The rapid and accurate authentication of traditional Chinese medicines(TCMs)has always been a key scientific and technical problem in the field of pharmaceutical analysis.Herein,a novel heating online extraction elect...The rapid and accurate authentication of traditional Chinese medicines(TCMs)has always been a key scientific and technical problem in the field of pharmaceutical analysis.Herein,a novel heating online extraction electrospray ionization mass spectrometry(H-oEESI-MS)was developed for the rapid and direct analysis of extremely complex substances without the requirement for any sample pretreatment or pre-separation steps.The overall molecular profile and fragment structure features of various herbal medicines could be completely captured within 10–15 s,with minimal sample(<0.5 mg)and solvent consumption(<20μL for one sample).Furthermore,a rapid differentiation and authentication strategy for TCMs based on H-oEESI-MS was proposed,including metabolic profile characterization,characteristic marker screening and identification,and multivariate statistical analysis model validation.In an analysis of 52 batches of seven types of Aconitum medicinal materials,20 and 21 key compounds were screened out as the characteristic markers of raw and processed Aconitum herbal medicines,respectively,and the possible structures of all the characteristic markers were comprehensively identified based on Compound Discoverer databases.Finally,multivariate statistical analysis showed that all the different types of herbal medicines were well differentiated and identified(R^(2)X>0.87,R^(2)Y>0.91,and Q^(2)>0.72),which further verified the feasibility and reliability of this comprehensive strategy for the rapid authentication of different TCMs based on H-oEESI-MS.In summary,this rapid authentication strategy realized the ultra-high-throughput,low-cost,and standardized detection of various complex TCMs for the first time,thereby demonstrating wide applicability and value for the development of quality standards for TCMs.展开更多
Fallopia multiflora(Thunb.)Harald,a vine belonging to the Polygonaceae family,is used in traditional medicine.The stilbenes contained in it have significant pharmacological activities in anti-oxidation and anti-aging....Fallopia multiflora(Thunb.)Harald,a vine belonging to the Polygonaceae family,is used in traditional medicine.The stilbenes contained in it have significant pharmacological activities in anti-oxidation and anti-aging.This study describes the assembly of the F.multiflora genome and presents its chromosome-level genome sequence containing 1.46 gigabases of data(with a contig N50 of 1.97 megabases),1.44 gigabases of which was assigned to 11 pseudochromosomes.Comparative genomics confirmed that F.multiflora shared a whole-genome duplication event with Tartary buckwheat and then underwent different transposon evolution after separation.Combining genomics,transcriptomics,and metabolomics data to map a network of associated genes and metabolites,we identified two FmRS genes responsible for the catalysis of one molecule of p-coumaroyl-CoA and three molecules of malonyl-CoA to resveratrol in F.multiflora.These findings not only serve as the basis for revealing the stilbene biosynthetic pathway but will also contribute to the development of tools for increasing the production of bioactive stilbenes through molecular breeding in plants or metabolic engineering in microbes.Moreover,the reference genome of F.multiflora is a useful addition to the genomes of the Polygonaceae family.展开更多
Moutan Cortex (MC) has been demonstrated to have an inhibitive effect on inflammation and oxidative stress responses in mesangial cells in our previous study. However, little is known about the components of MC contri...Moutan Cortex (MC) has been demonstrated to have an inhibitive effect on inflammation and oxidative stress responses in mesangial cells in our previous study. However, little is known about the components of MC contributing to this benefit. In the present study, cell membrane immobilized chromatography (CMC), a fast and useful method, was presented for screening potential active components of MC. HBZY-1 cells were incubated with MC (200 μg/mL) at the optimal incubation time (90 min). HPLC-DAD analysis and LC/ESI/MS/MS were performed to distinguish the active components and identify its structural ion fragments. The results showed that eight components binding to HBZY-1 cells were mudanoside B, paeoniflorin sulfonate, paeoniflorin, tetragalloyl glucose (isomeride), hexagalloyl glucose, mudanopiside A, and paeonol. In conclusion, our established CMC might be a useful method for screening potential active components in complicated traditional Chinese medicines. These components might be associated with the efficacy of MC on prevention and treatment of diabetic nephropathy.展开更多
Objective:To comprehensively review the literature relating to the use of Artemisiae Argyi Folium (AAF;the dried leaf of Artemisia argyi Lévl.et Vant.from the Asteraceae family),and qi ai (AAF grown in Qichun Cou...Objective:To comprehensively review the literature relating to the use of Artemisiae Argyi Folium (AAF;the dried leaf of Artemisia argyi Lévl.et Vant.from the Asteraceae family),and qi ai (AAF grown in Qichun County and the adjacent areas in Hubei Province,China) in the field of traditional Chinese medicine.Furthermore,this study aimed to give results of the contents determination of three bioactive components in qi ai and AAF.Methods:The literature was reviewed to acquire relevant information regarding the quality and application of AAF and qi ai.In addition,the contents of the bioactive components (essential oil,total flavonoids and tannins) were determined in 29 AAF samples collected from China and Korea using the methods in the Chinese pharmacopoeia (2015) or determined by our experiments.Results:The dried leaf of A.argyi Lévl.et Vant.from the Asteraceae family is a mainstream commodity of AAF.Qi ai is a commercial type of AAF well-known for its good quality,which has been recorded in Chinese ancient literature.Modern research has showed that more intense heat with stronger penetration was generated in moxibustion therapy when qi ai moxa stick was burnt compared with general moxa stick,which led to a better curative effect.Our research also showed that the contents of essential oil,total flavonoids and tannin in qi ai were significantly higher than those of general AAF samples.Conclusion:The superior quality of qi ai compared with general AAF may be due to relatively higher contents of bioactive components.This study provides basic information for further study and utilization of AAF and qi ai.展开更多
In this paper, electronic nose (E-nose) was used to discriminate the 11 Chinese Materia Medica (CMM) from Umbelliferae by the difference of their odors. The E-nose generated data were analyzed by discriminant function...In this paper, electronic nose (E-nose) was used to discriminate the 11 Chinese Materia Medica (CMM) from Umbelliferae by the difference of their odors. The E-nose generated data were analyzed by discriminant function analysis (DFA) and the responses of 18 sensors of E-nose were evaluated by CA analysis. Results showed that a rapid evaluation of complex response of the samples could be obtained, in combination with DFA, SQC and the CA analysis of the E-nose was given better results in the recognition values of the odor of the 11 CMM. All the 11 CMM could be distinguished by E-nose coupled with DFA, sensor 2, 3, 4, 5, 11, 13 and 15 were found to be able to better discriminate between the CMM samples. The CMM from Umbelliferae produced from different areas and processed with different methods could be distinguished by the E-nose, too. The results of the similarity of fingerprints of the E-nose are fitted with the TCM records about the property (yaoxing), channel tropism (guijing), function and usage of the CMM. The E-nose is a technology that can reflect the holistic odor of a CMM and is relevant to the TCM doctor’s practical identification. The odor of CMM can be expressed by objective data instead of subjective sense by human nose. Based on the sensor`s intensity of E-nose, the fingerprint of a CMM can be established, too. Although the E-nose has so many advantages, only use E-nose technology is not enough to control the quality of a CMM. It must be combined with the other macroscopic discriminating technology, such as the E-tongue, the E-eye, to have a holistic evaluation of a CMM.展开更多
