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Abstract Background During Fritillaria thunbergii planting, pests and diseases usually invade the plant, resulting in reduced yield and quality. Previous studies have demonstrated that using biocontrol agents can effectively control grubs and affect the steroid alkaloids content in F. thunbergii. However, the molecular regulatory mechanisms underlying the differences in the accumulation of steroid alkaloids in response to biocontrol agents remain unclear. Results Combined transcriptomic and metabolic analyses were performed by treating the bulbs of F. thunbergii treated with biocontrol agents during planting. Otherwise, 48 alkaloids including 32 steroid alkaloids, 6 indole alkaloids, 2 scopolamine-type alkaloids, 1 isoquinoline alkaloid, 1 furoquinoline alkaloid, and 6 other alkaloids were identified. The content of steroidal alkaloids particularly peimine, peiminine, and veratramine, increased significantly in the group treated with the biocontrol agents. Transcriptome sequencing identified 929 differential genes using biocontrol agents, including 589 upregulated and 340 downregulated genes. Putative biosynthesis networks of steroid alkaloids have been established and combined with differentially expressed structural unigenes, such as acetyl-CoA C-acetyl-transferase, acelyl-CoAC-acetyltransferase3-hydroxy-3-methylglutaryl-coenzyme A synthase, 1-deoxy-D-xylulose-5-phosphate reductor-isomerase, 2-C-methyl-D-erythritol-4-phosphate cytidylyltransferase and 4-hydroxy-3-methylbut-2-enyl diphosphate reductase. In addition, biological processes such as amino acid accumulation and oxidative phosphorylation were predicted to be related to the synthesis of steroid alkaloids. Cytochrome P450 enzymes also play crucial roles in the steroid alkaloid synthesis. The transcription factor families MYB and bHLH were significantly upregulated after using biocontrol agents. Conclusions Biocontrol agents increased the steroid alkaloids accumulation of steroid alkaloids by affecting key enzymes in the steroid alkaloid synthesis pathway, biological processes of oxidative phosphorylation and amino acid synthesis, cytochrome P450 enzymes, and transcription factors. This study revealed the mechanism underlying the difference in steroidal alkaloids in F. thunbergii after using biocontrol agents, laying the groundwork for future industrial production of steroid alkaloids and ecological planting of medicinal materials in the future.

The continuous obstacles of cropping cause severe economic loss, which seriously threaten agricultural sustainable development. In addition, managing excess waste, such as potato peel and mineral waste residues, is a vital burden for industry and agriculture. Therefore, we explored the feasibility of reductive soil disinfestation (RSD) with potato peel and amendment with iron mineral waste residues for the production of Fritillaria thunbergii, which is vulnerable to continuous obstacles. In this study, the influences of iron mineral, RSD with different organic maters, as well as the combined effects of iron mineral and RSD on Fritillaria rhizosphere soil physicochemical properties, microbial communities, and Fritillaria production were investigated. The results revealed that the RSD treatments with potato peel significantly reduced the soil salinity and increased the soil pH, microbial activity, organic matter, and the contents of K and Ca. RSD with potato peel also significantly thrived of the beneficial microbes (Bacillus, Azotobacter, Microvirga, and Chaetomium), and down-regulated potential plant pathogens. RSD with potato peel significantly promoted F. thunbergii yield and quality. Moreover, the combined effects of RSD and iron mineral amendment further enhanced soil health, improved microbial community composition, and increased the yield and peimisine content of F. thunbergii by 24.2% and 49.3%, respectively. Overall, our results demonstrated that RSD with potato peel and amendment with iron mineral waste residues can efficiently improve soil fertility, modify the microbial community, and benefit for both the sustainable production of F. thunbergii and the management of waste. Key points: • RSD increases soil pH, organic matter, microbial activity, and mineral content • RSD with potato peel enriches beneficial microbes and decreases plant pathogens • PP + Fe treatment increases Fritillaria yield by 24.2% and peimisine content by 49.3% [ABSTRACT FROM AUTHOR]

