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italic>Tussilago farfara L. is a perennial herb of Tussilago genus in the Compositae family. Its dried buds and leaves have good biological activities and have a long history of medicinal use in China and Europe. In this paper, we investigated the whole chloroplast genome characteristics, sequence duplication, structural variation and phylogeny of the Tussilago farfara L. After sequencing the Tussilago farfara L. chloroplast genome using Illumination technology, the complete Tussilago farfara L. chloroplast genome was further obtained by assembly and annotation, followed by a series of inverted repeat-large single copy/small single copy region contraction and expansion analysis, genome sequence variation, etc. The sequences of 13 homologous plants downloaded from NCBI were used to construct a neighbor-joining phylogenetic tree. The results showed that the total GC content of the chloroplast genome was 37.4% and the length was 150 300 bp; 125 genes were annotated, including 82 protein-coding genes, 35 tRNAs and 8 rRNAs; 148 (simple sequence repeats, SSR) loci were detected, and the relative synonymous codon usage showed that 31 codons out of 64 codons had a usage of >1. In the phylogenetic analysis, the chloroplast genomes of the seven species of Asteraceae, including the Yulin Tussilago farfara L., were highly conserved, and the sequence variation of the (large single-copy, LSC) and (small single-copy, SSC) regions was higher than that of the (inverted repeat, IR) region. This is in general agreement with the reported phylogeny of Yulin Tussilago farfara L. In this study, we obtained a high quality chloroplast genome and analyzed its genome characteristics, codon preference, SSR characteristics, SC/IR boundary, sequence variation and phylogeny, which can provide a basis for species identification, genetic diversity analysis and resource development of this medicinal plant.
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Objective:To investigate the allelopathic effects of water extracts from rhizosphere soil of three medicinal plants Rehmannia glutinosa，Pinellia ternata and Isatis indigotica on seed germination and seedling growth of Polygala tenuifolia， screen the stubble varieties suitable for crop rotation with P. tenuifolia， and provide some scientific basis for continuous cropping obstacles of P. tenuifolia. Method:The bioassay method was used to study the effects of rhizosphere soil water extracts from three medicinal plants Rehmannia glutinosa，Pinellia ternata and Isatis indigotica at concentrations of 0.3，0.6，0.9 g·mL-1 on the germination of P. tenuifolia seed and seedling growth. Result:The rhizosphere soil water extracts of Rehmannia glutinosa and Pinellia ternata showed basically low-promotion and high-inhibition concentration effects on the final germination rate，germination potential，and germination index of P. tenuifolia seeds，while the water extract of Isatis indigotica showed significant allelopathic inhibition effect. All three rhizosphere soil water extracts showed significant allelopathic inhibition effects on the growth index of P. tenuifolia seedlings. Among them，the rhizosphere soil water extract of Rehmannia glutinosa showed lower inhibitory effect on the plant height and root length of P. tenuifolia seedlings than the other two water extracts. The photosynthetic pigment content，proline（Pro） content，and soluble sugar content of P. tenuifolia chinensis seedlings were the highest under 0.3 g·mL-1 soil water extract of Rehmannia glutinosa， with relatively higher content of soluble protein， and relatively lower content of hydrogen oxide（H2O2）. Under the treatment of 0.9 g·mL-1 soil water extract of Rehmannia glutinosa，P. tenuifolia seedlings had the highest peroxidase（POD） and superoxide dismutase（SOD） activities，low catalase（CAT） activity，and lowest content of malondialdehyde（MDA）. Conclusion:Based on the comprehensive analysis of the above experimental data and allelopathic effects，the water extract of rhizosphere of Rehmannia glutinosa can promote the germination of P. tenuifolia seeds to a certain extent，and lay the foundation for seedling resistance to biochemical stress. Therefore， Rehmannia glutinosa is more suitable for crop rotation with P. tenuifolia.