Gut-liver-brain axis is a three-way highway of information interaction system among the gastrointestinal tract,liver,and nervous systems.In the past few decades,breakthrough progress has been made in the gut liver bra...Gut-liver-brain axis is a three-way highway of information interaction system among the gastrointestinal tract,liver,and nervous systems.In the past few decades,breakthrough progress has been made in the gut liver brain axis,mainly through understanding its formation mechanism and increasing treatment strategies.In this review,we discuss various complex networks including barrier permeability,gut hormones,gut microbial metabolites,vagus nerve,neurotransmitters,immunity,brain toxic metabolites,β-amyloid(Aβ)metabolism,and epigenetic regulation in the gut-liver-brain axis.Some therapies containing antibiotics,probiotics,prebiotics,synbiotics,fecal microbiota transplantation(FMT),polyphenols,low FODMAP diet and nanotechnology application regulate the gut liver brain axis.Besides,some special treatments targeting gut-liver axis include farnesoid X receptor(FXR)agonists,takeda G protein-coupled receptor 5(TGR5)agonists,glucagon-like peptide-1(GLP-1)receptor antagonists and fibroblast growth factor 19(FGF19)analogs.Targeting gut-brain axis embraces cognitive behavioral therapy(CBT),antidepressants and tryptophan metabolism-related therapies.Targeting liver-brain axis contains epigenetic regulation and Aβmetabolism-related therapies.In the future,a better understanding of gut-liver-brain axis interactions will promote the development of novel preventative strategies and the discovery of precise therapeutic targets in multiple diseases.展开更多
Aporphine alkaloids have diverse pharmacological activities;however,our understanding of their biosynthesis is relatively limited.Previous studies have classified aporphine alkaloids into two categories based on the c...Aporphine alkaloids have diverse pharmacological activities;however,our understanding of their biosynthesis is relatively limited.Previous studies have classified aporphine alkaloids into two categories based on the configuration and number of substituents of the D-ring and have proposed preliminary biosynthetic pathways for each category.In this study,we identified two specific cytochrome P450 enzymes(CYP80G6 and CYP80Q5)with distinct activities toward(S)-configured and(R)-configured substrates from the herbaceous perennial vine Stephania tetrandra,shedding light on the biosynthetic mechanisms and stereochemical features of these two aporphine alkaloid categories.Additionally,we characterized two CYP719C enzymes(CYP719C3 and CYP719C4)that catalyzed the formation of the methylenedioxy bridge,an essential pharmacophoric group,on the A-and D-rings,respectively,of aporphine alkaloids.Leveraging the functional characterization of these crucial cytochrome P450 enzymes,we reconstructed the biosynthetic pathways for the two types of aporphine alkaloids in budding yeast(Saccharomyces cerevisiae)for the de novo production of compounds such as(R)-glaziovine,(S)-glaziovine,and magnoflorine.This study provides key insight into the biosynthesis of aporphine alkaloids and lays a foundation for producing these valuable compounds through synthetic biology.展开更多
Cytochrome P450 enzymes play a crucial role as catalysts in the biosynthesis of numerous plant natural products(PNPs).Enhancing the catalytic activity of P450s in host microorganisms is essential for the efficient pro...Cytochrome P450 enzymes play a crucial role as catalysts in the biosynthesis of numerous plant natural products(PNPs).Enhancing the catalytic activity of P450s in host microorganisms is essential for the efficient production of PNPs through synthetic biology.In this study,we engineered Saccharomyces cerevisiae to optimize the microenvironment for boosting the activities of P450s,including coexpression with the redox partner genes,enhancing NADPH supply,expanding the endoplasmic reticulum(ER),strengthening heme biosynthesis,and regulating iron uptake.This created a platform for the efficient production 11,20-dihydroxyferruginol,a key intermediate of the bioactive compound tanshinones.The yield was enhanced by 42.1-fold through 24 effective genetic edits.The optimized strain produced up to 67.69±1.33 mg/L 11,20-dihydroxyferruginol in shake flasks.Our work represents a promising advancement toward constructing yeast cell factories containing P450s and paves the way for microbial biosynthesis of tanshinones in the future.展开更多
Vascular complications of diabetes pose a severe threat to human health.Prevention and treatment protocols based on a single vascular complication are no longer suitable for the long-term management of patients with d...Vascular complications of diabetes pose a severe threat to human health.Prevention and treatment protocols based on a single vascular complication are no longer suitable for the long-term management of patients with diabetes.Diabetic panvascular disease(DPD)is a clinical syndrome in which vessels of various sizes,including macrovessels and microvessels in the cardiac,cerebral,renal,ophthalmic,and peripheral systems of patients with diabetes,develop atherosclerosis as a common pathology.Pathological manifestations of DPDs usually manifest macrovascular atherosclerosis,as well as microvascular endothelial function impairment,basement membrane thickening,and microthrombosis.Cardiac,cerebral,and peripheral microangiopathy coexist with microangiopathy,while renal and retinal are predominantly microangiopathic.The following associations exist between DPDs:numerous similar molecular mechanisms,and risk-predictive relationships between diseases.Aggressive glycemic control combined with early comprehensive vascular intervention is the key to prevention and treatment.In addition to the widely recommended metformin,glucagon-like peptide-1 agonist,and sodium-glucose cotransporter-2 inhibitors,for the latest molecular mechanisms,aldose reductase inhibitors,peroxisome proliferator-activated receptor-γagonizts,glucokinases agonizts,mitochondrial energy modulators,etc.are under active development.DPDs are proposed for patients to obtain more systematic clinical care requires a comprehensive diabetes care center focusing on panvascular diseases.This would leverage the advantages of a cross-disciplinary approach to achieve better integration of the pathogenesis and therapeutic evidence.Such a strategy would confer more clinical benefits to patients and promote the comprehensive development of DPD as a discipline.展开更多
