[Effects of organic fertilizer from traditional Chinese medicine residues on growth and soil microbial community of Salvia miltiorrhiza by metagenomic technique].

Soil microbiome is a key evaluation index of soil health. Previous studies have shown that organic fertilizer from traditional Chinese medicine(TCM)residues can improve the yield and quality of cultivated traditional Chinese medicinal materials. However, there are few reports on the effects of organic fertilizer from TCM residues on soil microbiome. Therefore, on the basis of evaluating the effects of organic fertilizer from TCM residues on the yield and quality of cultivated Salvia miltiorrhiza, the metagenomic sequencing technique was used to study the effects of organic fertilizer from TCM residues on rhizosphere microbiome community and function of cultivated S. miltiorrhiza. The results showed that:(1) the application of organic fertilizer from TCM residues promoted the growth of S. miltiorrhiza and the accumulation of active components, and the above-ground and underground dry weight and fresh weight of S. miltiorrhiza increased by 371.4%, 288.3%, 313.4%, and 151.9%. The increases of rosmarinic acid and salvianolic acid B were 887.0% and 183.0%.(2)The application of organic fertilizer from TCM residues significantly changed the rhizosphere bacterial and fungal community structures, and the microbial community composition was significantly different.(3)The relative abundance of soil-beneficial bacteria, such as Nitrosospira multiformis, Bacillus subtilis, Lysobacter enzymogenes, and Trichoderma was significantly increased by the application of organic fertilizer from TCM residues.(4)KEGG function prediction analysis showed that metabolism-related microorganisms were more easily enriched in the soil environment after organic fertilizer application. The abundance of functional genes related to nitrification and denitrification could also be increased after the application of organic fertilizer from TCM residues. The results of this study provide guidance for the future application of organic fertilizer from TCM residues in the cultivation of traditio-nal Chinese medicinal materials and enrich the content of green cultivation technology of traditional Chinese medicinal materials.

[Systematic identification of chemical forms of key terpene synthase in Cinnamomum camphora].

Cinnamomum camphora is an important economic tree species in China. According to the type and content of main components in the volatile oil of leaf, C. camphora were divided into five chemotypes, including borneol-type, camphor-type, linalool-type, cineole-type, and nerolidol-type. Terpene synthase(TPS) is the key enzyme for the formation of these compounds. Although several key enzyme genes have been identified, the biosynthetic pathway of(+)-borneol, which has the most economic value, has not been reported. In this study, nine terpenoid synthase genes CcTPS1-CcTPS9 were cloned through transcriptome analysis of four chemical-type leaves. After the recombinant protein was induced by Escherichia coli, geranyl pyrophosphate(GPP) and farnesyl pyrophosphate(FPP) were used as substrates for enzymatic reaction, respectively. Both CcTPS1 and CcTPS9 could catalyze GPP to produce bornyl pyrophosphate, which could be hydrolyzed by phosphohydrolase to obtain(+)-borneol, and the product of(+)-borneol accounted for 0.4% and 89.3%, respectively. Both CcTPS3 and CcTPS6 could catalyze GPP to generate a single product linalool, and CcTPS6 could also react with FPP to generate nerolidol. CcTPS8 reacted with GPP to produce 1,8-cineol(30.71%). Nine terpene synthases produced 9 monoterpene and 6 sesquiterpenes. The study has identified the key enzyme genes responsible for borneol biosynthesis in C. camphora for the first time, laying a foundation for further elucidating the molecular mechanism of chemical type formation and cultivating new varieties of borneol with high yield by using bioengineering technology.

Traditional Chinese medicine residues promote the growth and quality of Salvia miltiorrhiza Bunge by improving soil health under continuous monoculture.

