Naturally occurring yellow leaf mutants are an important resource for studying pigment content and biosynthesis,as well as related gene expression.In our ongoing cultivation of Rehmannia chingii H.L.Li,we found an off...Naturally occurring yellow leaf mutants are an important resource for studying pigment content and biosynthesis,as well as related gene expression.In our ongoing cultivation of Rehmannia chingii H.L.Li,we found an off-type yellow plant.The yellowing started with the new leaves and gradually spread downward until the entire plant exhibited a stable shade of yellow.We studied the differences in the chlorophyll and carotenoid content,carotenoid profile,and transcriptome of this yellow-leaf mutant(P2).Compared to the wild-type R.chingii plant(P1),P2 leaves had significantly lower chlorophyll and carotenoid content.LC-MS/MS analysis revealed that P2 had higher quantities of severalmetabolites in the carotenoid biosynthesis pathway.Transcriptome sequencing results showed that genes involved in porphyrin metabolism,carbon fixation,photosynthesis and antenna proteins,terpenoid backbone biosynthesis,and carotenoid biosynthesis were differentially expressed between P1 and P2.Large-scale expression differences were observed in the phytohormone and MAPK signaling pathways,as well as in 15 transcription factor families.We discuss possible mechanisms responsible for the yellowleaf color in P2.These preliminary data are valuable for further exploring the molecular mechanisms of leaf color formation and associated pathways.展开更多
Rubus chingii Hu, a member of the rosaceae family, is extensively distributed in China and Japan. Its unripe fruits(Fupenzi in Chinese) have a long history of use as an herbal tonic in traditional Chinese medicine for...Rubus chingii Hu, a member of the rosaceae family, is extensively distributed in China and Japan. Its unripe fruits(Fupenzi in Chinese) have a long history of use as an herbal tonic in traditional Chinese medicine for treating various diseases commonly associated with kidney deficiency, and they are still in use today. Phytochemical investigations on the fruits and leaves of R. chingii indicate the presence of terpenoids, flavonoids, steroids, alkaloids, phenylpropanoids, phenolics, and organic acids. Extracts or active substances from this plant are reported to have various pharmacological properties, including antioxidant, anti-inflammatory, antitumor, antifungal, antithrombotic, antiosteoporotic, hypoglycemic, and central nervous system-regulating effects. This review provides up-to-date information on the botanical characterizations, traditional usages, chemical constituents, pharmacological activities, toxicity,and quality control of R. chingii. Possible directions for future research are also briefly proposed. This review aims to supply fundamental data for the further study of R. chingii and contribute to the development of its clinical use.展开更多
Protein tyrosine phosphatase 1 B(PTP1 B) has led to an intense interest in developing its inhibitors as anti-diabetes, anti-obesity and anti-cancer agents. The fruits of Rubus chingii(Chinese raspberry) were used as a...Protein tyrosine phosphatase 1 B(PTP1 B) has led to an intense interest in developing its inhibitors as anti-diabetes, anti-obesity and anti-cancer agents. The fruits of Rubus chingii(Chinese raspberry) were used as a kind of dietary traditional Chinese medicine. The methanolic extract of R. chingii fruits exhibited significant PTP1 B inhibitory activity. Further bioactivity-guided fractionation resulted in the isolation of three PTP1 B inhibitory ursane-type triterpenes: ursolic acid(1), 2-oxopomolic acid(2), and 2α, 19α-dihydroxy-3-oxo-urs-12-en-28-oic acid(3). Kinetics analyses revealed that 1 was a non-competitive PTP1 B inhibitor, and 2 and 3 were mixed type PTP1 B inhibitors. Compounds 1-3 and structurally related triterpenes(4-8) were further analyzed the structure-activity relationship, and were evaluated the inhibitory selectivity against four homologous protein tyrosine phosphatases(TCPTP, VHR, SHP-1 and SHP-2). Molecular docking simulations were also carried out, and the result indicated that 1, 3-acetoxyurs-12-ene-28-oic acid(5), and pomolic acid-3β-acetate(6) bound at the allosteric site including α3, α6, and α7 helix of PTP1 B.展开更多