For wild natural medicine,unanticipated biodiversity as species or varieties with similar morphological characteristics and sympatric distribution may co-exist in a single batch of medical materials,which affects the ...For wild natural medicine,unanticipated biodiversity as species or varieties with similar morphological characteristics and sympatric distribution may co-exist in a single batch of medical materials,which affects the efficacy and safety of clinical medication.DNA barcoding as an effective species identification tool is limited by its low sample throughput nature.In this study,combining DNA minibarcode,DNA metabarcoding and species delimitation method,a novel biological sources consistency evaluation strategy was proposed,and high level of interspecific and intraspecific variations were observed and validated among 5376 Amynthas samples from 19 sampling points regarded as"Guang Dilong"and 25 batches of proprietary Chinese medicines.Besides Amynthas aspergillum as the authentic source,8 other Molecular Operational Taxonomic Units(MOTUs)were elucidated.Significantly,even the subgroups within A.aspergillum revealed here differ significantly on chemical compositions and biological activity.Fortunately,this biodiversity could be controlled when the collection was limited to designated areas,as proved by 2796"decoction pieces"samples.This batch biological identification method should be introduced as a novel concept regarding natural medicine quality control,and to offer guidelines for in-situ conservation and breeding bases construction of wild natural medicine.展开更多
Diterpenoid alkaloids(DAs) have been often utilized in clinical practice due to their analgesic and anti-infammatory properties. Natural DAs are prevalent in the family Ranunculaceae, notably in the Aconitum genus. Ne...Diterpenoid alkaloids(DAs) have been often utilized in clinical practice due to their analgesic and anti-infammatory properties. Natural DAs are prevalent in the family Ranunculaceae, notably in the Aconitum genus. Nevertheless, the evolutionary origin of the biosynthesis pathway responsible for DA production remains unknown.In this study, we successfully assembled a highquality, pseudochromosome-level genome of the DA-rich species Aconitum vilmorinianum(A.vilmorinianum)(5.76 Gb). An A. vilmorinianumspecific whole-genome duplication event was discovered using comparative genomic analysis,which may aid in the evolution of the DA biosynthesis pathway. We identified several genes involved in DA biosynthesis via integrated genomic, transcriptomic, and metabolomic analyses. These genes included enzymes encoding target ent-kaurene oxidases and aminotransferases, which facilitated the activation of diterpenes and insertion of nitrogen atoms into diterpene skeletons, thereby mediating the transformation of diterpenes into DAs. The divergence periods of these genes in A. vilmorinianum were further assessed, and it was shown that two major types of genes were involved in the establishment of the DA biosynthesis pathway. Our integrated analysis offers fresh insights into the evolutionary origin of DAs in A.vilmorinianum as well as suggestions for engineering the biosynthetic pathways to obtain desired DAs.展开更多
Traditional Chinese medicine(TCM) has played a pivotal role in maintaining the health of Chinese people and is now gaining increasing acceptance around the global scope. However, TCM is confronting more and more conce...Traditional Chinese medicine(TCM) has played a pivotal role in maintaining the health of Chinese people and is now gaining increasing acceptance around the global scope. However, TCM is confronting more and more concerns with respect to its quality. The intrinsic "multicomponent and multitarget" feature of TCM necessitates the establishment of a unique quality and bioactivity evaluation system, which is different from that of the Western medicine. However, TCM is investigated essentially as"herbal medicine" or "natural product", and the pharmacopoeia quality monographs are actually chemicalmarkers-based, which can ensure the consistency only in the assigned chemical markers, but, to some extent, have deviated from the basic TCM theory. A concept of "quality marker"(Q-marker), following the "property-effect-component" theory, is proposed. The establishment of Q-marker integrates multidisciplinary technologies like natural products chemistry, analytical chemistry, bionics, chemometrics,pharmacology, systems biology, and pharmacodynamics, etc. Q-marker-based fingerprint and multicomponent determination conduce to the construction of more scientific quality control system of TCM.This review delineates the background, definition, and properties of Q-marker, and the associated technologies applied for its establishment. Strategies and approaches for establishing Q-marker-based TCM quality control system are presented and highlighted with a few TCM examples.展开更多
Coix is a grass crop domesticated as early as the Neolithic era.It is still widely cultivated for both highly nutritional food and medicinal use.However,the genetic study and breeding of this crop are hindered by the ...Coix is a grass crop domesticated as early as the Neolithic era.It is still widely cultivated for both highly nutritional food and medicinal use.However,the genetic study and breeding of this crop are hindered by the lack of a sequenced genome.Here,we report de novo sequencing and assembly of the 1619-Mb genome of Coix,and annotation of 75.39%repeats and 39629 protein-coding genes.Comparative genomics analysis showed that Coix is more closely related to sorghum than maize,but intriguingly only Coix and maize had a recent genome duplication event,which was not detected in sorghum.We further constructed a genetic map and mapped several important traits,especially the strength of hull.Selection of papery hull(thin:easy dehulling)from the stony hull(thick:difficult dehulling)in wild progenitors was a key step in Coix domestication.The papery hull makes seed easier to process and germinate.Anatomic and global transcriptome analysis revealed that the papery hull is a result of inhibition of cell division and wall biogenesis.We also successfully demonstrated that seed hull pressure resistance is controlled by two major quantitative trait loci(QTLs),which are associated with hull thickness and color,respectively.The two QTLs were further fine mapped within intervals of 250 kb and 146 kb,respectively.These resources provide a platform for evolutionary studies and will facilitate molecular breeding of this important crop.展开更多
Effective visualization of features generated in next-generation sequencing or other experimental assays is of great interest.These features include operational taxonomy units(OTUs)in microbiome analysis(Zhang et al.,...Effective visualization of features generated in next-generation sequencing or other experimental assays is of great interest.These features include operational taxonomy units(OTUs)in microbiome analysis(Zhang et al.,2019),genes,single nucleotide variants,and gene ontology information in transcriptome,genome,and epigenome analysis(Liu et al.,2018),ingredients in traditional Chinese formulas(Li et al.,2020).展开更多
Clinical manifestations of symptoms play a crucial role in the diagnosis and appropriate treatment of diseases and are considered one of the main clinical features for contemporary disease taxonomy(i.e.,international ...Clinical manifestations of symptoms play a crucial role in the diagnosis and appropriate treatment of diseases and are considered one of the main clinical features for contemporary disease taxonomy(i.e.,international classification of diseases,ICD)[1].Deep investigation on molecular connections among symptoms is one of the key tasks for developing a disease-specific knowledge network and thus promoting the refinement of disease taxonomy toward precision medicine[2].展开更多