Continuous monoculture of crops has resulted in reduced yields and quality, as well as soil deterioration. Although traditional Chinese medicine residues (TCMRs) are known to promote plant growth and soil health, few studies have investigated their effectiveness in continuous monoculture soils. Here, we studied the impact of chemical fertilizers (CF) and four TCMRs with antibacterial activities on the growth of S. miltiorrhiza (a widely used medicinal plant in China), accumulation of active ingredients in plants, and soil health under continuous monoculture conditions. Compared with no fertilizer (CK) and CF, fermented Sophora flavescens radix residue (SFRf) and fermented and unfermented Moutan cortex residue (MCRf and MCRu, respectively) resulted in a reduction of the disease index of root rot, while CF did not. The CF and four TCMR treatments increased the accumulation of nitrogen (N) (42.8-124.6% and 17.0-101.7%), phosphorous (P) (19.8-74.7% and 8.3-27.4%), and potassium (K) (104.1-212.0% and 9.3-51.8%) in shoots and roots compared to CK. The differences in nutrient accumulation between the CF and TCMR treatments were statistically insignificant, excepted for the N accumulation in the roots. All fertilization treatments increased plant biomass compared to CK, with increases of 25.57-89.86% and 2.62-35.28% in shoots and roots, respectively. The SFRf treatment exhibited the most significant enhancement in both shoot and root biomass. CF significantly reduced the accumulation of seven active ingredients in roots by 23.90-78.95% compared to CK, whereas each TCMR increased accumulation of certain active ingredients. The TCMR treatments effectively improved the health of deteriorated soil by enhancing soil physicochemical properties, restoring the balance of the microbial community, recruiting beneficial bacteria, and reducing the relative abundance of the pathogen Fusarium. The SFRf treatment exhibited superior performance in improving soil health than other treatments. Overall, the TCMRs outperformed CF in restoring soil health and promoting the yield and quality of S. miltiorrhiza. These findings offer guidance for improving the health of continuous cropping soil and recycling TCMRs.

[Effect of Rhizophagus intraradices on growth of Salvia miltiorrhiza].

The study aimed to explore the effects of inoculation of Rhizophagus intraradices on the biomass, effective component content, and endogenous hormone content of Salvia miltiorrhiza through pot experiments. The number of leaves, plant height, dry weight of aboveground and underground parts, branch number, root number, root length, root diameter, and other biomass were mea-sured by weighing and counting methods. The content of salvianolic acid B, caffeic acid, rosmarinic acid, tanshinone Ⅰ, tanshinone Ⅱ_A, cryptotanshinone, and other effective components was determined by ultra-high performance liquid chromatography. The content of ABA and GA_3 was determined by triple quadrupole mass spectrometry. The correlations between biomass and effective components and between effective components and plant hormones ABA and GA_3 were analyzed. The results showed that R. intraradices significan-tly increased the aboveground dry weight, leaf number, and root number of S. miltiorrhiza by 0.24-0.65 times, respectively. The content of salvianolic acid B and rosmarinic acid in the aboveground part and the content of salvianolic acid B, caffeic acid, rosmarinic acid, tanshinone Ⅰ, and tanshinone Ⅱ_A in the underground part were significantly increased by 0.44-1.78 times, respectively. R. intraradices infection significantly increased the GA_3/ABA values of aboveground and underground parts by 3.82 and 76.47 times, respectively. The correlation analysis showed that caffeic acid, the effective component of the aboveground part, was significantly positively correlated with plant height, tanshinone Ⅱ_A, the effective component of the underground part, was significantly positively correlated with biomass root number, cryptotanshinone, and dry weight, while rosmarinic acid was significantly negatively correlated with dry weight. There were significant positive correlations between cryptotanshinone and ABA, tanshinone Ⅱ_A and ABA and GA_3, and caffeic acid and GA_3. In conclusion, R. intraradices can promote the accumulation of biomass and secondary metabolites of S. miltiorrhiza and regulate the balance between plant hormones ABA and GA_3, thereby promoting the growth of S. miltiorrhiza.

[Effects of temperature and humidity on infection of Fusarium oxysporum in seedlings of Salvia miltiorrhiza].

