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Analyze the changes in gene expression profiles during the process of Panax ginseng seed dormancy release, and screen for differential genes, providing a basis for analyzing the mechanism of P. ginseng seed dormancy release. Comparative transcriptome analysis was conducted by using RNA-Seq sequencing technology in P. ginseng seeds stored at different low temperature. A total of 80.97 Gb of Raw reads and 80.19 Gb of Clean reads were obtained from the transcriptome. Principal component analysis and correlation analysis showed that there were significant differences in gene expression patterns at different developmental stages. Upset results showed that 46 248 unigenes were co-expressed in four stages, and 414, 445, 400 and 389 unigenes were specifically expressed in 0, 8,14 and 28 days, respectively. Gene Ontology functional annotation showed that the differentially expressed genes were mainly involved in nsaturated fatty acid biosynthetic process, nuclear body and oxidoreductase activity. Encyclopedia of Genes and Genomes metabolic pathway showed that differentially expressed genes were mainly involved in peroxisome, mitogen-activated protein kinase signaling pathway-plant, plant hormone signal transduction, ribosome, biosynthesis of unsaturated fatty acid, circadian rhythm-plant and other metabolic pathways. In the process of P. ginseng seed dormancy release, multiple biological processes, such as unsaturated fatty acid biosynthesis and plant hormone signal transduction, are required to coordinate regulation, which constitutes a complex dormancy release regulation network. Transcriptome analysis and differential gene screening of P. ginseng seeds at different sand storage time laid a foundation for the analysis of P. ginseng seed dormancy release mechanism and molecular breeding.
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ObjectiveTo develop safe and effective microbial agents against Panax ginseng root rot. MethodP. notoginseng endophytes were screened in plate confrontation tests， followed by morphological and molecular biological identification of antagonistic strains， optimization of strain fermentation conditions in a single factor test， and determination of optimal carriers and auxiliary agents of the microbial agent and their ratio using response surface methodology for formulating the production process. The prevention and control effects of the microbial agent were verified in the confrontation and pot culture experiments. ResultThe plate confrontation test yielded a strain named Fusarium pseudoanthophilum with significant resistance to root rot， and its antibacterial rate was 53.33%. According to the single factor test， the fermentation conditions of F. pseudoanthophilum were determined to be fermentation time 60 h， fermentation temperature 26 ℃， speed 120 r·min-1， and pH 6.5. The response surface optimization results showed that the number of viable bacteria reached the maximum （5.23×109 cfu·g-1） when the peat was 60.00 g， sodium carboxymethylcellulose 3.50 g， and sodium alginate 4.76 g. The influences of carriers and auxiliary agents on the number of viable bacteria were sorted by degree in a descending order as follows： peat>sodium carboxymethylcellulose >sodium alginate. The confrontation test results showed that when the microbial agent concentration was greater than 1.00 g·L-1， it had a significant inhibitory effect on the root rot pathogen F. oxysporum and the inhibitory rate was more than 42.3%. As demonstrated by the pot culture experiment， the inoculation of biocontrol agent for 28 d significantly reduced the incidence （66.99%） of root rot in P. ginseng seedlings and disease index （61.69%） and increased their leaf length （33.04%） and fresh weight （34.48%）. ConclusionF. pseudoanthophilum inoculant is efficient in preventing and controlling the root rot， making it worthy of further development and utilization.