The effects of different cadmium （Cd） concentrations （0, 20, 60, and 100 μmol/L） on hydroponically grown Artemisia annua L. were investigated. Cd treatments applied for 0, 4, 12, 24, 72, 144, 216, and 336 hr were...The effects of different cadmium （Cd） concentrations （0, 20, 60, and 100 μmol/L） on hydroponically grown Artemisia annua L. were investigated. Cd treatments applied for 0, 4, 12, 24, 72, 144, 216, and 336 hr were assessed by measuring the changes in photosynthetic pigments, electrolyte leakage, malondialdehyde （MDA） and antioxidants （ascorbic acid and glutathione）, while the artemisinin content was tested after 0, 12, 144, 216, and 336 hr. A significant decrease was observed in photosynthetic pigment levels over time with increasing Cd concentration. Chlorophyll b levels were more affected by Cd than were chlorophyll a or carotenoid levels. The cell membrane was sensitive to Cd stress, as MDA content in all treatment groups showed insignificant differences from the control group, except at 12 hr treatment time. Ascorbic acid （AsA） content changed slightly over time, while glutathione （GSH） content took less time to reach a maximum as Cd concentration increased. Cd was found to promote synthesis and accumulation of artemisinin, especially at concentrations of 20 and 100 ~tmol/L. In conclusion, Cd stress can damage to photosynthetic pigments, and vigorously growing A. annua showed a strong tolerance for Cd stress. Appropriate amounts of added Cd aided synthesis and accumulation of artemisinin.展开更多
Aconitum carmichaelii is a high-value medicinal herb widely used across China,Japan,and other Asian countries.Aconitine-type diterpene alkaloids(DAs)are the characteristic compounds in Aconitum.Although six transcript...Aconitum carmichaelii is a high-value medicinal herb widely used across China,Japan,and other Asian countries.Aconitine-type diterpene alkaloids(DAs)are the characteristic compounds in Aconitum.Although six transcriptomes,based on short-read next generation sequencing technology,have been reported from the Aconitum species,the terpene synthase(TPS)corresponding to DAs biosynthesis remains unidentified.We apply a combination of Pacbio isoform sequencing and RNA sequencing to provide a comprehensive view of the A.carmichaelii transcriptome.Nineteen TPSs and five alternative splicing isoforms belonging to TPS-b,TPS-c,and TPS-e/f subfamilies were identified.In vitro enzyme reaction analysis functional identified two sesqui-TPSs and twelve di TPSs.Seven of the TPS-c subfamily genes reacted with GGPP to produce the intermediate ent-copalyl diphosphate.Five Ac KSLs separately reacted with ent-CPP to produce ent-kaurene,ent-atiserene,and ent-13-epi-sandaracopimaradie:a new diterpene found in Aconitum.Ac TPSs gene expression in conjunction DAs content analysis in different tissues validated that ent-CPP is the sole precursor to all DAs biosynthesis,with Ac KSL1,Ac KSL2 s and Ac KSL3-1 responsible for C20 atisine and napelline type DAs biosynthesis,respectively.These data clarified the molecular basis for the C20-DAs biosynthetic pathway in A.carmichaelii and pave the way for further exploration of C19-DAs biosynthesis in the Aconitum species.展开更多
Panax notoginseng,a perennial herb of the genus Panax in the family Araliaceae,has played an important role in clinical treatment in China for thousands of years because of its extensive pharmacological effects.Here,w...Panax notoginseng,a perennial herb of the genus Panax in the family Araliaceae,has played an important role in clinical treatment in China for thousands of years because of its extensive pharmacological effects.Here,we report a high-quality reference genome of P.notoginseng,with a genome size up to 2.66 Gb and a contig N50 of 1.12 Mb,produced with third-generation PacBio sequencing technology.This is the first chromosome-level genome assembly for the genus Panax.Through genome evolution analysis,we explored phylogenetic and whole-genome duplication events and examined their impact on saponin biosynthesis.We performed a detailed transcriptional analysis of P.notoginseng and explored genelevel mechanisms that regulate the formation of characteristic tubercles.Next,we studied the biosynthesis and regulation of saponins at temporal and spatial levels.We combined multi-omics data to identify genes that encode key enzymes in the P.notoginseng terpenoid biosynthetic pathway.Finally,we identified five glycosyltransferase genes whose products catalyzed the formation of different ginsenosides in P.notoginseng.The genetic information obtained in this study provides a resource for further exploration of the growth characteristics,cultivation,breeding,and saponin biosynthesis of P.notoginseng.展开更多
Sterol C24-methyltransferase(SMT) plays multiple important roles in plant growth and development. SMT1, which belongs to the family of transferases and transforms cycloartenol into 24-methylene cycloartenol, is involv...Sterol C24-methyltransferase(SMT) plays multiple important roles in plant growth and development. SMT1, which belongs to the family of transferases and transforms cycloartenol into 24-methylene cycloartenol, is involved in the biosynthesis of 24-methyl sterols. Here, we report the cloning and characterization of a cDNA encoding a sterol C24-methyltransferase from Tripterygium wilfordii(Tw SMT1). Tw SMT1(Gen Bank access number KU885950) is a 1530 bp cDNA with a 1041 bp open reading frame predicted to encode a 346-amino acid, 38.62 k Da protein. The polypeptide encoded by the SMT1 cDNA was expressed and purified as a recombinant protein from Escherichia coli(E. coli) and showed SMT activity. The expression of Tw SMT1 was highly up-regulated in T. wilfordii cell suspension cultures treated with methyl jasmonate(Me JA). Tissue expression pattern analysis showed higher expression in the phellem layer compared to the other four organs(leaf, stem, xylem and phloem), which is about ten times that of the lowest expression in leaf. The results are meaningful for the study of sterolbiosynthesis of T. wilfordii and will further lay the foundations for the research in regulating both the content of other main compounds and growth and development of T. wilfordii.展开更多
The ginsenoside Rgfound in Panax species has extensive pharmacological properties,in particular anti-cancer effects.However,its natural yield in Panax plants is limited.Here,we report a multimodular strategy to improv...The ginsenoside Rgfound in Panax species has extensive pharmacological properties,in particular anti-cancer effects.However,its natural yield in Panax plants is limited.Here,we report a multimodular strategy to improve yields of Rgin a Panax ginseng chassis,combining engineering of triterpene metabolism and overexpression of a lignin biosynthesis gene,phenylalanine ammonia lyase(PAL).We first performed semi-rational design and site mutagenesis to improve the enzymatic efficiency of Pq3-O-UGT2,a glycosyltransferase that directly catalyzes the biosynthesis of Rgfrom Rh.Next,we used clustered regularly interspaced palindromic repeats(CRISPR)/CRISPR-associated protein 9(Cas9)gene editing to knock down the branch pathway of protopanaxatriol-type ginsenoside biosynthesis to enhance the metabolic flux of the protopanaxadiol-type ginsenoside Rg.Overexpression of PAL accelerated the formation of the xylem structure,significantly improving ginsenoside Rgaccumulation(to 6.19-fold higher than in thecontrol).Wecombinedoverexpression of the ginsenoside aglycon synthetic genes squalene epoxidase,Pq3-O-UGT2,and PAL with CRISPR/Cas9-based knockdown of CYP716A53v2 to improve ginsenoside Rgaccumulation.Finally,we produced ginsenoside Rgat a yield of 83.6 mg/L in a shake flask(7.0 mg/g dry weight,21.12-fold higher than with wild-type cultures).The highproduction system established in this study could be a potential platform to produce the ginsenoside Rgcommercially for pharmaceutical use.展开更多
基金supported by the CACMS Innovation Fund,China(Grant Nos.:CI2021A04504 and CI2021A05206)the National Natural Science Foundation of China(Grant Nos.:82104380,81891010,81891013,and 82074012)+2 种基金the Fundamental Research Funds for the Central Public Welfare Research Institutes,China(Grant Nos.:ZZ14-YQ-047 and ZZXT202105)the Key Project at Central Government Level(Grant No.:2060302-2201-26)the Beijing Nova Program.
