT in different organs. In addition, the expression level of PlgFT was increased by high temperature but not by photoperiodic light input signals, presumably due to lacking the CONSTANS binding motif in its promoter region. Furthermore, PlgFT induced early flowering upon its overexpression in P. grandiflorus, suggesting the functional role of PlgFT in flowering. Taken together, we functionally characterized PlgFT as a master regulator of P. grandiflorus flowering under inductive high temperature, which will serve as an important target gene for improving the root productivity.

Platycodon grandiflorus is the single genus plant in family Campanulaceae, which is distributed in areas of China, Japan, Korea, and Russia. In this study, we assembled and annotated the complete chloroplast genome of P. grandiflorus. The chloroplast genome of P. grandiflorus is 171,624 bp long, containing a shorter small single-copy region (SSC) of 7,962 bp, a normal large single-copy region (LSC) of 79,690 bp and a longer pair of inverted repeat regions (IRs) of 41,986 bp. The whole chloroplast genome of P. grandiflorus has 146 genes, including 102 protein-coding genes (PCGs), 36 transfer RNA genes (tRNAs), and 8 ribosome RNA genes (rRNAs). The overall nucleotide composition is: A of 30.9%, T of 30.9%, C of 19.3%, and G of 18.9%, with a total G + C content of the chloroplast genome 38.2%. The phylogenetic and genetic analysis showed a close relationship between Platycodon grandiflorus and Codonopsis lanceolata.

Background: Platycodon grandiflorus belongs to the genus Platycodon and has many pharmacological effects, such as expectorant, antitussive, and anti-tumor properties. Among transcription factor families peculiar to eukaryotes, the basic leucine zipper (bZIP) family is one of the most important, which exists widely in plants and participates in many biological processes, such as plant growth, development, and stress responses. However, genomic analysis of the bZIP gene family and related stress response genes has not yet been reported in P. grandiflorus ., Methods: P. grandiflorus bZIP (PgbZIP) genes were first identified here, and the phylogenetic relationships and conserved motifs in the PgbZIPs were also performed. Meanwhile, gene structures, conserved domains, and the possible protein subcellular localizations of these PgbZIPs were characterized. Most importantly, the cis-regulatory elements and expression patterns of selected genes exposed to two different stresses were analyzed to provide further information on PgbZIPs potential biological roles in P. grandiflorus upon exposure to environmental stresses., Conclusions: Forty-six PgbZIPs were identified in P. grandiflorus and divided into nine groups, as displayed in the phylogenetic tree. The results of the chromosomal location and the collinearity analysis showed that forty-six PgbZIP genes were distributed on eight chromosomes, with one tandem duplication event and eleven segmental duplication events identified. Most PgbZIPs in the same phylogenetic group have similar conserved motifs, domains, and gene structures. There are cis-regulatory elements related to the methyl jasmonate (MeJA) response, low-temperature response, abscisic acid response, auxin response, and gibberellin response. Ten PgbZIP genes were selected to study their expression patterns upon exposure to low-temperature and MeJA treatments, and all ten genes responded to these stresses. The real-time quantitative polymerase chain reaction (RT-qPCR) results suggest that the expression levels of most PgbZIPs decreased significantly within 6 h and then gradually increased to normal or above normal levels over the 90 h following MeJA treatment. The expression levels of all PgbZIPs were significantly reduced after 3 h of the low-temperature treatment. These results reveal the characteristics of the PgbZIP family genes and provide valuable information for improving P. grandiflorus's ability to cope with environmental stresses during growth and development., Competing Interests: The authors declare that they have no competing interests., (© 2024 Fan et al.)