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The development of quality Chinese medicine is an important way to improve the quality of Chinese medicine, and ensure the safety and effectiveness of Chinese medicine. This article systematically elaborates the definition, classification, standard and mana-gement certification strategy of quality Chinese medicine. We present the quality Chinese medicine which is higher quality than that of eligible Chinese medicine based on quality control standards. Quality Chinese medicine is strictly in accordance with management procedures, likely GAP and GMP et al, during the productive process, which quality indicators is higher than that of the current relevant national quality standards, such as Chinese Pharmacopoeia(ChP) et al; its limited indicators such as exogenous pollutants and endogenous toxic substances are lower than that of the current relevant national quality standards, likely ChP et al; meanwhile these Chinese herbal medicine, medicinal pieces, patent medicines, and health products and foods with Chinese medicine raw materials are been certificated by quality Chinese medicine. At the same time, this article systematically expounds the five major management systems of quality Chinese medicine, including technical training management for practitioners, productive process management, standard mana-gement, quality inspection and certification management, and product traceability management. And we put forward strategies to improve the supervision and management system, and promote the standardization and development of quality Chinese medicine by improving the technical management system of quality Chinese medicine, strengthening the quality management system and six sigma(6σ) management in the company. These strategies will provide a reliable basis and effective way to improve the quality of Chinese medicine.
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Drugs, Chinese Herbal , Food , Medicine, Chinese Traditional , Quality Control , Reference StandardsABSTRACT
The excessive pesticide residues and heavy metals in traditional Chinese medicine seriously endanger human health and the sustainable development of Chinese medicine industry. In order to improve the quality of traditional Chinese medicine and establish a general standard for maximum residue limits(MRL) of pesticides in pollution-free traditional Chinese medicine and decoction pieces, and to ensure the safety of clinical medication from its origin, MRLs were calculated based on the formula(MRL=A×W/100M) from Chinese Pharmacopeia, comparing it with the current Chinese and international standards as well as literature review, the RAND/UCLA appropriateness method(RAM) was applied to determine the categories and MRLs of pesticides in pollution-free traditional Chinese medicine and decoction pieces. Two questionnaires were drafted for expert panel and appropriateness analysis was carried out with the 9-point Likert scale to determine the general standard for MRLs of pollution-free traditional Chinese medicine and decoction pieces. The results showed that a total of nine experts from different fields scored the necessity of standard-setting and 206 pesticide residue limits respectively. The appropriateness scores of 206 pesticides were greater than 7, and appropriateness rate was 100%, which signifies that the expert panel has reached consensus. In summary, based on the RAM, the general standard for maximum residue limits of pesticides in pollution-free Chinese medicines and decoction pieces has reached an expert consensus. Comparing with the MRLs of medicinal plants and plant-sourced food from CAC, Europe Union, the United States, South Korea, Japan, Australia, New Zealand and Canada, 206 MRLs from this general standard share 88.8% in common, 4.4% of which is higher and 6.8% lower than those international standards. This has provided a basis for standardizing the use of pesticides in pollution-free traditional Chinese medicine.
Subject(s)
Drugs, Chinese Herbal , Reference Standards , Medicine, Chinese Traditional , Pesticide Residues , PesticidesABSTRACT
Aimed to solve the issues of pesticide residue, heavy metal contents and harmful elements in the productive process of Chinese herbal medicines, the research team built the technical regulations for production of pollution-free Chinese herbal medicines. This regulation included the environment of production area, the process of production, quality of products etc., which were the key steps controlled the quality of Chinese herbal medicines. The environment of production area was selected according to the ecological factors which were stipulated by Ecological Suitability Regionalization of Chinese herbal medicines (second edition). The quality of air should be attain the one or two levels of GB/T3095-2012 standard values. The cultivation soils should reach to the one or two levels of GB15618 and NY/T391 standard values. The quality of irrigation water should accord with the stipulation of GB5084-2005. Aimed to the production of Chinese herbal medicines, disease-resistant and superior varieties which were suitable to the local stations should be selected, and the breeding of superior seeds and seedlings should be strengthened. Additionally, rational fertilizer application of pollution-free Chinese herbal medicines should be conformed to the principles, requirements, and the kinds of fertilizers allowed or limited for use, which were stipulated by the standard of DB13/T454. Furthermore, the plant protection policy of priority to prevention and synthetical prevention should be followed; improving ecological environment and strengthening cultivation management should be served as the basics. Agricultural measures, and biological and physical control strategies should be preferred to use; and high toxicity, residue pesticide and its mixture should be inhibited; the use of chemical pesticides should be minimized and then to decrease contamination and residue. Additionally, the quality of products should be reached to the standard of pollution-free Chinese herbal medicines; high toxicity and detection rate of pesticide residues and the contents of heavy metal and harmful elements (e.g. plumbum, cadmium, mercury, arsenic and cuprum) should accord with the common criteria of pollution-free Chinese herbal medicines. Application of technical regulations for production of pollution-free Chinese herbal medicines guarantees significantly social, economic and ecological benefits.