Objective: In this study, one of the objectives was to investigate the total flavonoid contents of Fupenzi (R. chingii Hu.) obtained from different regions of China and to evaluate their anatioxidant activities. And t...Objective: In this study, one of the objectives was to investigate the total flavonoid contents of Fupenzi (R. chingii Hu.) obtained from different regions of China and to evaluate their anatioxidant activities. And the second objective of this study was to develop a validated HPLC method for chromatographic fingerprints of the samples extracts of Fupenzi. Method: The total flavonoid contents were determined by a colorimetric method and the antioxidant activity was determined spectrophotometrically by DPPH and ABTS radical scavenging assays. The chromatographic fingerprint was developed by high-performance liquid chromatography coupled with diode array detection for the control of Fupenzi. Results: A significant correlation between antioxidant activity and the total flavonoid content was observed for the DPPH assay (r2 = 0.758, ρ = 0.004) and the ABTS assay (r2 = 0.788, ρ = 0.002). Under the optimized chromatographic conditions, the validated method was successfully applied to assessment of chemical fingerprinting of 12 batches of FPZ collected from different regions of China. Comparisons of the chromatograms showed that 15 characteristic peaks could be selected as markers for identification and evaluation of Fupenzi. In addition, the proposed method was also successfully applied to simultaneous determination of five compounds (including puerarin, rutin, hyperin, quercetin and kaempferol) in these samples. Conclusions: The qualitative and quantitative analysis described in this paper could be used for identification and evaluation of Fupenzi.展开更多
Objective To develop a simple, efficient, and reliable method for routine quantitative analysis of main constituents presented in the fruits of Rubus chingii, which is widely used in Chinese materia medica （CMM）, kn...Objective To develop a simple, efficient, and reliable method for routine quantitative analysis of main constituents presented in the fruits of Rubus chingii, which is widely used in Chinese materia medica （CMM）, known as Fupenzi （FPZ） in Chinese. Methods An ultra performance liquid chromatography-photo diode array （UPLC-PDA） system was employed for simultaneous quantification of eight compounds, i. e. adenosine, gallic acid, brevifolin carboxylic acid, ethyl gallate, ellagic acid, kaempferol-3-O-rutinoside, kaempferol-3-O-^-D-glucopyranoside, and tiliroside. The chromatographic analysis was performed on a C18 column using a gradient elution of acetonitrile -0.1% formic acid aqueous solution within a runtime of 25 min. Results All calibration curves were linear （R2〉 0.9997） over the tested ranges. The intra- and inter-day precisions as determined from sample solutions were both less than 2.45% and 2.78%, respectively. The average recoveries for the eight constituents ranged from 94.77% to 101.35% with RSD ≤ 4.41%. The newly-developed method was applied to the quality assessment of various R. chingii samples, including both ripe and unripe fruits of R. ching/i from different habitats. Conclusion The relative levels of the investigated compounds vary remarkably in the fruits of R. chingiicollected from different habitats. As only two of the eight compounds, adenosine and ellagic acid, are determined in the ripe fruits of R. chingii, the results may explain the reason why only the unripe fruits can be used in CMM.展开更多