文摘The rapid and accurate authentication of traditional Chinese medicines(TCMs)has always been a key scientific and technical problem in the field of pharmaceutical analysis.Herein,a novel heating online extraction electrospray ionization mass spectrometry(H-oEESI-MS)was developed for the rapid and direct analysis of extremely complex substances without the requirement for any sample pretreatment or pre-separation steps.The overall molecular profile and fragment structure features of various herbal medicines could be completely captured within 10–15 s,with minimal sample(<0.5 mg)and solvent consumption(<20μL for one sample).Furthermore,a rapid differentiation and authentication strategy for TCMs based on H-oEESI-MS was proposed,including metabolic profile characterization,characteristic marker screening and identification,and multivariate statistical analysis model validation.In an analysis of 52 batches of seven types of Aconitum medicinal materials,20 and 21 key compounds were screened out as the characteristic markers of raw and processed Aconitum herbal medicines,respectively,and the possible structures of all the characteristic markers were comprehensively identified based on Compound Discoverer databases.Finally,multivariate statistical analysis showed that all the different types of herbal medicines were well differentiated and identified(R^(2)X>0.87,R^(2)Y>0.91,and Q^(2)>0.72),which further verified the feasibility and reliability of this comprehensive strategy for the rapid authentication of different TCMs based on H-oEESI-MS.In summary,this rapid authentication strategy realized the ultra-high-throughput,low-cost,and standardized detection of various complex TCMs for the first time,thereby demonstrating wide applicability and value for the development of quality standards for TCMs.
基金This study was funded by the National Natural Science Foundation of China(No.81973432)the Major Increase and Reduction Project at the Central Level(No.2060302)+1 种基金Innovation Team and Talents Cultivation Program of National Administration of Traditional Chinese Medicine(ZYYCXTD-D-202005)CAMS Innovation Fund for Medical Sciences(2019-I2M-5-065).
文摘Fallopia multiflora(Thunb.)Harald,a vine belonging to the Polygonaceae family,is used in traditional medicine.The stilbenes contained in it have significant pharmacological activities in anti-oxidation and anti-aging.This study describes the assembly of the F.multiflora genome and presents its chromosome-level genome sequence containing 1.46 gigabases of data(with a contig N50 of 1.97 megabases),1.44 gigabases of which was assigned to 11 pseudochromosomes.Comparative genomics confirmed that F.multiflora shared a whole-genome duplication event with Tartary buckwheat and then underwent different transposon evolution after separation.Combining genomics,transcriptomics,and metabolomics data to map a network of associated genes and metabolites,we identified two FmRS genes responsible for the catalysis of one molecule of p-coumaroyl-CoA and three molecules of malonyl-CoA to resveratrol in F.multiflora.These findings not only serve as the basis for revealing the stilbene biosynthetic pathway but will also contribute to the development of tools for increasing the production of bioactive stilbenes through molecular breeding in plants or metabolic engineering in microbes.Moreover,the reference genome of F.multiflora is a useful addition to the genomes of the Polygonaceae family.
文摘Moutan Cortex (MC) has been demonstrated to have an inhibitive effect on inflammation and oxidative stress responses in mesangial cells in our previous study. However, little is known about the components of MC contributing to this benefit. In the present study, cell membrane immobilized chromatography (CMC), a fast and useful method, was presented for screening potential active components of MC. HBZY-1 cells were incubated with MC (200 μg/mL) at the optimal incubation time (90 min). HPLC-DAD analysis and LC/ESI/MS/MS were performed to distinguish the active components and identify its structural ion fragments. The results showed that eight components binding to HBZY-1 cells were mudanoside B, paeoniflorin sulfonate, paeoniflorin, tetragalloyl glucose (isomeride), hexagalloyl glucose, mudanopiside A, and paeonol. In conclusion, our established CMC might be a useful method for screening potential active components in complicated traditional Chinese medicines. These components might be associated with the efficacy of MC on prevention and treatment of diabetic nephropathy.
文摘Objective:To comprehensively review the literature relating to the use of Artemisiae Argyi Folium (AAF;the dried leaf of Artemisia argyi Lévl.et Vant.from the Asteraceae family),and qi ai (AAF grown in Qichun County and the adjacent areas in Hubei Province,China) in the field of traditional Chinese medicine.Furthermore,this study aimed to give results of the contents determination of three bioactive components in qi ai and AAF.Methods:The literature was reviewed to acquire relevant information regarding the quality and application of AAF and qi ai.In addition,the contents of the bioactive components (essential oil,total flavonoids and tannins) were determined in 29 AAF samples collected from China and Korea using the methods in the Chinese pharmacopoeia (2015) or determined by our experiments.Results:The dried leaf of A.argyi Lévl.et Vant.from the Asteraceae family is a mainstream commodity of AAF.Qi ai is a commercial type of AAF well-known for its good quality,which has been recorded in Chinese ancient literature.Modern research has showed that more intense heat with stronger penetration was generated in moxibustion therapy when qi ai moxa stick was burnt compared with general moxa stick,which led to a better curative effect.Our research also showed that the contents of essential oil,total flavonoids and tannin in qi ai were significantly higher than those of general AAF samples.Conclusion:The superior quality of qi ai compared with general AAF may be due to relatively higher contents of bioactive components.This study provides basic information for further study and utilization of AAF and qi ai.
文摘In this paper, electronic nose (E-nose) was used to discriminate the 11 Chinese Materia Medica (CMM) from Umbelliferae by the difference of their odors. The E-nose generated data were analyzed by discriminant function analysis (DFA) and the responses of 18 sensors of E-nose were evaluated by CA analysis. Results showed that a rapid evaluation of complex response of the samples could be obtained, in combination with DFA, SQC and the CA analysis of the E-nose was given better results in the recognition values of the odor of the 11 CMM. All the 11 CMM could be distinguished by E-nose coupled with DFA, sensor 2, 3, 4, 5, 11, 13 and 15 were found to be able to better discriminate between the CMM samples. The CMM from Umbelliferae produced from different areas and processed with different methods could be distinguished by the E-nose, too. The results of the similarity of fingerprints of the E-nose are fitted with the TCM records about the property (yaoxing), channel tropism (guijing), function and usage of the CMM. The E-nose is a technology that can reflect the holistic odor of a CMM and is relevant to the TCM doctor’s practical identification. The odor of CMM can be expressed by objective data instead of subjective sense by human nose. Based on the sensor`s intensity of E-nose, the fingerprint of a CMM can be established, too. Although the E-nose has so many advantages, only use E-nose technology is not enough to control the quality of a CMM. It must be combined with the other macroscopic discriminating technology, such as the E-tongue, the E-eye, to have a holistic evaluation of a CMM.
基金The National Natural Science Foundation of China(82074069)Tianjin Municipal Science and Technology Committee(20YFZCSY00560)funded this work.