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DNA marked-assisted selection of medicinal plants accelerated the breeding and promotion of new cultivars, and guaranteed the healthy development of Chinese medicinal materials industry. The first disease-resistant cultivar of notoginseng, namely "Miaoxiang Kangqi 1", served as the object of study. We evaluated the Kangqi's resistance of seeds, seedlings and root against the pathological bacteria (Fusarum oxysporum) of root rot. Compared to the traditional cultivars, the disease index of notoginseng seeds declined by 52.0% after inoculation for seven days; the death rate of seedlings and disease index of root respectively decreased by 72.1% and 62.4% after inoculation for 25 days. Additionally, the growth inhibition ratio of notoginseng seeds and seedlings declined after inoculation. The seeds, seedlings and roots of "Miaoxiang Kangqi 1" showed significantly resistant to root rot. The evaluation of disease-resistance of Kangqi provided the basis for the popularization of new cultivar and guaranteed the favoring conduct of notoginseng pollution-free cultivation.
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DNA marker-assisted selection of medicinal plants is based on the DNA polymorphism, selects the DNA sequences related to the phenotypes such as high yields, superior quality, stress-resistance and so on according to the technologies of molecular hybridization, polymerase chain reaction and high-throughput sequencing, and assists the breeding of new cultivars. This study bred the first disease-resistant cultivar of notoginseng "Miaoxiang Kangqi 1" using the technology of DNA marker-assisted selection of medicinal plants and systematic breeding. The disease-resistant cultivar of notoginseng contained 12 special SNPs based on the analysis of Restriction-site Associated DNA Sequencing (RAD-Seq). Among the SNP (record_519688) was related to the root rot-resistant characteristics, which indicated this SNP could serve as genetic markers of disease-resistant cultivars and assist the systematic breeding. Compared to the conventional cultivated cultivars, the incidence rate of root-rot and rust-rot in notoginseng seedlings decreased by 83.6% and 71.8%, respectively. The incidence rate of root-rot respectively declined by 43.6% and 62.9% in notoginseng cultivation for 2 and 3 years compared with those of the conventional cultivated cultivars. Additionally, the potential disease-resistant groups were screened based on the relative SNP, and this model enlarged the target groups and advanced the breeding efficiency. DNA marker-assisted selection of medicinal plants accelerated the breeding and promotion of new cultivars, and guaranteed the healthy development of Chinese medicinal materials industry.
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Objective: To research the influence of different washing methods before drying on the contents of active components in Salvia Miltiorrhizae Radix et Rhizoma, and provide the basis for standard original processing methods of Salvia miltiorrhiza. Methods: The fresh herbs of S. miltiorrhiza were processed with different methods including no washing, water flushing, immersion cleaning, and scrubbing in water. HPLC was applied to determine the contents of active components, with Diamonsil C18 column (250 mm×4.6 mm, 5 μm), acetonitrile-0.026% phosphoric acid for gradient elution, flow rate of 1 mL/min, and detection wave length of 286 and 270 nm. Results: The contents of rosmarinic acid, salvianolic B, crypotanshinone, tanshinone I, and tanshinone IIA were all significantly decreased in S. miltiorrhiza processed with different washing methods. The contents of active components were reduced with the increasing of washing intensity, of which the lowest contents were found in scrubbing group. Conclusion: Washing with water can make loss of active components, and flushing may be a more suitable method. In order to decrease the content loss of active components, contact time with water must be controlled as short as possible and violent friction among medicinal materials and artificial rubbing must be avoided.