Background: During Fritillaria thunbergii planting, pests and diseases usually invade the plant, resulting in reduced yield and quality. Previous studies have demonstrated that using biocontrol agents can effectively control grubs and affect the steroid alkaloids content in F. thunbergii. However, the molecular regulatory mechanisms underlying the differences in the accumulation of steroid alkaloids in response to biocontrol agents remain unclear. Results: Combined transcriptomic and metabolic analyses were performed by treating the bulbs of F. thunbergii treated with biocontrol agents during planting. Otherwise, 48 alkaloids including 32 steroid alkaloids, 6 indole alkaloids, 2 scopolamine-type alkaloids, 1 isoquinoline alkaloid, 1 furoquinoline alkaloid, and 6 other alkaloids were identified. The content of steroidal alkaloids particularly peimine, peiminine, and veratramine, increased significantly in the group treated with the biocontrol agents. Transcriptome sequencing identified 929 differential genes using biocontrol agents, including 589 upregulated and 340 downregulated genes. Putative biosynthesis networks of steroid alkaloids have been established and combined with differentially expressed structural unigenes, such as acetyl-CoA C-acetyl-transferase, acelyl-CoAC-acetyltransferase3-hydroxy-3-methylglutaryl-coenzyme A synthase, 1-deoxy-D-xylulose-5-phosphate reductor-isomerase, 2-C-methyl-D-erythritol-4-phosphate cytidylyltransferase and 4-hydroxy-3-methylbut-2-enyl diphosphate reductase. In addition, biological processes such as amino acid accumulation and oxidative phosphorylation were predicted to be related to the synthesis of steroid alkaloids. Cytochrome P450 enzymes also play crucial roles in the steroid alkaloid synthesis. The transcription factor families MYB and bHLH were significantly upregulated after using biocontrol agents. Conclusions: Biocontrol agents increased the steroid alkaloids accumulation of steroid alkaloids by affecting key enzymes in the steroid alkaloid synthesis pathway, biological processes of oxidative phosphorylation and amino acid synthesis, cytochrome P450 enzymes, and transcription factors. This study revealed the mechanism underlying the difference in steroidal alkaloids in F. thunbergii after using biocontrol agents, laying the groundwork for future industrial production of steroid alkaloids and ecological planting of medicinal materials in the future. [ABSTRACT FROM AUTHOR]

In this paper, the samples of Fritillaria hupehensis and Fritillaria thunbergii were collected from the same area with identical growth cycle, and their compositions of TPC (total phenolics content), TFC (total flavonoids content), free amino acids, trace elements, nucleosides and nucleobases were comprehensively compared. Besides that, their corresponding physicochemical properties of antioxidant ability, solubility, swelling and gelatinization characteristic were also discussed. Compared with Fritillaria hupehensis, the Fritillaria thunbergii showed higher antioxidant activities in mature bulb due to its higher TPC, TFC, DPPH, FRAP and ABTS values. On the other hand, the contents of total nucleosides, nucleobases, amino acids in bulb of Fritillaria hupehensis were significantly higher than those of Fritillaria thunbergii (P

Fast detection of heavy metals is important to ensure the quality and safety of herbal medicines. In this study, laser-induced breakdown spectroscopy (LIBS) was applied to detect the heavy metal content (Cd, Cu, and Pb) in Fritillaria thunbergii. Quantitative prediction models were established using a back-propagation neural network (BPNN) optimized using the particle swarm optimization (PSO) algorithm and sparrow search algorithm (SSA), called PSO-BP and SSA-BP, respectively. The results revealed that the BPNN models optimized by PSO and SSA had better accuracy than the BPNN model without optimization. The performance evaluation metrics of the PSO-BP and SSA-BP models were similar. However, the SSA-BP model had two advantages: it was faster and had higher prediction accuracy at low concentrations. For the three heavy metals Cd, Cu and Pb, the prediction correlation coefficient (Rp2) values for the SSA-BP model were 0.972, 0.991 and 0.956; the prediction root mean square error (RMSEP) values were 5.553, 7.810 and 12.906 mg/kg; and the prediction relative percent deviation (RPD) values were 6.04, 10.34 and 4.94, respectively. Therefore, LIBS could be considered a constructive tool for the quantification of Cd, Cu and Pb contents in Fritillaria thunbergii. [ABSTRACT FROM AUTHOR]