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Objective: To establish the HPLC fingerprint of wild and cultivated Notholirion bulbuliferum,and recognize them according to the chemical pattern, in the expectation of providing the basis for the quality control and domestication cultivation of N. bulbuliferum of origins. Method: Twenty samples of wild and cultivated N. bulbuliferum collected from different origins were detected by HPLC, and a common mode of fingerprint was established. The Similarity Evaluation System for Chromatographic Fingerprint of TCM (2012A edition) was used to evaluate the similarity of the samples. The differences among the samples were identified by chemical pattern recognition methods, including principal component analysis (PCA),cluster analysis (HCA) and partial least squares discriminate analysis (PLS-DA). Result: The HPLC fingerprint of N. bulbuliferum was obtained,and 26 common peaks were found in the chromatograph. Similarities of all samples were over 0.9,PCA,and HCA and PLS-DA results demonstrated obvious distinctions between wild and cultivated N. bulbuliferum. Eight constituents,such as pcoumaric acid were identified as biomarkers,representing major differences between the two varieties. Conclusion: The HPLC chromatogram of N. bulbuliferum developed in this paper has strong characteristics and repeatability. After being combined with the pattern recognition mode, it can be used as an effective method for evaluating the quality of N. bulbuliferum and distinguishing wild and cultivated N. bulbuliferum,and provide a reference for the quality control and domestication introduction of N. bulbuliferum.
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Objective:To study the effect of different hormone ratios on the callus induction of roots,stems and leaves of Polygala tenuifolia,and determine and analyze the amount of flavonoids in roots,stems and leaves of P. tenuifolia. Method:With MS as the basic medium and roots,stems and leaves of P. tenuifolia sterile seedings as explants,the effects of 2,4-D,NAA and 6-BA on callus induction and flavonoid accumulation in different parts of roots,stems and leaves of P. tenuifolia were determined by orthogonal test. Result:2,4-D,NAA and 6-BA had significant effects on the callus induction rate of roots,stems and leaves of P. tenuifolia. The optimal callus induction combination of leaves was MS+3.0 mg·L-1 2,4-D+1.0 mg·L-1 NAA+1.5 mg·L-1 6-BA,the optimal callus induction combination of stems was MS+1.0 mg·L-1 2,4-D+3.0 mg·L-1 NAA+1.5 mg·L-1 6-BA,the optimal callus induction combination for roots was MS+1.0 mg·L-1 2,4-D+1.0 mg·L-1 NAA+1.0 mg·L-1 6-BA. And 2,4-D,NAA and 6-BA had significant effects on flavonoid accumulation in the stem callus of P. tenuifolia,and MS+3.0 mg·L-1 2,4-D+1.0 mg·L-1 NAA+0.5 mg·L-1 6-BA was the best flavonoid accumulation combination.NAA,6-BA had significant effects on flavonoid accumulation in the leave callus of P. tenuifolia,while 2,4-D had no significant effect,and MS+3.0 mg·L-1 2,4-D+2.0 mg·L-1 NAA+1.0 mg·L-1 6-BA was the optimal flavonoid accumulation combination,the three hormones had no significant effect on the accumulation of flavonoids in the root callus of P. tenuifolia,and MS+2.0 mg·L-1 2,4-D+1.0 mg·L-1 NAA+1.5 mg·L-1 6-BA was the best flavonoid accumulation combination. Conclusion:Under the conditions,the callus induction rate of roots,stems and leaves of P. tenuifolia is 100%, especially, the callus of P. tenuifolia leaves was the optimal,which is followed by P. tenuifolia stems and P. tenuifolia roots. Under the conditions,the amount of flavonoids in roots,stems and leaves of P. tenuifolia reach 21.31,24.56,23.61 mg·g-1,respectively.