Introduction: The uridine diphosphate (UDP)-glycosyltransferase (UGT) family is the largest glycosyltransferase family, which is involved in the biosynthesis of natural plant products and response to abiotic stress. UGT has been studied in many medicinal plants, but there are few reports on Platycodon grandiflorus. This study is devoted to genome-wide analysis of UGT family and identification of UGT genes involved in drought stress of Platycodon grandiflorus (PgUGTs)., Methods: The genome data of Platycodon grandiflorus was used for genome-wide identification of PgUGTs, online website and bioinformatics analysis software was used to conduct bioinformatics analysis of PgUGT genes and the genes highly responsive to drought stress were screened out by qRT-PCR, these genes were cloned and conducted bioinformatics analysis., Results: A total of 75 PgUGT genes were identified in P.grandiflorus genome and clustered into 14 subgroups. The PgUGTs were distributed on nine chromosomes, containing multiple cis-acting elements and 22 pairs of duplicate genes were identified. Protein-protein interaction analysis was performed to predict the interaction between PgUGT proteins. Additionally, six genes were upregulated after 3d under drought stress and three genes (PGrchr09G0563, PGrchr06G0523, PGrchr06G1266) responded significantly to drought stress, as confirmed by qRT-PCR. This was especially true for PGrchr06G1266, the expression of which increased 16.21-fold after 3d of treatment. We cloned and conducted bioinformatics analysis of three candidate genes, both of which contained conserved motifs and several cis-acting elements related to stress response, PGrchr06G1266 contained the most elements., Discussion: PgGT1 was confirmed to catalyze the C-3 position of platycodin D and only eight amino acids showed differences between gene PGr008G1527 and PgGT1, which means PGr008G1527 may be able to catalyze the C-3 position of platycodin D in the same manner as PgGT1. Seven genes were highly expressed in the roots, stems, and leaves, these genes may play important roles in the development of the roots, stems, and leaves of P. grandiflorus. Three genes were highly responsive to drought stress, among which the expression of PGrchr06G1266 was increased 16.21-fold after 3d of drought stress treatment, indicating that PGrchr06G1266 plays an important role in drought stress tolerance. To summarize, this study laied the foundation to better understand the molecular bases of responses to drought stress and the biosynthesis of platycodin., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Chen, Wang, Yu, Dong, Li, Chang, Wang, Jiang, Liu, Chi, Zha and Gui.)

Adipocytes regulate lipid metabolism according to physiological energy requirements. A dysfunctional lipid metabolism can lead to obesity and its complications such as hepatic steatosis, diabetes, and hyperlipidemia. In our study, the impact of Platycodon grandiflorus root ethanol extract (PGH) on lipid excretion and thermogenesis-related markers in diet-induced obesity mice was analyzed. Our data show that PGH elevated fatty acid uptake in epididymal adipose tissue by increasing Cd36, Slc27a1, Ffar2, and Ffar4 expression, which led to decreased blood free fatty acid concentrations. Moreover, PGH normalized body weight and fat mass in diet-induced obese mice by increasing lipolysis (Plin1, Atgl, and Hsl) and fatty acid oxidation. Changes in the levels of browning-related genes, enzyme activity of carnitine palmitoyltransferase, and the overall transcriptome (Bmp4, Cidec, Ucp3, Sirt3, and Cox4i1) led to promote brown adipose tissue-like features (browning) in epididymal white adipose tissue and enhanced energy expenditure. Our results suggest that PGH promotes lipid excretion and thermogenic function in high-fat diet-induced obese mice, which are mediated by regulation of fat metabolism. [ABSTRACT FROM AUTHOR]

Purpose: To evaluate the healing effect of Platycodon grandiflorus (Jacq.). extract (PGE) on experimental burn wounds in rats. Methods: Rats were randomly divided into four groups of eight rats each: control group, silver sulfadiazine (SSD)-treated group, moist exposed burn ointment (MEBO)-treated group and PGE-treated group. PGE, SSD and MEBO were applied topically twice daily for 7 days. SSD and MEBO were used as reference control. External observation of wound area contraction and histological analysis of wound tissues was performed respectively. The effect of PGE on matrix metalloproteases (MMPs), vascular endothelial growth factor (VEGF) and Type-III Collagen proteins of wound tissue in rats were analysed by Western blot. Results: After 10 days of topical treatment with PGE, PGE-treated group showed faster reduction in wound area when compared with control groups (p < 0.01). Matrix metalloprotease-2 (MMP-2), matrix metalloprotease-9 (MMP-9), VEGF and type-III collagen expressions in the wound tissue increased significantly (p < 0.05) when compared with the control burn wounds. Histological results showed an overall early recovery and regeneration in PGE-treated group when compared with control group. Conclusion: PGE possesses a significant wound-healing activity in full-thickness burn wounds in rats. Therefore, it can potentially be developed for the management of burns. [ABSTRACT FROM AUTHOR]