Environmental and health risks associated with heavy metal pollution are serious. Human health can be adversely affected by the smallest amount of heavy metals. Modeling spectrum requires the careful selection of variables. Hence, simple variables that have a low level of interference and a high degree of precision are required for fast analysis and online detection. This study used laser-induced breakdown spectroscopy coupled with variable selection and chemometrics to simultaneously analyze heavy metals (Cd, Cu and Pb) in Fritillaria thunbergii. A total of three machine learning algorithms were utilized, including a gradient boosting machine (GBM), partial least squares regression (PLSR) and support vector regression (SVR). Three promising wavelength selection methods were evaluated for comparison, namely, a competitive adaptive reweighted sampling method (CARS), a random frog method (RF), and an uninformative variable elimination method (UVE). Compared to full wavelengths, the selected wavelengths produced excellent results. Overall, RC2, RV2, RP2, RSMEC, RSMEV and RSMEP for the selected variables are as follows: 0.9967, 0.8899, 0.9403, 1.9853 mg kg−1, 11.3934 mg kg−1, 8.5354 mg kg−1; 0.9933, 0.9316, 0.9665, 5.9332 mg kg−1, 18.3779 mg kg−1, 11.9356 mg kg−1; 0.9992, 0.9736, 0.9686, 1.6707 mg kg−1, 10.2323 mg kg−1, 10.1224 mg kg−1 were obtained for Cd Cu and Pb, respectively. Experimental results showed that all three methods could perform variable selection effectively, with GBM-UVE for Cd, SVR-RF for Pb, and GBM-CARS for Cu providing the best results. The results of the study suggest that LIBS coupled with wavelength selection can be used to detect heavy metals rapidly and accurately in Fritillaria by extracting only a few variables that contain useful information and eliminating non-informative variables. [ABSTRACT FROM AUTHOR]

Both the bulbs and flowers of Fritillaria thunbergii Miq. (BFT and FFT) are widely applied as expectorants and antitussives in traditional Chinese medicine, but few studies have been conducted to compare the chemical compositions of these plant parts. In this study, 50% methanol extracts of BFT and FFT were analyzed via UHPLC-Q-Exactive Orbitrap MS/MS, and the feasibility of using non-targeted UHPLC-HRMS metabolomics and molecular networking to address the authentication of bulb and flower samples was evaluated. Principal component analysis (PCA), Orthogonal Partial Least Squares-Discriminant Analysis (OPLS-DA), and heat map analysis showed there were dissimilar metabolites in BFT and FFT. As a result, 252 and 107 peaks in positive ion mode and negative mode, respectively, were considered to represent significant difference variables between BFT and FFT. Then, MS/MS-based molecular networking of BFT and FFT was constructed to perform an in-depth characterization of the peaks using different variables. A total of 31 alkaloids with significant differences were annotated in this paper, including seven cis-D/E-vevanine without C20-OH and one trans-D/E-cevanine with C20-OH, thirteen trans-D/E-cevanine without C20-OH, five cevanine N-oxide, and five veratramine. Among the 31 alkaloids, eight alkaloids had higher FFT than BFT contents, while all the flavonoids identified in our work had greater FFT than BFT contents. The influence of different ingredients on the pharmacological activities of BFT and FFT should be investigated in future studies. [ABSTRACT FROM AUTHOR]

Traditional Chinese herbal medicine (TCHM) plays an essential role in the international pharmaceutical industry due to its rich resources and unique curative properties. The flowers, stems, and leaves of Fritillaria contain a wide range of phytochemical compounds, including flavonoids, essential oils, saponins, and alkaloids, which may be useful for medicinal purposes. Fritillaria thunbergii Miq. Bulbs are commonly used in traditional Chinese medicine as expectorants and antitussives. In this paper, a feasibility study is presented that examines the use of hyperspectral imaging integrated with convolutional neural networks (CNN) to distinguish twelve (12) Fritillaria varieties (n = 360). The performance of support vector machines (SVM) and partial least squares-discriminant analysis (PLS-DA) was compared with that of convolutional neural network (CNN). Principal component analysis (PCA) was used to assess the presence of cluster trends in the spectral data. To optimize the performance of the models, cross-validation was used. Among all the discriminant models, CNN was the most accurate with 98.88%, 88.89% in training and test sets, followed by PLS-DA and SVM with 92.59%, 81.94% and 99.65%, 79.17%, respectively. The results obtained in the present study revealed that application of HSI in conjunction with the deep learning technique can be used for classification of Fritillaria thunbergii varieties rapidly and non-destructively. [ABSTRACT FROM AUTHOR]