Raspberries are used for both food and medicine,but it has not yet attracted widespread attention.In this paper,the chemical constituen of the original plant raspberry.R.chingii is one of the new“Zhe Bawei”medicinal...Raspberries are used for both food and medicine,but it has not yet attracted widespread attention.In this paper,the chemical constituen of the original plant raspberry.R.chingii is one of the new“Zhe Bawei”medicinal materials selected in 2017.“Zhe Bawei”refers to eight kinds of genuine medicinal materials in Zhejiang Province.The chemical constituents,pharmacological effects,processing,and application of Rubus chingii Hu were reviewed to provide a reference for its further development.Relevant literature in recent years was collected in databases such as China Knowledge Network,Web of Science,Elsevier,PubMed,and X-Mol,using“raspberry”,“Rubus chingii”,“traditional use”,“chemical composition”,“pharmacology”,etc.as keywords individually or in combination.The summary of pharmacological activities shows that the relationship between the pharmacological activities of raspberry is still not deep enough.More in-depth research should be carried out in this direction to explore the mechanism of action of its active ingredients and provide effective reference for the further development of the raspberry industry.In the future,with the participation of more researchers,it is expected to develop innovative drugs based on raspberry for the treatment of diseases.展开更多
目的探究不同生长时期覆盆子Rubus chingii果实中的黄酮类物质的变化规律,解析其苷类物质积累分子机制。方法利用UPLC-MS/MS靶向代谢组学技术和高通量转录组测序(RNA sequencing,RNA-seq)分析了覆盆子果实5个不同生长时期S1(小青果)、S2...目的探究不同生长时期覆盆子Rubus chingii果实中的黄酮类物质的变化规律,解析其苷类物质积累分子机制。方法利用UPLC-MS/MS靶向代谢组学技术和高通量转录组测序(RNA sequencing,RNA-seq)分析了覆盆子果实5个不同生长时期S1(小青果)、S2(大青果)、S3(绿转黄果)、S4(黄转红果)、S5(红果)黄酮类化合物积累的变化,利用MetaboAnalyst结合KEGG、MWDB等数据库进行代谢物的功能注释,并利用主成分分析(principal component analysis,PCA)、正交偏最小二乘判别分析(orthogonal partial least squares-discriminant analysis,OPLS-DA)筛选差异代谢物,并将2组学进行联合分析。结果利用UPLC-MS/MS从5个不同时期覆盆子果实共鉴定出87种代谢物,其中分别检测出81(S1)、77(S2)、78(S3)、76(S4)、77(S5)种差异积累代谢物,差异代谢物主要富集在黄酮生物合成、异黄酮生物合成、黄酮和黄酮醇生物合成、次级代谢物的生物合成等途径。利用高通量测序对5个不同时期覆盆子果实进行转录组文库构建,共获得114.7 Gb的Clean Data,每个样品均达到6.23 M clean reads以上,各样本Q30碱基占92.3%及以上。将不同生长时期的转录组进行比对,差异基因表达分别为有2354个(S1 vs S2)、1027个(S1 vs S3)、3370个(S1 vs S4)、10249个(S1 vs S5)。结论从代谢和转录的2个层面上对覆盆子不同生长时期中的黄酮代谢物变化和转录变化展开分析,共注释到了黄酮合成途径的15个黄酮代谢物和95个基因,将其进行相关性分析发现,查耳酮异构酶(chalcone isomerase,CHI)、对羟基肉桂酰转移酶(hydroxycinnamoyl transferase,HCT)、查耳酮合酶(chalcone synthase,CHS)、黄酮醇合酶(flavonol synthase,FLS)、黄烷酮-3-羟化酶(flavanone 3-hydroxylase,F3H)、黄酮3’-羟化酶(flavonoid 3'-hydroxylase,F3′H)等基因的表达水平与黄酮代谢物的积累显著相关,表明这些基因参与调控黄酮类化合物的生物合成。首次阐述了覆盆子的黄酮类成分的合成途径和相关基因,为利用生物工程技术生产其黄酮类化合物提供了依据,对于扩大药用资源具有重要意义。展开更多
基金funded by the Beijing Gardening andGreeningYouth InnovationTalent Support Program(kjcx202336)theKey R&D Project of theOpen Subject of the Beijing Key Laboratory for Greening Plant Breeding(YZZD202403).
文摘Naturally occurring yellow leaf mutants are an important resource for studying pigment content and biosynthesis,as well as related gene expression.In our ongoing cultivation of Rehmannia chingii H.L.Li,we found an off-type yellow plant.The yellowing started with the new leaves and gradually spread downward until the entire plant exhibited a stable shade of yellow.We studied the differences in the chlorophyll and carotenoid content,carotenoid profile,and transcriptome of this yellow-leaf mutant(P2).Compared to the wild-type R.chingii plant(P1),P2 leaves had significantly lower chlorophyll and carotenoid content.LC-MS/MS analysis revealed that P2 had higher quantities of severalmetabolites in the carotenoid biosynthesis pathway.Transcriptome sequencing results showed that genes involved in porphyrin metabolism,carbon fixation,photosynthesis and antenna proteins,terpenoid backbone biosynthesis,and carotenoid biosynthesis were differentially expressed between P1 and P2.Large-scale expression differences were observed in the phytohormone and MAPK signaling pathways,as well as in 15 transcription factor families.We discuss possible mechanisms responsible for the yellowleaf color in P2.These preliminary data are valuable for further exploring the molecular mechanisms of leaf color formation and associated pathways.