Platycodon grandiflorus has huge potential medicinal applications in different diseases and immune enhancement (IE). In the past, limited studies have been conducted to decipher the molecular mechanism behind IE of Platycodon grandiflorus extract (PGE) despite the fact that IE can be one of the important factors for application of PGE in different diseases including cancer. In this study, whole transcriptome expression and subsequent analysis in PGE treated/untreated organisms were carried out to explore the molecular mechanism behind IE, and subsequently anticancer effect. Current study has the possibility of interaction of PGE treatment with diet, so further study is required to validate the results. The immunity-related genes were found to be up-regulated in the differential expression analysis, which is in line with the IE potential of PGE and the literature. Similarly, Siglec-15, the immune-suppressor gene that has recently been suggested as the normalization target in cancer, was one of the most down-regulated genes. In a set analysis with immune-suppressor genes, the high number of immune-suppressor genes were found to be down-regulated, which indicated that down-regulation of immune-suppressor genes can be the major mechanism behind the IE in PGE treatment. Identified genes, important in immunity and cancer, are highly recommended for anticancer and IE targets in future studies.

Platycodon grandiflorus set ornamental, edible, and medicinal plant with broad prospects for further application development. However, there are no reports on the YABBY transcription factor in P. grandiflorus . Identification and analysis of the YABBY gene family of P. grandiflorus using bioinformatics means. Six YABBY genes were identified and divided into five subgroups. Transcriptome data and qRT-PCR were used to analyze the expression patterns of YABBY. YABBY genes exhibited organ-specific patterns in expression in P grandiflorus . Upon salt stress and drought induction, P. grandiflorus presented different morphological and physiological changes with some dynamic changes. Under salt treatment, the YABBY gene family was down-regulated; PgYABBY5 was up-regulated in leaves at 24 h. In drought treatment, PgYABBY1, PgYABBY2 , and PgYABBY3 were down-regulated to varying degrees, but PgYABBY3 was significantly up-regulated in the roots. PgYABBY5 was up-regulated gradually after being down-regulated. PgYABBY5 was significantly up-regulated in stem and leaf at 48 h. PgYABBY6 was down-regulated at first and then significantly up-regulated. The dynamic changes of salt stress and drought stress can be regarded as the responses of plants to resist damage. During the whole process of salt and drought stress treatment, the protein content of each tissue part of P grandiflorus changed continuously. At the same time, we found that the promoter region of the PgYABBY gene contains stress-resistant elements, and the regulatory role of YABBY transcription factor in the anti-stress mechanism of P grandiflorus remains to be studied. PgYABBY1, PgYABBY2 , and PgYABBY5 may be involved in the regulation of saponins in P. grandiflorus. PgYABBY5 may be involved in the drought resistance mechanism in P. grandiflorus stems and leaves. This study may provide a theoretical basis for studying the regulation of terpenoids by the YABBY transcription factor and its resistance to abiotic stress.

Platycodon grandiflorus is a widely used edible, traditional Chinese medicinal herb. It is rich in saponins, flavonoids, phenolic acids, and other compounds. It contains a large number of fatty acids such as linoleic acid (up to 63.24%), a variety of amino acids, vitamins, and multiple essential trace elements. P. grandiflorus has several biological applications, such as in hypotension, lipid reduction, atherosclerosis, inflammation, relieving cough and phlegm, promoting cholic acid secretion, and as an antioxidant. Further, P. grandiflorus is often used in the development of cold mixed vegetables, canned vegetables, preserved fruit, salted vegetables, and cosmetics in northeast China, South Korea, Japan, and Korea. In this paper, the active chemical components and the health benefits of P. grandiflorus have been reviewed, providing new ideas for the further development of nutraceutical products to prevent and manage chronic diseases.