Fast detection of heavy metals is important to ensure the quality and safety of herbal medicines. In this study, laser-induced breakdown spectroscopy (LIBS) was applied to detect the heavy metal content (Cd, Cu, and Pb) in Fritillaria thunbergii. Quantitative prediction models were established using a back-propagation neural network (BPNN) optimized using the particle swarm optimization (PSO) algorithm and sparrow search algorithm (SSA), called PSO-BP and SSA-BP, respectively. The results revealed that the BPNN models optimized by PSO and SSA had better accuracy than the BPNN model without optimization. The performance evaluation metrics of the PSO-BP and SSA-BP models were similar. However, the SSA-BP model had two advantages: it was faster and had higher prediction accuracy at low concentrations. For the three heavy metals Cd, Cu and Pb, the prediction correlation coefficient (Rp2) values for the SSA-BP model were 0.972, 0.991 and 0.956; the prediction root mean square error (RMSEP) values were 5.553, 7.810 and 12.906 mg/kg; and the prediction relative percent deviation (RPD) values were 6.04, 10.34 and 4.94, respectively. Therefore, LIBS could be considered a constructive tool for the quantification of Cd, Cu and Pb contents in Fritillaria thunbergii.

Environmental and health risks associated with heavy metal pollution are serious. Human health can be adversely affected by the smallest amount of heavy metals. Modeling spectrum requires the careful selection of variables. Hence, simple variables that have a low level of interference and a high degree of precision are required for fast analysis and online detection. This study used laser-induced breakdown spectroscopy coupled with variable selection and chemometrics to simultaneously analyze heavy metals (Cd, Cu and Pb) in Fritillaria thunbergii. A total of three machine learning algorithms were utilized, including a gradient boosting machine (GBM), partial least squares regression (PLSR) and support vector regression (SVR). Three promising wavelength selection methods were evaluated for comparison, namely, a competitive adaptive reweighted sampling method (CARS), a random frog method (RF), and an uninformative variable elimination method (UVE). Compared to full wavelengths, the selected wavelengths produced excellent results. Overall, RC2, RV2, RP2, RSMEC, RSMEV and RSMEP for the selected variables are as follows: 0.9967, 0.8899, 0.9403, 1.9853 mg kg−1, 11.3934 mg kg−1, 8.5354 mg kg−1; 0.9933, 0.9316, 0.9665, 5.9332 mg kg−1, 18.3779 mg kg−1, 11.9356 mg kg−1; 0.9992, 0.9736, 0.9686, 1.6707 mg kg−1, 10.2323 mg kg−1, 10.1224 mg kg−1 were obtained for Cd Cu and Pb, respectively. Experimental results showed that all three methods could perform variable selection effectively, with GBM-UVE for Cd, SVR-RF for Pb, and GBM-CARS for Cu providing the best results. The results of the study suggest that LIBS coupled with wavelength selection can be used to detect heavy metals rapidly and accurately in Fritillaria by extracting only a few variables that contain useful information and eliminating non-informative variables.

Both the bulbs and flowers of Fritillaria thunbergii Miq. (BFT and FFT) are widely applied as expectorants and antitussives in traditional Chinese medicine, but few studies have been conducted to compare the chemical compositions of these plant parts. In this study, 50% methanol extracts of BFT and FFT were analyzed via UHPLC-Q-Exactive Orbitrap MS/MS, and the feasibility of using non-targeted UHPLC-HRMS metabolomics and molecular networking to address the authentication of bulb and flower samples was evaluated. Principal component analysis (PCA), Orthogonal Partial Least Squares-Discriminant Analysis (OPLS-DA), and heat map analysis showed there were dissimilar metabolites in BFT and FFT. As a result, 252 and 107 peaks in positive ion mode and negative mode, respectively, were considered to represent significant difference variables between BFT and FFT. Then, MS/MS-based molecular networking of BFT and FFT was constructed to perform an in-depth characterization of the peaks using different variables. A total of 31 alkaloids with significant differences were annotated in this paper, including seven cis-D/E-vevanine without C20-OH and one trans-D/E-cevanine with C20-OH, thirteen trans-D/E-cevanine without C20-OH, five cevanine N-oxide, and five veratramine. Among the 31 alkaloids, eight alkaloids had higher FFT than BFT contents, while all the flavonoids identified in our work had greater FFT than BFT contents. The influence of different ingredients on the pharmacological activities of BFT and FFT should be investigated in future studies.