基金the National Natural Science Foundation of China(No.81803794)the Program for Excellent Talents in Zhejiang Chinese Medical University(No.112123A12201/001/004/019)。
文摘Rubus chingii Hu, a member of the rosaceae family, is extensively distributed in China and Japan. Its unripe fruits(Fupenzi in Chinese) have a long history of use as an herbal tonic in traditional Chinese medicine for treating various diseases commonly associated with kidney deficiency, and they are still in use today. Phytochemical investigations on the fruits and leaves of R. chingii indicate the presence of terpenoids, flavonoids, steroids, alkaloids, phenylpropanoids, phenolics, and organic acids. Extracts or active substances from this plant are reported to have various pharmacological properties, including antioxidant, anti-inflammatory, antitumor, antifungal, antithrombotic, antiosteoporotic, hypoglycemic, and central nervous system-regulating effects. This review provides up-to-date information on the botanical characterizations, traditional usages, chemical constituents, pharmacological activities, toxicity,and quality control of R. chingii. Possible directions for future research are also briefly proposed. This review aims to supply fundamental data for the further study of R. chingii and contribute to the development of its clinical use.
基金supported by the National Natural Science Foundation of China(No.81628012)
文摘Protein tyrosine phosphatase 1 B(PTP1 B) has led to an intense interest in developing its inhibitors as anti-diabetes, anti-obesity and anti-cancer agents. The fruits of Rubus chingii(Chinese raspberry) were used as a kind of dietary traditional Chinese medicine. The methanolic extract of R. chingii fruits exhibited significant PTP1 B inhibitory activity. Further bioactivity-guided fractionation resulted in the isolation of three PTP1 B inhibitory ursane-type triterpenes: ursolic acid(1), 2-oxopomolic acid(2), and 2α, 19α-dihydroxy-3-oxo-urs-12-en-28-oic acid(3). Kinetics analyses revealed that 1 was a non-competitive PTP1 B inhibitor, and 2 and 3 were mixed type PTP1 B inhibitors. Compounds 1-3 and structurally related triterpenes(4-8) were further analyzed the structure-activity relationship, and were evaluated the inhibitory selectivity against four homologous protein tyrosine phosphatases(TCPTP, VHR, SHP-1 and SHP-2). Molecular docking simulations were also carried out, and the result indicated that 1, 3-acetoxyurs-12-ene-28-oic acid(5), and pomolic acid-3β-acetate(6) bound at the allosteric site including α3, α6, and α7 helix of PTP1 B.
文摘Objective: In this study, one of the objectives was to investigate the total flavonoid contents of Fupenzi (R. chingii Hu.) obtained from different regions of China and to evaluate their anatioxidant activities. And the second objective of this study was to develop a validated HPLC method for chromatographic fingerprints of the samples extracts of Fupenzi. Method: The total flavonoid contents were determined by a colorimetric method and the antioxidant activity was determined spectrophotometrically by DPPH and ABTS radical scavenging assays. The chromatographic fingerprint was developed by high-performance liquid chromatography coupled with diode array detection for the control of Fupenzi. Results: A significant correlation between antioxidant activity and the total flavonoid content was observed for the DPPH assay (r2 = 0.758, ρ = 0.004) and the ABTS assay (r2 = 0.788, ρ = 0.002). Under the optimized chromatographic conditions, the validated method was successfully applied to assessment of chemical fingerprinting of 12 batches of FPZ collected from different regions of China. Comparisons of the chromatograms showed that 15 characteristic peaks could be selected as markers for identification and evaluation of Fupenzi. In addition, the proposed method was also successfully applied to simultaneous determination of five compounds (including puerarin, rutin, hyperin, quercetin and kaempferol) in these samples. Conclusions: The qualitative and quantitative analysis described in this paper could be used for identification and evaluation of Fupenzi.
基金National Science and Technology Major Project for"Major New Drugs Innovation and Development"(2015ZX09J15102-004-004)
文摘Objective To develop a simple, efficient, and reliable method for routine quantitative analysis of main constituents presented in the fruits of Rubus chingii, which is widely used in Chinese materia medica （CMM）, known as Fupenzi （FPZ） in Chinese. Methods An ultra performance liquid chromatography-photo diode array （UPLC-PDA） system was employed for simultaneous quantification of eight compounds, i. e. adenosine, gallic acid, brevifolin carboxylic acid, ethyl gallate, ellagic acid, kaempferol-3-O-rutinoside, kaempferol-3-O-^-D-glucopyranoside, and tiliroside. The chromatographic analysis was performed on a C18 column using a gradient elution of acetonitrile -0.1% formic acid aqueous solution within a runtime of 25 min. Results All calibration curves were linear （R2〉 0.9997） over the tested ranges. The intra- and inter-day precisions as determined from sample solutions were both less than 2.45% and 2.78%, respectively. The average recoveries for the eight constituents ranged from 94.77% to 101.35% with RSD ≤ 4.41%. The newly-developed method was applied to the quality assessment of various R. chingii samples, including both ripe and unripe fruits of R. ching/i from different habitats. Conclusion The relative levels of the investigated compounds vary remarkably in the fruits of R. chingiicollected from different habitats. As only two of the eight compounds, adenosine and ellagic acid, are determined in the ripe fruits of R. chingii, the results may explain the reason why only the unripe fruits can be used in CMM.