文摘Gut-liver-brain axis is a three-way highway of information interaction system among the gastrointestinal tract,liver,and nervous systems.In the past few decades,breakthrough progress has been made in the gut liver brain axis,mainly through understanding its formation mechanism and increasing treatment strategies.In this review,we discuss various complex networks including barrier permeability,gut hormones,gut microbial metabolites,vagus nerve,neurotransmitters,immunity,brain toxic metabolites,β-amyloid(Aβ)metabolism,and epigenetic regulation in the gut-liver-brain axis.Some therapies containing antibiotics,probiotics,prebiotics,synbiotics,fecal microbiota transplantation(FMT),polyphenols,low FODMAP diet and nanotechnology application regulate the gut liver brain axis.Besides,some special treatments targeting gut-liver axis include farnesoid X receptor(FXR)agonists,takeda G protein-coupled receptor 5(TGR5)agonists,glucagon-like peptide-1(GLP-1)receptor antagonists and fibroblast growth factor 19(FGF19)analogs.Targeting gut-brain axis embraces cognitive behavioral therapy(CBT),antidepressants and tryptophan metabolism-related therapies.Targeting liver-brain axis contains epigenetic regulation and Aβmetabolism-related therapies.In the future,a better understanding of gut-liver-brain axis interactions will promote the development of novel preventative strategies and the discovery of precise therapeutic targets in multiple diseases.
基金supported by the National Key R&D Program of China(2020YFA0908000)the National Natural Science Foundation of China(82011530137,31961133007)+2 种基金Scientific and technological innovation project of CACMS(CI2023D002,CI2023E002)Key project at central government level:The ability to establish sustainable use of valuable Chinese medicine resources(2060302)Vetenskapsradet(2018-06003),Stiftelsen for internationalisering av hogre utbildning och forskning。
文摘Aporphine alkaloids have diverse pharmacological activities;however,our understanding of their biosynthesis is relatively limited.Previous studies have classified aporphine alkaloids into two categories based on the configuration and number of substituents of the D-ring and have proposed preliminary biosynthetic pathways for each category.In this study,we identified two specific cytochrome P450 enzymes(CYP80G6 and CYP80Q5)with distinct activities toward(S)-configured and(R)-configured substrates from the herbaceous perennial vine Stephania tetrandra,shedding light on the biosynthetic mechanisms and stereochemical features of these two aporphine alkaloid categories.Additionally,we characterized two CYP719C enzymes(CYP719C3 and CYP719C4)that catalyzed the formation of the methylenedioxy bridge,an essential pharmacophoric group,on the A-and D-rings,respectively,of aporphine alkaloids.Leveraging the functional characterization of these crucial cytochrome P450 enzymes,we reconstructed the biosynthetic pathways for the two types of aporphine alkaloids in budding yeast(Saccharomyces cerevisiae)for the de novo production of compounds such as(R)-glaziovine,(S)-glaziovine,and magnoflorine.This study provides key insight into the biosynthesis of aporphine alkaloids and lays a foundation for producing these valuable compounds through synthetic biology.
基金supported by the National Key R&D Program of China(2020YFA0908000)Scientific and technological innovation project of China Academy of Chinese Medical Sciences(CI2023D002,CI2023E002,CI2021A04110,CI2021B014)+1 种基金the National Natural Science Foundation of China(81822046)Key project at central government level:The ability to establish sustainable use of valuable Chinese medicine resources(2060302).
文摘Cytochrome P450 enzymes play a crucial role as catalysts in the biosynthesis of numerous plant natural products(PNPs).Enhancing the catalytic activity of P450s in host microorganisms is essential for the efficient production of PNPs through synthetic biology.In this study,we engineered Saccharomyces cerevisiae to optimize the microenvironment for boosting the activities of P450s,including coexpression with the redox partner genes,enhancing NADPH supply,expanding the endoplasmic reticulum(ER),strengthening heme biosynthesis,and regulating iron uptake.This created a platform for the efficient production 11,20-dihydroxyferruginol,a key intermediate of the bioactive compound tanshinones.The yield was enhanced by 42.1-fold through 24 effective genetic edits.The optimized strain produced up to 67.69±1.33 mg/L 11,20-dihydroxyferruginol in shake flasks.Our work represents a promising advancement toward constructing yeast cell factories containing P450s and paves the way for microbial biosynthesis of tanshinones in the future.
基金supported by the National Outstanding Youth Natural Science Foundation of China (82022076)the Young Qihuang Scholar of the“Tens of millions”talent project of China.
文摘Vascular complications of diabetes pose a severe threat to human health.Prevention and treatment protocols based on a single vascular complication are no longer suitable for the long-term management of patients with diabetes.Diabetic panvascular disease(DPD)is a clinical syndrome in which vessels of various sizes,including macrovessels and microvessels in the cardiac,cerebral,renal,ophthalmic,and peripheral systems of patients with diabetes,develop atherosclerosis as a common pathology.Pathological manifestations of DPDs usually manifest macrovascular atherosclerosis,as well as microvascular endothelial function impairment,basement membrane thickening,and microthrombosis.Cardiac,cerebral,and peripheral microangiopathy coexist with microangiopathy,while renal and retinal are predominantly microangiopathic.The following associations exist between DPDs:numerous similar molecular mechanisms,and risk-predictive relationships between diseases.Aggressive glycemic control combined with early comprehensive vascular intervention is the key to prevention and treatment.In addition to the widely recommended metformin,glucagon-like peptide-1 agonist,and sodium-glucose cotransporter-2 inhibitors,for the latest molecular mechanisms,aldose reductase inhibitors,peroxisome proliferator-activated receptor-γagonizts,glucokinases agonizts,mitochondrial energy modulators,etc.are under active development.DPDs are proposed for patients to obtain more systematic clinical care requires a comprehensive diabetes care center focusing on panvascular diseases.This would leverage the advantages of a cross-disciplinary approach to achieve better integration of the pathogenesis and therapeutic evidence.Such a strategy would confer more clinical benefits to patients and promote the comprehensive development of DPD as a discipline.
基金supported by Foundation Science and Technology Program of Tianjin(No.22ZYJDSS00040,20ZYJDJC00120,China)Foundation CACMS Innovation Fund(No.CI2021A04104,China)Foundation Key project at central government level:The ability establishment of sustainable use for valuable Chinese medicine resources(No.2060302,China)。
文摘For wild natural medicine,unanticipated biodiversity as species or varieties with similar morphological characteristics and sympatric distribution may co-exist in a single batch of medical materials,which affects the efficacy and safety of clinical medication.DNA barcoding as an effective species identification tool is limited by its low sample throughput nature.In this study,combining DNA minibarcode,DNA metabarcoding and species delimitation method,a novel biological sources consistency evaluation strategy was proposed,and high level of interspecific and intraspecific variations were observed and validated among 5376 Amynthas samples from 19 sampling points regarded as"Guang Dilong"and 25 batches of proprietary Chinese medicines.Besides Amynthas aspergillum as the authentic source,8 other Molecular Operational Taxonomic Units(MOTUs)were elucidated.Significantly,even the subgroups within A.aspergillum revealed here differ significantly on chemical compositions and biological activity.Fortunately,this biodiversity could be controlled when the collection was limited to designated areas,as proved by 2796"decoction pieces"samples.This batch biological identification method should be introduced as a novel concept regarding natural medicine quality control,and to offer guidelines for in-situ conservation and breeding bases construction of wild natural medicine.