In order to explore the relationship bewteen the accumulation of medicinal components and the expression level of alkaloid synthesis related genes in Fritillaria thunbergii, ultra high performance liquid chromotography mass spectrometry (UPLC-MS) and real-time quantitative PCR(qPCR) techniques were used to determined the total alkaloids (the sum of peimine and peiminine) and the expression level of three genes involved in alkaloids synthesis pathway (HMGR, FPS and DXR) in eleven samples from different producing areas. Meanwhile, biostatistics was used to analyze the correlation between the alkaloids content of mature bulbs and the expression of each gene. The results were as follows: There were significant differences in total alkaloid content of mature bulbs (P< 0.05), ranging from 0.2105% to 0.4612%; The variation trend of the expression of HMGR and FPS genes in the tissue of full-bloom stage and the bulbs from full-bloom stage to maturity stage was basically consistent with the variation trend of alkaloid content; The expression of DXR gene was the highest in mature bulbs, and the change trend of the tissue expression at the full-bloom stage and the bulb expression from the full-bloom stage to maturity stage was generally inconsistent with the change trend of alkaloid content; The expression levels of HMGR and FPS genes were significantly or extremely significantly positively correlated with peimine, peiminine and total alkaloid content, (P<0.05 or P<0.001), and the coefficients between FPS gene expression and alkaloid content were the highest, which were 0.672, 0.631, 0.664, respectively, meanwhile, the coefficients between DXR gene expression and alkaloid content were low. In summary, the data indicate the accumulation of alkaloids is significantly regulated or modified by HMGR and FPS genes in the MVA pathway, but not by DXR genes in the MEP pathway. [ABSTRACT FROM AUTHOR]

A method for the simultaneous determination of imidacloprid, pyrisoxazole, fluxapyroxad, cyprodinil, pyraclostrobin, abamectin in Fritillaria thunbergii Miq. was developed using QuEChERS-ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). F. thunbergii samples were extracted by V(acetonitrile):V(water)=4:1. Then they were cleaned up by PSA 50 mg + C18 50 mg + MgSO4150 mg adsorbents and detected by UPLC-MS/MS under positive polarity multiple reactions monitoring (MRM) mode. The matrix-matched external standard calibration curves were used for quantitative analysis. The linearities of 6 pesticides were all in the range of 0.001-0.2 mg/L, with correlation coefficients higher than 0.994. The limit of quantification (LOQ) was 0.01 mg/kg and the limits of detection (LOD) of this new method were 2×10-5 mg/kg to 3×10-4 mg/kg in fresh F. thunbergii. While the LOQ was 0.05 mg/kg and the LOD of the proposed method were 1×10-4 mg/kg to 1×10-3 mg/kg in dry F. thunbergii. Recoveries at three spiked levels of 0.01, 0.5 and 2 mg/kg ranged from 80% to 109% with relative standard deviations (RSDs) from 0.95% to 13% (n=5) in fresh F. thunbergii. While, in dry F. thunbergii, recoveries at three spiked levels of 0.05, 0.5 and 2 mg/kg ranged from 77% to 101% with RSDs from 0.89% to 7.5% (n=5). The application of this new method in real samples has been demonstrated. Given that this method is fast, simple, accurate and efficient, it can meet the requirements for the determination of residues of imidacloprid, pyrisoxazole, fluxapyroxad, cyprodinil, pyraclostrobine, abamectin in F. thunbergii. [ABSTRACT FROM AUTHOR]

To explore rhizosphere effects, geographical differences and their effects on the bacterial community associated with the geoherb Fritillaria thunbergii, some physicochemical properties of soil samples (3 sampling sites × 2 habitats (rhizosphere and bulk soil)) were measured and the soil bacterial community detected by PCR-denaturing gradient gel electrophoresis (DGGE). Among the three regions, soil pH varied between 4.48 and 7.73 indicating that F. thunbergii could grow both in acid and slightly alkaline soil. As the authentic Dao-di producing area, Ningbo showed the highest soil quality with the highest content of organic matter (OM) (2.46%), phosphatase (268 mg kg−1 24 h−1) and urease activity (1481 mg kg−1 24 h−1). In comparison with the bulk soil, pH, organic carbon content, and phosphatase and urease activities were all lower in the rhizosphere, suggesting that the roots may secrete some unique metabolites in root exudates. Statistical analyses showed that soil properties of Ningbo and Panan in Zhejiang province were more similar to each other than those in Nantong in Jiangsu province. In addition, PCR-DGGE analysis showed that main bacterial population identified in F. thunbergii was proteobacteria (18 bands, 55%), acidobacteria (4, 12%), actinobacteria (4, 12%) and bacterioidetes (6, 18%). Overall, soil properties and microbial communities varied not only between the rhizosphere and bulk soil but also among the three regions. We suggest that the plant, together with the soil properties, cooperatively shape the structure of the rhizosphere bacteria, and that the soil properties have a close relationship with the geoherbalism of F. thunbergii.