Platycodon grandiflorus is a plant widely used in traditional Chinese medicine, of which polysaccharides are reported to be the main components responsible for its bio-functions. In this work, the inulin-type fructan (PGF) was obtained by DEAE anion exchange chromatography from the water extracted from P. grandifloras. Characterization was performed with methanolysis, methylation, and NMR and the results showed that PGF is a β-(2-1) linked fructan, with terminal glucose and with a degree of polymerization of 2–10. In order to study its biofunctions, the prebiotic and immunomodulation properties were assayed. We found that PGF exhibited good prebiotic activity, as shown by a promotion on six strains of lactobacillus proliferation. Additionally, the PGF also displayed direct immunomodulation on intestinal epithelial cells and stimulated the expressions of anti-inflammatory factors. These results indicated that the inulin from P. grandiflorus is a potential natural source of prebiotics as well as a potential intestinal immunomodulator, which will be valuable for further studies and new applications.

Platycodon grandiflorus (Jacq.) A.DC. (PG) has long been used as an ingredient of foods and is known to have beneficial effects on cognitive functions as well. The present study examined the effect of each PG extract (PGE) from root, aerial part, and seeds on cognitive functions in mice. Changes in spatial learning and memory using a Y-maze test, and markers of adult hippocampal neurogenesis and synaptogenesis were examined. Moreover, changes in neuritogenesis and activation of the ERK1/2 pathway were investigated. Results indicated that mice administered PGE (root) showed increased spontaneous alternation in the Y-maze test and synaptogenesis in the hippocampus. In addition, PGE (root) and platycodin D, the major bioactive compound from the PG root, significantly stimulated neuritic outgrowth by phosphorylation of the ERK1/2 signaling pathway in vitro. These results indicate that the PGE (root), containing platycodin D, enhances cognitive function through synaptogenesis via activation of the ERK1/2 signaling pathway. [ABSTRACT FROM AUTHOR]

The roots of Platycodon grandiflorus are widely used as a crude drug. The active components include a variety of triterpenoid saponins. Recent studies have revealed that Cyt P450 monooxygenases (P450s) function as triterpene oxidases in triterpenoid saponin biosynthesis in many plant species. However, there have been no reports regarding triterpene oxidases in P. grandiflorus. In this study, we performed transcriptome analysis of three different P. grandiflorus tissues (roots, leaves and petals) using RNA sequencing (RNA-Seq) technology. We cloned six P450 genes that were highly expressed in roots, and classified them as belonging to the CYP716A, CYP716D and CYP72A subfamilies. We heterologously expressed these P450s in an engineered yeast strain that produces β-amyrin, one of the most common triterpenes in plants. Two of the CYP716A subfamily P450s catalyzed oxidation reactions of the β-amyrin skeleton. One of these P450s, CYP716A140v2, catalyzed a three-step oxidation reaction at C-28 on β-amyrin to produce oleanolic acid, a reaction performed by CYP716A subfamily P450s in a variety of plant species. The other P450, CYP716A141, catalyzed the hydroxylation of β-amyrin at C-16β. This reaction is unique among triterpene oxidases isolated to date. These results enhance our knowledge of functional variation among CYP716A subfamily enzymes involved in triterpenoid biosynthesis, and provide novel molecular tools for use in synthetic biology to produce triterpenoid saponins with pre-defined structures. [ABSTRACT FROM AUTHOR]