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Objective:To study the changes in total phenolic and flavonoid content and antioxidant enzyme activity of Polygala tenuifolia callus in MS medium with different concentrations of H2O2,in order to explore the physiological mechanism of Polygala tenuifolia callus in adapting to H2O2 environmental stress at the cellular level. Method:Five gradients of 0,5,10,15,20 mmol·L-1 were set for H2O2 concentration and added to MS medium, with P. tenuifolia callus as the experimental material. Total phenols,total flavonoids and antioxidant enzyme activities of callus were determined after 5,10,15,20,25 d of culture,respectively. Result:The contents of total phenols and flavonoids were the highest when the concentration of H2O2 was 5 mmol·L-1 for 15 d. The SOD activity was the highest when the callus was cultured for 5 d and the exogenous H2O2 concentration was 5 mmol·L-1. POD activity was the highest at 25 d and 5 mmol·L-1 concentration of exogenous H2O2.CAT activity was the highest at 25 d and 15 mmol·L-1 concentration of exogenous H2O2. Conclusion:P. tenuifolia callus has the ability to adapt to the environmental stress of H2O2 at a certain concentration. When it is subjected to the environmental stress of H2O2,P. tenuifolia callus can alleviate the damage by regulating its secondary metabolites and protecting enzyme system. It can significantly promote the content of total phenols and flavonoids in secondary metabolites at 5-10 mmol·L-1. SOD activity was significantly increased at 5 d and the concentration of exogenous H2O2 of 5 mmol·L-1. POD activity was significantly increased at 25 d and the concentration of exogenous H2O2 of 5 mmol·L-1. CAT activity was significantly increased at 25 d and concentration of exogenous H2O2 of 15 mmol·L-1.
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Shaanxi has a long history and profound cultural heritage. It has been referred to as “Qin” for a long time in history. Shaanxi is rich in medicinal plant resources, various main Chinese medicines such as Fraxini cortex, Gentianae Macrophyllae Radix, Ten “Qin medicine” (Astragali Radix in Zizhou County, Bupleuri Radix in Baoji, Corydalis Rhizoma in Yang County, Salviae Miltiorrhizae Radix et Rhizoma in Shangluo, Aconiti Lateralis Radix Praeparata in Hanzhong; Eucommiae Cortex in lueyang County; Gastrodiae Rhizoma in Ningshan County; Polyporus in Ningshan County; Scutellariae Radix in Chengcheng County; Corni Fructus in Foping County; Polygonati Rhizoma in Lueyang County) and “Taibaiqi medicine” are typically species of “Qin medicine”. In history, “Qin medicine” represents genuine medicinal materials produced from Shaanxi Province and the surrounding region. In recently, the industry of “Qin medicine” becomes one of the most valuable resources in Shaanxi, which also contributes to the integral part of Chinese medical and health services. Here, the research progress on germplasm resources, cultivation techniques, plantation base construction, variety breeding, bioactive compounds, and quality control of “Qin medicine” were integrated and reviewed, and its future development of “Qin medicine” was also prospected.
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Objective To learn the prevalence, distribution and risk factors of mild cognitive impairment (MCI) among elderly in Zhoushan City, and to provide the reference for MCI control. Methods A total of 1801 elderly people aged 60 to 79 years old from six districts of Zhoushan City were sampled by method of stratified random sampling. After self-evaluated with Ascertain Dementia 8 (AD8) and screened with Screening Scale for Mild Cognitive Impairment (sMCI), the diagnosis by specialists was conducted for that positive to AD8 and sMCI. Results of 1801 respondents, 873 (48.47％) people were male, and the other 928 (51.53％) people were female; 38.65％ of the people selected aged 60-<65; 90.28％ had a primary school education or were illiterate; 78.51％ had legitimate and healthy wives. A total of 122 elderly people were diagnosed with MCI, and the prevalence of MCI was 6.77％. Multivariate logistic regression analysis showed that male elderly people (OR=0.53, 95％CI:0.28-1.00) were less likely to develop MCI compared to the female, and the illiterate (OR=2.09, 95％CI: 1.16-3.77) were more likely to develop MCI compared to the educated . Conclusion The prevalence of MCI among the elderly in Zhoushan was 6.77％; the female and the illiterate were more likely to develop MCI.
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Rubia cordifolia L. has been used as a traditional Chinese medicine for several centuries. In recent years, the resources of wild Rubia cordifolia have been declined sharply due to increased utilization and rising price. Therefore, it is of great urgency to evaluate and protect resources of wild plant of Rubia cordifolia. In our study, sixty-four individuals that represent eight wild populations of Rubia cordifolia L. were analyzed by Start Codon Targeted Polymorphism (SCoT) molecular markers. Genetic distance was calculated by POPGENE 3.2 software, cluster analysis was generated by NTSYS 2.10 software based on UPGMA method and Mantel Test was used to analysis the relationship between the genetic distances and geographical distance among the wild populations. The results showed a high genetic diversity of wild populations of Rubia cordifolia L. in Shaanxi province. A total of 182 bands were produced by 14 primers, among which 163 bands were polymorphic bands, and the percentage of polymorphic bands was 89.56%. The average value of Nei's genetic diversity index (H) was 0.293 6, Shannon's information index (I) was 0.444 6, genetic differentiation coefficient (Gst) was 0.555 3, and the gene flow (Nm) was 0.440 8, the wild populations were ranked by genetic diversity:AK > YL > SL > BJ > TC > YA > WN > XY. Mantel Test analysis demonstrated that the significant correlation was found between the genetic distances and geographical distances (r=0.776 4, PRubia cordifolia L. germplasms.