Objective To observe the effects of the ethanol extract of thunberg fritillary leaf (EETFL) and the ethanol extract of thunberg fritillary flower (EETFF) on relieving cough, sputum elimination and relieving asthma. Methods The cough relieving effects of EETFL and EETFF were studied in mouse cough model caused by ammonia water and in guinea pig cough model caused by citric acid. The sputum elimination effects of EETFL and EETFF were researched by the observation of tracheal phenol red shedding in mice. The asthma relieving effects were tested by spraying method in guinea pigs. Results EETFL can obviously inhibit the incubation period and cough frequency of the model mice and guinea pigs induced by ammonia water and citric acid (P<0.05), and significantly improve the tracheal phenol red excretion volume in mice (P<0.05), and obviously prolong the incubation period of asthma (P<0.05). EETFF can obviously inhibit the incubation period and cough frequency of the model mice and guinea pigs induced by ammonia water and citric acid (P<0.05), and significantly improve the tracheal phenol red excretion volume in mice (P<0.05), but EETFF couldn't prolong the incubation period of asthma evidently. Conclusion EETFL has obvious activity of relieving cough, eliminating phlegm and relieving asthma. EETFF has obvious activity of relieving cough and eliminating phlegm, but EETFF has no anti-asthmatic activity under the current dose. [ABSTRACT FROM AUTHOR]

A method has been developed for the determination of pyrimethanil residue in Corydalis yanhusuo W. T. Wang, Fritillaria thunbergii Miq. and Paeonia lactiflora Pall. by using gas chromatography. Samples were extracted with dichloromethane, cleaned up by column chromatography (neutral alumina-florisil = 3 : 2, m/m) followed by liquid-liquid partition. Then pesticide residue was determined by gas chromatography with nitrogen-phosphorus detection (NPD), and the matrix-matched external standard method was applied for the quantitative analysis. The confirmation of pyrimethanil was performed by GC-MS/MS. Good linearity was obtained at concentrations of 0.02-10 mg/L with R2 higher than 0.999 in n-hexane and blank matrix solution. The spiked recoveries varied from 75% to 104% at 0.01 (0.02 in C. yanhusuo), 0.1, and 0.5 mg/kg spiked levels, respectively. The intra-day relative standard deviation ranged from 1.3% to 10.2% (n = 5) and the inter-day relative standard deviation ranged from 1.2% to 9.0% (n = 3), in various matrixes. The limit of quantification (LOQ) in C. yanhusuo, F. thunbergii and P. lactiflora was 0.02, 0.01, and 0.01 mg/kg, respectively. This analytical method met with the requirement of pesticide residue analysis. It was simple, reliable, and with better purification effect, which could be applied to the determination pyrimethanil residue in these three kinds of Chinese medicinal herbs. [ABSTRACT FROM AUTHOR]

Objective To set up the quality standard for Danggui Beimu Kushen Granules. Methods TLC was applied to qualitatively identify Angelica sinensis, Fritillaria thunbergⅡ, Sophora flavescens and HPLC was applied to quantitatively determine ferulic acid and matrine in the preparation. Results The TLC identification method of Angelica sinensis, Fritillaria ThunbergⅡand Sophora flavescens had strong specificity and good reproducibility; ferulic acid showed good linear relation in the range of 0.031 6~0.129 6μg (r=0. 999 8). The average recovery rate of ferulic acid was 101.620%, and RSD was 1.070%;matrine showed good linearity with the peak area in the range of 0.100 0~1.000 0 μg (r=0.999 6). The average recovery rate was 99.180%and RSD was 2.04%. Conclusion This methods are simple, sensitive and accurate with good reproducibility, and can be used as the quality control standard of Danggui Beimu Kushen Granules. [ABSTRACT FROM AUTHOR]