Wilt disease is a major disease of cultivated Salvia miltiorrhiza, which is caused by Fusarium oxysporum. Since the infection process of F. oxysporum in plants is affected by environment factors, this study was conducted to reveal the relationship between disease severity and concentration of the pathogen in plants in the infection process of F. oxysporum in seedlings of S. miltiorrhiza by pot experiments and to reveal the effects of temperature and humidity on the infection process. The results showed that, after inoculation of S. miltiorrhiza seedlings with F. oxysporum, the pathogen in different parts was detected at different time, and it was first detected in substrates. With the continuous propagation of the pathogen(4-5 d), it gradually infected the roots and stems of the seedlings, and the plants had yellowing leaves and withering. The number of the pathogen reached the maximum in each part after 7-8 d, and then gradually decreased in the later stage of the disease. The concentration of the pathogen in substrates, roots and stems of S. miltiorrhiza showed a trend of decreasing after increasing with the aggravation of the disease and reached the maximum in the samples of moderate morbidity, while the concentration in the samples of severe morbidity decreased. In addition, the infection of F. oxysporum in seedlings of S. miltiorrhiza was affected by temperature and humidity. The suitable temperature was 25-30 ℃ and the suitable humidity was 80%-90%. This study could provide guidance for the experiments on pathogenicity of F. oxysporum, screening of biocontrol bacteria and controlling of wilt.

[Chemical and nutrient differences between medicinal material, residues, and residue compost of Moutan Cortex].

Moutan Cortex(MC) residues produced after the extraction of MC can be re-extracted for active components and used to produce organic fertilizer and animal feed. However, they are currently disposed as domestic waste, which pollutes the environment. This study analyzed the chemical composition of the medicinal material, residues, and residue compost of MC by UPLC-UV-Q-TOF-MS. Furthermore, the nutrient composition of MC residues and the residue compost was analyzed. The results showed that:(1)MC residues had lower content of chemicals than the medicinal material, and content of paeonol, gallic acid, and galloylglucose in MC residues were about 1/3 of that in the medicinal material. The content of chemicals were further reduced after residue composting, and the quantitative compounds were all below the limits of detection.(2)Compared with MC residues, the residue compost showed the total nitrogen, total phosphorus, total potassium, and organic matter content increasing by 122.67%, 31.32%, 120.39%, and 32.06%, respectively. Therefore, we concluded that the MC residues can be used to re-extract active compounds such as paeonol, gallic acid, and galloylglucose. The MC residue compost is a high-quality organic fertilizer containing minimal content of chemicals and can be widely used in the cultivation of Chinese medicinal herbs.

[Pathogenicity and induced systemic resistance of Fusarium oxysporum and Verticillium dahlia to Salvia miltiorrhiza].

Salvia miltiorrhiza is a commonly used bulk medicinal material in China. Due to the increasing demand in recent years, the planting area is expanding. In the artificial cultivation of S. miltiorrhiza, continuous cropping obstacles are prominent, which has seriously restrained the growth of S. miltiorrhiza, resulted in serious root diseases, and affected the yield and quality of medicinal materials. The pathogen infection can induce plant resistance. Previously, this research group isolated Fusarium oxysporum and Verticillium dahlia from the roots of diseased S. miltiorrhiza. In this study, 7 days after inoculation of S. miltiorrhiza with F. oxysporum(Foc group) and V. dahlia(Vd group), the incidence rates in S. miltiorrhiza were 48% and 26%, respectively. Both the two pathogens significantly reduced the aboveground biomass of S. miltiorrhiza. Five days after inoculation, the activities of defensive enzymes, such as peroxidase(POD), phenylalanine ammonia-lyase(PAL), superoxide dismutase(SOD), and polyphenol oxidase(PPO) reached the peak. The enzyme activity of the Foc group was significantly higher than that of the Vd group. Three days after inoculation, the expression of defense genes SmPDF2.1 and SmPR10 peaked and then decreased. The results showed that F. oxysporum and V. dahlia showed pathogenicity to S. miltiorrhiza and could strongly induce systemic resistance. In terms of the above indexes, F. oxysporum was superior to V. dahlia.

[Effects of interaction between ubiquitin/26S proteasome inhibitor MG132 and mycorrhiza on growth and effective components of Salvia miltiorrhiza].