基金supported by funding from the CAS “Pioneer Hundred Talents” Program and Strategic Priority Research Program of the Chinese Academy of Sciences (XDB31000000) to Y.C.National Natural Science Foundation of China (Nos. 31960082, 32270395, and 31960096)Yunnan Fundamental Research Projects (No. 202101AS070021) to D.Z。
文摘Diterpenoid alkaloids(DAs) have been often utilized in clinical practice due to their analgesic and anti-infammatory properties. Natural DAs are prevalent in the family Ranunculaceae, notably in the Aconitum genus. Nevertheless, the evolutionary origin of the biosynthesis pathway responsible for DA production remains unknown.In this study, we successfully assembled a highquality, pseudochromosome-level genome of the DA-rich species Aconitum vilmorinianum(A.vilmorinianum)(5.76 Gb). An A. vilmorinianumspecific whole-genome duplication event was discovered using comparative genomic analysis,which may aid in the evolution of the DA biosynthesis pathway. We identified several genes involved in DA biosynthesis via integrated genomic, transcriptomic, and metabolomic analyses. These genes included enzymes encoding target ent-kaurene oxidases and aminotransferases, which facilitated the activation of diterpenes and insertion of nitrogen atoms into diterpene skeletons, thereby mediating the transformation of diterpenes into DAs. The divergence periods of these genes in A. vilmorinianum were further assessed, and it was shown that two major types of genes were involved in the establishment of the DA biosynthesis pathway. Our integrated analysis offers fresh insights into the evolutionary origin of DAs in A.vilmorinianum as well as suggestions for engineering the biosynthetic pathways to obtain desired DAs.
文摘Traditional Chinese medicine(TCM) has played a pivotal role in maintaining the health of Chinese people and is now gaining increasing acceptance around the global scope. However, TCM is confronting more and more concerns with respect to its quality. The intrinsic "multicomponent and multitarget" feature of TCM necessitates the establishment of a unique quality and bioactivity evaluation system, which is different from that of the Western medicine. However, TCM is investigated essentially as"herbal medicine" or "natural product", and the pharmacopoeia quality monographs are actually chemicalmarkers-based, which can ensure the consistency only in the assigned chemical markers, but, to some extent, have deviated from the basic TCM theory. A concept of "quality marker"(Q-marker), following the "property-effect-component" theory, is proposed. The establishment of Q-marker integrates multidisciplinary technologies like natural products chemistry, analytical chemistry, bionics, chemometrics,pharmacology, systems biology, and pharmacodynamics, etc. Q-marker-based fingerprint and multicomponent determination conduce to the construction of more scientific quality control system of TCM.This review delineates the background, definition, and properties of Q-marker, and the associated technologies applied for its establishment. Strategies and approaches for establishing Q-marker-based TCM quality control system are presented and highlighted with a few TCM examples.
基金supported by the National Natural Science Foundation of China(grant no.31971753,81703657)Key Project at Central Government Level:the Ability Establishment of Sustainable Use for Valuable Chinese Medicine Resources(2060302)+2 种基金the Fundamental Research Funds for Central Non-profit Scientific Institution of China(Y2017XM06,ZZ13-YQ-085)the Key Science and Technology Program of Biotechnology and Medicine of Sichuan Province(2018NZDZX0001)the National Basic Research Program of China(2014CB138705).
文摘Coix is a grass crop domesticated as early as the Neolithic era.It is still widely cultivated for both highly nutritional food and medicinal use.However,the genetic study and breeding of this crop are hindered by the lack of a sequenced genome.Here,we report de novo sequencing and assembly of the 1619-Mb genome of Coix,and annotation of 75.39%repeats and 39629 protein-coding genes.Comparative genomics analysis showed that Coix is more closely related to sorghum than maize,but intriguingly only Coix and maize had a recent genome duplication event,which was not detected in sorghum.We further constructed a genetic map and mapped several important traits,especially the strength of hull.Selection of papery hull(thin:easy dehulling)from the stony hull(thick:difficult dehulling)in wild progenitors was a key step in Coix domestication.The papery hull makes seed easier to process and germinate.Anatomic and global transcriptome analysis revealed that the papery hull is a result of inhibition of cell division and wall biogenesis.We also successfully demonstrated that seed hull pressure resistance is controlled by two major quantitative trait loci(QTLs),which are associated with hull thickness and color,respectively.The two QTLs were further fine mapped within intervals of 250 kb and 146 kb,respectively.These resources provide a platform for evolutionary studies and will facilitate molecular breeding of this important crop.
基金supported by the Fundamental Research Funds for the Central Public Welfare Research Institutes(ZZ13-YQ-095 and ZZXT201708)Youth Innovation Promotion Association CAS(2021092)。
文摘Effective visualization of features generated in next-generation sequencing or other experimental assays is of great interest.These features include operational taxonomy units(OTUs)in microbiome analysis(Zhang et al.,2019),genes,single nucleotide variants,and gene ontology information in transcriptome,genome,and epigenome analysis(Liu et al.,2018),ingredients in traditional Chinese formulas(Li et al.,2020).
基金supported by the National Natural Science Foundation of China(81830111,82030122,82174533,and 81774201)National Key Research and Development Program of China(2018YFC1705201)+2 种基金Innovation Project of China Academy of Chinese Medical Sciences(CI2021A04907)Youth Innovation Team of Shaanxi Universities and Shaanxi Provincial Science and Technology Department Project(2016SF-378)Fundamental Research Funds for the Central Public Welfare Research Institutes(ZXKT17058 and ZZ13-YQ-095)。
文摘Clinical manifestations of symptoms play a crucial role in the diagnosis and appropriate treatment of diseases and are considered one of the main clinical features for contemporary disease taxonomy(i.e.,international classification of diseases,ICD)[1].Deep investigation on molecular connections among symptoms is one of the key tasks for developing a disease-specific knowledge network and thus promoting the refinement of disease taxonomy toward precision medicine[2].
基金supported by the National Natural Science Foundation of China (No. 81130070,81072989)the Research Projects of State Administration of Traditional Chinese Medicine of China (No. 201107009)+1 种基金the Important National Science & Technology Specific Projects (No.2009ZX09502-026,2009ZX09301-005)the Research Projects of China Academy of Chinese Medical Sciences(No. ZZ20090302)
文摘The effects of different cadmium （Cd） concentrations （0, 20, 60, and 100 μmol/L） on hydroponically grown Artemisia annua L. were investigated. Cd treatments applied for 0, 4, 12, 24, 72, 144, 216, and 336 hr were assessed by measuring the changes in photosynthetic pigments, electrolyte leakage, malondialdehyde （MDA） and antioxidants （ascorbic acid and glutathione）, while the artemisinin content was tested after 0, 12, 144, 216, and 336 hr. A significant decrease was observed in photosynthetic pigment levels over time with increasing Cd concentration. Chlorophyll b levels were more affected by Cd than were chlorophyll a or carotenoid levels. The cell membrane was sensitive to Cd stress, as MDA content in all treatment groups showed insignificant differences from the control group, except at 12 hr treatment time. Ascorbic acid （AsA） content changed slightly over time, while glutathione （GSH） content took less time to reach a maximum as Cd concentration increased. Cd was found to promote synthesis and accumulation of artemisinin, especially at concentrations of 20 and 100 ~tmol/L. In conclusion, Cd stress can damage to photosynthetic pigments, and vigorously growing A. annua showed a strong tolerance for Cd stress. Appropriate amounts of added Cd aided synthesis and accumulation of artemisinin.