To explore rhizosphere effects, geographical differences and their effects on the bacterial community associated with the geoherb Fritillaria thunbergii, some physicochemical properties of soil samples (3 sampling sites × 2 habitats (rhizosphere and bulk soil)) were measured and the soil bacterial community detected by PCR-denaturing gradient gel electrophoresis (DGGE). Among the three regions, soil pH varied between 4.48 and 7.73 indicating that F. thunbergii could grow both in acid and slightly alkaline soil. As the authentic Dao-di producing area, Ningbo showed the highest soil quality with the highest content of organic matter (OM) (2.46%), phosphatase (268 mg kg-1 24 h-1) and urease activity (1481 mg kg-1 24 h-1). In comparison with the bulk soil, pH, organic carbon content, and phosphatase and urease activities were all lower in the rhizosphere, suggesting that the roots may secrete some unique metabolites in root exudates. Statistical analyses showed that soil properties of Ningbo and Panan in Zhejiang province were more similar to each other than those in Nantong in Jiangsu province. In addition, PCR-DGGE analysis showed that main bacterial population identified in F. thunbergii was proteobacteria (18 bands, 55%), acidobacteria (4, 12%), actinobacteria (4, 12%) and bacterioidetes (6, 18%). Overall, soil properties and microbial communities varied not only between the rhizosphere and bulk soil but also among the three regions. We suggest that the plant, together with the soil properties, cooperatively shape the structure of the rhizosphere bacteria, and that the soil properties have a close relationship with the geoherbalism of F. thunbergii. [ABSTRACT FROM AUTHOR]

Influenza viruses cause respiratory diseases in humans and animals with high morbidity and mortality rates. Conventional anti-influenza drugs are reported to exert side effects and newly emerging viral strains tend to develop resistance to these commonly used agents. Fritillaria thunbergii (FT) is traditionally used as an expectorant for controlling airway inflammatory disorders. Here, we evaluated the therapeutic effects of FT extracts against influenza virus type A (H1N1) infection in vitro, in ovo , and in vivo. In the post-treatment assay, FT extracts showed high CC50 (7,500 µg/ml), indicating low toxicity, and exerted moderate antiviral effects compared to oseltamivir (SI 50.6 vs. 222) in vitro. Antiviral activity tests in ovo revealed strong inhibitory effects of both FT extract and oseltamivir against H1N1 replication in embryonated eggs. Notably, at a treatment concentration of 150 mg/kg, only half the group administered oseltamivir survived whereas the FT group showed 100% survival, clearly demonstrating the low toxicity of FT extracts. Consistent with these findings, FT-administered mice showed a higher survival rate with lower body weight reduction relative to the oseltamivir group upon treatment 24 h after viral infection. Our collective results suggest that FT extracts exert antiviral effects against influenza H1N1 virus without inducing toxicity in vitro, in ovo or in vivo, thereby supporting the potential utility of FT extract as a novel candidate therapeutic drug or supplement against influenza.

Fritillaria thunbergii is a commonly used traditional Chinese medicine, which has the effects of clearing heat and resolving stagnation, eliminating phlegm and relieving cough. At present, it is mostly produced by cultivation, and the cultivation process requires application of base fertilizer, winter fertilizer, seedling fertilizer and late top dressing. Now farmyard manure or organic fertilizer can be used to replace the base fertilizer and winter fertilizer, but the research on the replacement of organic fertilizer has not been completed for the late top dressing. Potassium fulvate is a kind of fulvate fertilizer, which can not only regulate the growth of crops but also supplement potassium necessary for the growth of crops. In this paper, using F. thunbergii as a model plant with mature cultivation techniques, the effect of potassium fulvate on the quality and yield of rhizome traditional Chinese medicine F. thunbergii was systematically studied for the first time. HPLC-ELSD was used to determine the contents of peimine A and peimine B, hot dip method was used to determine the content of alcohol extract, and the SPAD-502 Plus chlorophyll meter was used to detect SPAD value. The results showed that applying 1.5 to 2.25 kg·hm~(-2) of potassium fulvic acid per hectare could effectively improve the yield of F. thunbergii and there was significantly difference between potassium phosphate monobasic and potassium fulvic acid in terms of quality. After the application of range 1.5 to 2.25 kg·hm~(-2) of potassium fulvic acid per hectare, the content of alcohol soluble extract of F. thunbergii was ranged 21.61% to 22.27%, the total amount of peimine A and peimine B were ranged 0.09% to 0.10%. Applying 1.5 to 2.25 kg·hm~(-2) of potassium fulvic acid per hectare could replace the conventional pure chemical fertilizer potassium phosphate monobasic, which could be used as top dressing fertilizer for the cultivation of F. thunbergii.