Ubiquitin/26 S proteasome system(UPS) is one of the main ways to regulate the degradation of proteins in plants, and plays an important role in physiological processes such as secondary metabolism and plant hormone signal transduction. As indicated recently, UPS is involved in plant-microbe interactions, and presumably regulates arbuscular mycorrhizal symbiosis to affect its effects. This study investigated the effects of interaction between Cbz-leu-leu-leucinal(MG132) and the mycorrhiza on the growth and effective components of Salvia miltiorrhiza by inoculation with Glomus intraradices and spraying MG132 solution. The results showed that the inoculation with G. intraradices could promote the growth of S. miltiorrhiza, increase the accumulation of effective components in the aerial and underground parts, and decrease the relative expression level of JMT. Additionally, MG132 could strengthen the growth-promoting effect of G. intraradices. As compared with the control group, the inoculation with G. intraradices could significantly increase aerial and underground fresh weights by 267% and 95%, respectively, under the treatment with MG132 spraying, while under the MG132 spraying-free condition, the increase was 195% and 32%, respectively. Meanwhile, MG132 spraying could enhance the promotion of mycorrhizal fungi on the accumulation of active components of S. miltiorrhiza. On the other hand, regardless of inoculation with G. intraradices or not, MG132 treatment could promote the root division of S. miltiorrhiza, reduce the content of effective components in the aerial parts, and increase the content in the underground part. The inoculation with G. intraradices could alleviate the inhibitory effect of MG132 on the accumulation of effective components in the aerial part of S. miltiorrhiza. The results show that arbuscular mycorrhizal fungi(AMF) can promote the growth of S. miltiorrhiza and the accumulation of effective components, and MG132 treatment can strengthen such promotion effect, which lays a foundation for the application of MG132 in the mycorrhizal cultivation of S. miltiorrhiza in the future.

Salvia miltiorrhiza-asarum ointment combined with Chinese medical massage alleviates symptoms of osteoarthritis in a rat model through the Notch1/ matrix metalloproteinase-13 signaling pathway.

OBJECTIVE: This study investigated the effect of salvia miltiorrhiza-asarum ointment (SMAO) plus Chinese medical massage on knee osteoarthritis in a rat model. METHODS: Hulth's method was used to establish a Sprague-Dawley rat model of knee osteoarthritis (OA). The levels of matrix metalloproteinase-13 (MMP-13), collagen-II, aggrecan, interleukin (IL)-1ß, tumor necrosis factor-α (TNF-α), and IL-6 were measured by enzyme-linked immunosorbent assays. The joint space was assessed by a Perlove X-ray system. Histopathology was examined by hematoxylin-eosin and Safranin O staining. The mRNA and protein expression levels of Notch1, MMP-13, collagen-II, and aggrecan were measured by quantitative reverse transcription-polymerase chain reaction and Western blotting, respectively. RESULTS: SMAO plus Chinese medical massage significantly decreased the levels of MMP-13, IL-1ß, TNF-α, and IL-6, and increased serum collagen-II and aggrecan levels. Pathological injury of the knee joint was improved by SMAO treatment. mRNA and protein expression of Notch1 and MMP-13 was remarkably downregulated, but collagen-II and aggrecan levels were significantly upregulated in cartilage tissues. CONCLUSION: SMAO combined with Chinese medical massage effectively relieves OA symptoms, which may involve inhibiting inflammation through the Notch1/MMP-13 signaling pathway.

[Study on tanshinones regulating root-associated microbiomes of Salvia miltiorrhiza].