• There are 35 trihelix gene family members identified in Platycodon grandiflorus. • Bioinformatics analysis of members of the trihelix gene family of Platycodon grandiflorus. • Bioinformatics analysis of the PgGT1 gene and its response under abiotic stresses. The trihelix gene family plays an important role in plant growth and abiotic stress responses. Through the analysis of genomic and transcriptome data, 35 trihelix family members were identified for the first time in Platycodon grandiflorus ; they were classified into five subfamilies: GT-1, GT-2, SH4, GTγ, and SIP1. The gene structure, conserved motifs and evolutionary relationships were analyzed. Prediction of physicochemical properties of the 35 trihelix proteins founded, the number of amino acid molecules is between 93 and 960, theoretical isoelectric point is between 4.24 and 9.94, molecular weight is between 9829.77 and 107435.38, 4 proteins among them were stable, and all GRAVY is negative. The full-length cDNA sequence of the PgGT1 gene of the GT-1 subfamily was cloned by PCR. It is a 1165 bp ORF encoding a 387 amino acid protein, with a molecular weight of 43.54 kDa. The predicted subcellular localization of the protein in the nucleus was experimentally verified. After being treated with NaCl, PEG6000, MeJA, ABA, IAA, SA, and ethephon, the expression of PgGT1 gene showed an up-regulated trend except for the roots treated with NaCl and ABA. This study laid a bioinformatics foundation for the research of trihelix gene family and the cultivation of excellent germplasm of P. grandiflorus. [ABSTRACT FROM AUTHOR]

Platycodon grandiflorus is the single genus plant in family Campanulaceae, which is distributed in areas of China, Japan, Korea, and Russia. In this study, we assembled and annotated the complete chloroplast genome of P. grandiflorus. The chloroplast genome of P. grandiflorus is 171,624 bp long, containing a shorter small single-copy region (SSC) of 7,962 bp, a normal large single-copy region (LSC) of 79,690 bp and a longer pair of inverted repeat regions (IRs) of 41,986 bp. The whole chloroplast genome of P. grandiflorus has 146 genes, including 102 protein-coding genes (PCGs), 36 transfer RNA genes (tRNAs), and 8 ribosome RNA genes (rRNAs). The overall nucleotide composition is: A of 30.9%, T of 30.9%, C of 19.3%, and G of 18.9%, with a total G + C content of the chloroplast genome 38.2%. The phylogenetic and genetic analysis showed a close relationship between Platycodon grandiflorus and Codonopsis lanceolata. [ABSTRACT FROM AUTHOR]

Increasing hepatic insulin signaling is found to be an important mechanism of Platycodon grandiflorus root to alleviate metabolic syndrome (MetS) symptoms such as insulin resistance, obesity, hyperlipidemia and hepatic steatosis, but the details are not yet clear. Since the main constituents of Platycodon grandiflorus root were hard to be absorbed by gastrointestinal tract, getting opportunity to interact with gut microbiota, we speculate the gut microorganisms may mediate its effect. Our work aimed to confirm the critical role of gut microbes in the intervention of Platycodon grandiflorus root extract (PRE) on MetS, and investigate the mechanism. Biochemical analyses, glucose tolerance test and hepatic lipidomics analysis were used to evaluate the anti-MetS effect of PRE on high fat diet (HFD) fed mice. Perform 16S rDNA analysis, qPCR analysis and in vitro co-incubation experiment to study its effect on gut microbes, followed by fecal microbiota transplantation (FMT) experiment and antibiotics intervention experiment. Also, the effect of Akkermansia muciniphila treatment on HFD mice was investigated. PRE alleviated lipid accumulation and insulin resistance in HFD mice and remodeled the fecal microbiome. It also increased the gene expression of colonic tight junction proteins, alleviated metabolic endotoxemia and inflammation, so that reduced TNF-α induced hepatic JNK-dependent IRS-1 serine phosphorylation and the impairment of PI3K/PIP3/Akt insulin signaling pathway. A. muciniphila was one of the most significantly enriched microbes by PRE treatment, and its administration to HFD mice showed similar effects to PRE, repairing the gut barrier and activating hepatic PI3K/PIP3/Akt pathway. Finally, anti-MetS effect of PRE could be delivered to FMT recipients, and PRE could not further attenuate MetS in gut microbiota depleted mice. We demonstrated for the first time that PRE alleviated MetS in a gut microbiota dependent manner, and found activation of hepatic insulin signaling mediated by gut A. muciniphila was a potential mechanism of it. [Display omitted] [ABSTRACT FROM AUTHOR]