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This study was designed to investigate the molecular mechanism and potential active constituents of Polygala Radix in the treatment of Alzheimer's disease with multiple data bases combined with literature mining to build Polygala Radix chemical composition database. A novel analysis tool Pharmmapper was used to obtain the main active ingredient and potential target of Polygala Radix. By extensive data profiling, the Polygala Radix was found to contain 111 chemical constituents. Among them, a total of 10 active molecules included 3 xanthone, 1 saponins, 3 oligosaccharide esters, and 3 other classes were related to 13 Alzheimer's disease-related targets. Two of the core targets were beta-secretase 1 and glycogen synthase kinase-3 beta. Use the GO analysis and KEGG to explore the molecular mechanism of Polygala Radix in treatment of Alzheimer's disease, which has 3 signaling pathways, and the most important signaling pathway is the cell death signaling pathway. The active constituents of Polygala Radix could control the formation of Aβ and the apoptosis of cells through the interaction with multiple targets, and control the treatment of Alzheimer's disease.
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This work was launched to explore the effect of habitat and growth year on the secondary metabolites contents of cultivated Polygala tenuifolia. The samples of cultivated P. tenuifolia were analyzed by ultra-high performance liquid chromatography(UPLC)-quadrupole time-of-flight mass spectrometry(Q-TOF MS), and the obtained data were analyzed using multiple statistical analysis and cluster analysis. The results showed that compared with growth year, habitat is a main influencing factor which affected the secondary metabolites contents of P. tenuifolia. The contents of sucrose esters and oligosacchride multi-esters are greatly dependent on the habitat (the sample-AG with high levels of components of tenuifoliside B and tenuifoliside C, and the sample-FY with high levels of 3,6'-disinapoyl sucrose, tenuifoliose S, tenuifoliose L, and tenuifoliose V). There is no obvious effect of habitat and growth year on xanthone. The contents of triterpene saponins are greatly dependent on the growth year, and the content of parts of triterpene saponins increased as time goes on.The result indicated that the effect of habitat and growth year on different types of secondary metabolites is not completely equivalent. This study will contribute to the breeding of P. tenuifolia and amendment of current commodity criteria.
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Mitogen-activated protein kinases (MAPKs) are important signaling transduction components well conserved in eukaryotes and play essential roles in various physiological, developmental and hormonal responses in plant. In the present study, a MAPK gene, designated as DoMPK4 (GenBank accession No. JX297597), is identified from a rare endangered medicinal orchid species D. officinale using the reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) methods. The full length cDNA of DoMPK4 is 1 518 bp in length and encoded a 369 aa protein with a molecular weight of 42.42 kD and an isoelectric point of 5.55. DoMPK4 protein contained a serine/threonine protein kinase active site (158-170), a MAP kinase site (71-174), and eight conserved motifs. DoMPK4 had a transmembrane (214-232) but no signal peptide. Multiple sequence alignment showed that DoMPK4 shared high identities (74.9%-80.6%) with MAPK proteins from various plants. Phylogenetic analysis demonstrated that DoMPK4 belonged to group A of the MAPK evolutionary tree, and is closely related to monocots. Real time quantitative PCR (qPCR) analysis revealed that DoMPK4 is differentially expressed among the five organs including leaf, stem, root, seed, and protocorm-like body (PLB). The transcription level of DoMPK4 is the highest in the PLBs with 17.65 fold, followed by seeds, roots, and stems with 5.84, 2.28, and 1.64 fold, respectively. The progressive enhancement of DoMPK4 transcripts in the developing PLBs compared to that in the germinating seeds, suggests a role of DoMPK4 during the development of embryogenic PLBs formation in D. officinale.