The plant root-associated microbiomes include root microbiome and rhizosphere microbiome, which are closely related to plant life activities. Nearly 30% of photosynthesis products of plants are used to synthesize root compounds, there is evidence that root compounds regulate and significantly affect the root microbiome Tanshinones are the main hydrophobic components in Salvia miltiorrhiza. In order to study whether these compounds can regulate the root-associated microbiomes of S. miltiorrhiza, our study first identified a white root S. miltiorrhiza(BG) which contains little tanshinones. Retain of the fifth intron of tanshinones synthesis key enzyme gene SmCPS1 leading to the early termination of the SmCPS1 gene, and a stable white root phenotype. Further, wild type(WT) and BG were planted in greenhouse with nutrient soil(Pindstrup, Denmark) and Shandong soil(collected from the S. miltiorrhiza base in Weifang, Shandong), then high-throughput sequencing was used to analyze the root-associated microbiomes. The results showed that the tanshinones significantly affected the root-associated microbiomes of S. miltiorrhiza, and the impact on root microbiomes was more significant. There are significant differences between WT and BG root microbiomes in species richness, dominant strains and co-occurrence network. Tanshinones have a certain repelling effect on Bacilli which belongs to Gram-positive, while specifically attract some Gram-negative bacteria such as Betaproteobacteria and some specific genus of Alphaproteobacteria. This study determined the important role of tanshinones in regulating the structure of root-associated microbiomes from multiple angles, and shed a light for further improving the quality and yield of S. miltiorrhiza through microenvironment regulation.

Dose-Response Association of Tai Chi and Cognition among Community-Dwelling Older Adults: A Systematic Review and Meta-Analysis.

Previous studies indicated that Tai Chi might be an effective way to improve or prevent cognitive impairments in older populations. However, existing research does not provide clear recommendations about the optimal dose of Tai Chi practice, which is the most effective in improving cognitive function in older adults. The purpose of this systematic review and meta-analysis was to investigate the dose-response relationship between Tai Chi and cognition in community-dwelling older adults. A total of 16 studies with 1121 subjects were included in this study. Meta-regression analyses of Tai Chi duration (Tai Chi session duration, Tai Chi practice duration per week, study duration, and Tai Chi practice duration for the entire study) on the study effect size (ES) were performed to examine the dose-response association of Tai Chi and cognition. The results showed that there was a positive effect of Tai Chi on cognitive function, but there were no statistically significant dose duration effects on cognition. The findings suggest that Tai Chi has beneficial effects on cognitive function, but a longer duration was not associated with larger effects. In order to establish evidence-based clinical interventions using Tai Chi, future research should clearly demonstrate intervention protocol, particularly the style and intensity of Tai Chi.

[Effect of different fungicides on efficiency of mycorrhizal symbiosis on Salvia miltiorrhiza].

Arbuscular mycorrhizal fungi provided is beneficial to Salvia miltiorrhiza for increasing yield, promoting the accumulation of active ingredients, and alleviating S. miltiorrhiza disease etc. However, the application of fungicides will affect the benefit of arbuscular mycorrhizal fungi and there is little research about it. This article study the effect of four different fungicides: carbendazim, polyoxin, methyl mopazine, and mancozeb on mycorrhiza benefit to S. miltiorrhiza by the infection intensity of arbuscular mycorrhizal fungi, the growth of S. miltiorrhiza, and the content of active ingredients. RESULTS:: showed that different fungicides had different effects. The application of mancozeb had the strongest inhibitory effect on the mycorrhizal benefit to S. miltiorrhiza. Mancozeb significantly reduced the mycorrhizal colonization and the beneficial effect of arbuscular mycorrhizal fungi on the growth and the accumulation of active components of S. miltiorrhiza. The application of polyoxin had no significant effect on mycorrhizal colonization. Instead, it had a synergistic effect with the mycorrhizal benefit to promoting the growth and accumulation of rosmarinic acid of S. miltiorrhiza. The inhibitory strengths of four fungicides are: mancozeb>thiophanate methyl, carbendazim>polyoxin. Therefore, we recommend applying biological fungicides polyoxin and avoid applying chemical fungicides mancozeb for disease control during mycorrhizal cultivation of S. miltiorrhiza.

[Cloning and expression analysis of 5-phosphomevalonate kinase gene (CcPMK) in Cinnamomum camphora].