基金supported by the Major Program of National Natural Science Foundation of China(81891010,81891013)the National Natural Science Foundation of China(81822046)+1 种基金National Key R&D Program of China(2018YFA0900600,2020YFA0908000)Key project at central government level:the ability to establish sustainable use of valuable Chinese Medicine Resources(2060302,China)。
文摘Aconitum carmichaelii is a high-value medicinal herb widely used across China,Japan,and other Asian countries.Aconitine-type diterpene alkaloids(DAs)are the characteristic compounds in Aconitum.Although six transcriptomes,based on short-read next generation sequencing technology,have been reported from the Aconitum species,the terpene synthase(TPS)corresponding to DAs biosynthesis remains unidentified.We apply a combination of Pacbio isoform sequencing and RNA sequencing to provide a comprehensive view of the A.carmichaelii transcriptome.Nineteen TPSs and five alternative splicing isoforms belonging to TPS-b,TPS-c,and TPS-e/f subfamilies were identified.In vitro enzyme reaction analysis functional identified two sesqui-TPSs and twelve di TPSs.Seven of the TPS-c subfamily genes reacted with GGPP to produce the intermediate ent-copalyl diphosphate.Five Ac KSLs separately reacted with ent-CPP to produce ent-kaurene,ent-atiserene,and ent-13-epi-sandaracopimaradie:a new diterpene found in Aconitum.Ac TPSs gene expression in conjunction DAs content analysis in different tissues validated that ent-CPP is the sole precursor to all DAs biosynthesis,with Ac KSL1,Ac KSL2 s and Ac KSL3-1 responsible for C20 atisine and napelline type DAs biosynthesis,respectively.These data clarified the molecular basis for the C20-DAs biosynthetic pathway in A.carmichaelii and pave the way for further exploration of C19-DAs biosynthesis in the Aconitum species.
基金support from the National Natural Science Foundation of China(nos.81891010,81891013)the Key Project at central government level:The ability establishment of sustainable use for valuable Chinese medicine resources(no.2060302-1806-03)+1 种基金the High-level Teachers in Beijing Municipal Universities in the Period of 13th Five-year Plan(no.CIT&TCD20170324)the National Program for Special Support of Eminent Professionals.
文摘Panax notoginseng,a perennial herb of the genus Panax in the family Araliaceae,has played an important role in clinical treatment in China for thousands of years because of its extensive pharmacological effects.Here,we report a high-quality reference genome of P.notoginseng,with a genome size up to 2.66 Gb and a contig N50 of 1.12 Mb,produced with third-generation PacBio sequencing technology.This is the first chromosome-level genome assembly for the genus Panax.Through genome evolution analysis,we explored phylogenetic and whole-genome duplication events and examined their impact on saponin biosynthesis.We performed a detailed transcriptional analysis of P.notoginseng and explored genelevel mechanisms that regulate the formation of characteristic tubercles.Next,we studied the biosynthesis and regulation of saponins at temporal and spatial levels.We combined multi-omics data to identify genes that encode key enzymes in the P.notoginseng terpenoid biosynthetic pathway.Finally,we identified five glycosyltransferase genes whose products catalyzed the formation of different ginsenosides in P.notoginseng.The genetic information obtained in this study provides a resource for further exploration of the growth characteristics,cultivation,breeding,and saponin biosynthesis of P.notoginseng.
基金supported by the National Natural Science Foundation of China (81422053 and 81373906 to Wei Gao and 81325023 to Luqi Huang)the Support Project of High-level Teachers in Beijing Municipal Universities in the Period of 13th Five-year Plan (CIT&TCD20170324 to Wei Gao)the Key project at Central Government Level: The Ability Establishment of Sustainable Use for Valuable Chinese Medicine Resources (2060302 to Luqi Huang)
文摘Sterol C24-methyltransferase(SMT) plays multiple important roles in plant growth and development. SMT1, which belongs to the family of transferases and transforms cycloartenol into 24-methylene cycloartenol, is involved in the biosynthesis of 24-methyl sterols. Here, we report the cloning and characterization of a cDNA encoding a sterol C24-methyltransferase from Tripterygium wilfordii(Tw SMT1). Tw SMT1(Gen Bank access number KU885950) is a 1530 bp cDNA with a 1041 bp open reading frame predicted to encode a 346-amino acid, 38.62 k Da protein. The polypeptide encoded by the SMT1 cDNA was expressed and purified as a recombinant protein from Escherichia coli(E. coli) and showed SMT activity. The expression of Tw SMT1 was highly up-regulated in T. wilfordii cell suspension cultures treated with methyl jasmonate(Me JA). Tissue expression pattern analysis showed higher expression in the phellem layer compared to the other four organs(leaf, stem, xylem and phloem), which is about ten times that of the lowest expression in leaf. The results are meaningful for the study of sterolbiosynthesis of T. wilfordii and will further lay the foundations for the research in regulating both the content of other main compounds and growth and development of T. wilfordii.
基金financial support from the National Key Research and Development Program(2020YFA0907903)a key project at the central government level:“The ability to establish a sustainable use for valuable Chinese medicine resources”(2060302)+2 种基金the National Science Foundation of China(91954112 and 31900501)the Young Elite Scientists Sponsorship Program of Tianjin(TJSQNTJ-2020-19)the Scientific Research Transformation Foundation of the Wenzhou Safety(Emergency)Institute of Tianjin University。
文摘The ginsenoside Rgfound in Panax species has extensive pharmacological properties,in particular anti-cancer effects.However,its natural yield in Panax plants is limited.Here,we report a multimodular strategy to improve yields of Rgin a Panax ginseng chassis,combining engineering of triterpene metabolism and overexpression of a lignin biosynthesis gene,phenylalanine ammonia lyase(PAL).We first performed semi-rational design and site mutagenesis to improve the enzymatic efficiency of Pq3-O-UGT2,a glycosyltransferase that directly catalyzes the biosynthesis of Rgfrom Rh.Next,we used clustered regularly interspaced palindromic repeats(CRISPR)/CRISPR-associated protein 9(Cas9)gene editing to knock down the branch pathway of protopanaxatriol-type ginsenoside biosynthesis to enhance the metabolic flux of the protopanaxadiol-type ginsenoside Rg.Overexpression of PAL accelerated the formation of the xylem structure,significantly improving ginsenoside Rgaccumulation(to 6.19-fold higher than in thecontrol).Wecombinedoverexpression of the ginsenoside aglycon synthetic genes squalene epoxidase,Pq3-O-UGT2,and PAL with CRISPR/Cas9-based knockdown of CYP716A53v2 to improve ginsenoside Rgaccumulation.Finally,we produced ginsenoside Rgat a yield of 83.6 mg/L in a shake flask(7.0 mg/g dry weight,21.12-fold higher than with wild-type cultures).The highproduction system established in this study could be a potential platform to produce the ginsenoside Rgcommercially for pharmaceutical use.