基金supported by the Innovation Team and Talents Cultivation Program of National Administration of Traditional Chinese Medicine(No.ZYYCXTD-D-202005)Tianjin Municipal Science and Technology Committee(No.20YFZCSY00560)+1 种基金the Key Project at Central Government Level(No.2060302)National Natural Science Foundation of China(No.82373982,82173969).
文摘Raspberries are used for both food and medicine,but it has not yet attracted widespread attention.In this paper,the chemical constituen of the original plant raspberry.R.chingii is one of the new“Zhe Bawei”medicinal materials selected in 2017.“Zhe Bawei”refers to eight kinds of genuine medicinal materials in Zhejiang Province.The chemical constituents,pharmacological effects,processing,and application of Rubus chingii Hu were reviewed to provide a reference for its further development.Relevant literature in recent years was collected in databases such as China Knowledge Network,Web of Science,Elsevier,PubMed,and X-Mol,using“raspberry”,“Rubus chingii”,“traditional use”,“chemical composition”,“pharmacology”,etc.as keywords individually or in combination.The summary of pharmacological activities shows that the relationship between the pharmacological activities of raspberry is still not deep enough.More in-depth research should be carried out in this direction to explore the mechanism of action of its active ingredients and provide effective reference for the further development of the raspberry industry.In the future,with the participation of more researchers,it is expected to develop innovative drugs based on raspberry for the treatment of diseases.
文摘目的探究不同生长时期覆盆子Rubus chingii果实中的黄酮类物质的变化规律,解析其苷类物质积累分子机制。方法利用UPLC-MS/MS靶向代谢组学技术和高通量转录组测序(RNA sequencing,RNA-seq)分析了覆盆子果实5个不同生长时期S1(小青果)、S2(大青果)、S3(绿转黄果)、S4(黄转红果)、S5(红果)黄酮类化合物积累的变化,利用MetaboAnalyst结合KEGG、MWDB等数据库进行代谢物的功能注释,并利用主成分分析(principal component analysis,PCA)、正交偏最小二乘判别分析(orthogonal partial least squares-discriminant analysis,OPLS-DA)筛选差异代谢物,并将2组学进行联合分析。结果利用UPLC-MS/MS从5个不同时期覆盆子果实共鉴定出87种代谢物,其中分别检测出81(S1)、77(S2)、78(S3)、76(S4)、77(S5)种差异积累代谢物,差异代谢物主要富集在黄酮生物合成、异黄酮生物合成、黄酮和黄酮醇生物合成、次级代谢物的生物合成等途径。利用高通量测序对5个不同时期覆盆子果实进行转录组文库构建,共获得114.7 Gb的Clean Data,每个样品均达到6.23 M clean reads以上,各样本Q30碱基占92.3%及以上。将不同生长时期的转录组进行比对,差异基因表达分别为有2354个(S1 vs S2)、1027个(S1 vs S3)、3370个(S1 vs S4)、10249个(S1 vs S5)。结论从代谢和转录的2个层面上对覆盆子不同生长时期中的黄酮代谢物变化和转录变化展开分析,共注释到了黄酮合成途径的15个黄酮代谢物和95个基因,将其进行相关性分析发现,查耳酮异构酶(chalcone isomerase,CHI)、对羟基肉桂酰转移酶(hydroxycinnamoyl transferase,HCT)、查耳酮合酶(chalcone synthase,CHS)、黄酮醇合酶(flavonol synthase,FLS)、黄烷酮-3-羟化酶(flavanone 3-hydroxylase,F3H)、黄酮3’-羟化酶(flavonoid 3'-hydroxylase,F3′H)等基因的表达水平与黄酮代谢物的积累显著相关,表明这些基因参与调控黄酮类化合物的生物合成。首次阐述了覆盆子的黄酮类成分的合成途径和相关基因,为利用生物工程技术生产其黄酮类化合物提供了依据,对于扩大药用资源具有重要意义。