The 5-phosphomevalonate kinase(PMK) is a key enzyme in mevalonate(MVA) pathway which reversibly catalyzes the phosphorylation of mevalonate 5-phosphate(MVAP) to form mevalonate-5-diphosphate(MVAPP) in the presence of ATP and divalent metal ion such as Mg~(2+). In this research, on the basis of the transciptome database of Cinnamomum camphora, the PMK was cloned by cDNA from C. camphora, and was named CcPMK(GenBank number KU886266). The ORF of CcPMK was composed of 1 545 bp, encoding 514 amino acids. The bioinformatics analysis of CcPMK indicated that the molecular weight of the encoded protein was 56.14 kDa, with a theoretically isoelectric point of 7.64, and there was no signal peptide and transmembrane structure in putative protein. By multiple sequence alignment and phylogenetic tree analysis, we found that similarity between CcPMK and PMK amino acid sequence of other plants was as high as 75%. Among the similar sequences, 45% of them belonged to the alpha helix, while 16% belonged to the beta strand. CcPMK obtained 3 PMK protein family motifs and 1 ATP binding site Gly-Leu-Gly-Ser-Ser-Ala-Ala, and its 3 D structure contained a catalytic pocket structure, proving CcPMK as a member of PMK gene family. The result of phylogenetic tree showed that CcPMK was closely related to monocotyledon plants such as Phonenix dactylifera. The results of the Real-time PCR indicated that the expression level of CcPMK in borneol type was higher than that in linalool type, cineol type, iso-nerolidol type and camphor type. CcPMK expressed highest in roots and lowest in branches. Our results revealed that the expression level of CcPMK was different among five chemical types and different plant tissues, and the research provides foundation for further study of the terpenoids biosynthetic pathway in C. camphora.

[Effects of fresh-cut on drying rate and quality of Panax notoginseng].

To study the effects of fresh-cut drying methods on the appearance and internal components of Panax notoginseng, and explore the feasibility of fresh-cut drying methods of P. notoginseng, so as to provide more effective processing methods for the production of P. notoginseng slices and Chinese patent medicines. In this study, we have compared the effects of 6 different drying methods on drying time, drying rate, density, appearance and internal components of P. notoginseng roots. It takes about 453 h to dry by whole-root drying in the sun, with a long constant speed period and a slow drying rate, the time of whole-root drying at 50 ℃ was shortened by 61.6% compared with whole-root drying in the sun, which resulted in the decrease of density and poor appearance of the medicinal material with hollow and crack appeared in the xylem, while the drying time of fresh-cut drying method was reduced by 61.82% to 91.58% and the drying rate increased greatly, due to the relatively slow drying process in the sun or in the shade after fresh-cut, salting-out and whitening appeared on the surface, and the internal components were all decreased to some extent. The drying time of fresh-cut drying at 50 ℃ was 91.58% and 68.83% shorter than that of whole-root drying in the sun and at 50 ℃, respectively. When drying at 50 ℃ after fresh-cut, the appearance and content of internal components of the medicinal materials were better, the appearance was yellowish green, the cut sections were clear with uniform pore distribution, and the content of saponin components was increased by 7.24% compared with that of the whole-root drying at 50 ℃, When drying at 40 ℃, the surface of slices has salting-out and whitening spots, and the loss of dencichine and total sugar was large, but at 60 ℃, this high temperature made the rate of dehydration of slices was extremely fast, which led to severe cracking and fragmentation, and the loss of total sugar and alcohol extract was large. By vacuum freeze drying after fresh-cut, the structure of medicinal materials slices was loose, the density was greatly reduced, and the appearance was different from those recorded in traditional books. The contents of total saponin components and dencichine were increased by 16.51% and 22.54%, respectively, compared with traditional whole-root drying. The fresh-cut process method is feasible in the production of P. notoginseng slices. In production, it is recommended that drying at 50 ℃ after fresh-cut can make the medicinal materials better in appearance and content of internal components, which is convenient for the subsequent processing and industrial feeding extraction. For the purpose of internal contents, it is better to adopt freeze-drying after fresh-cut processing method.

[Effect of soil pH on efficiency of mycorrhizal symbiosis on Salvia miltiorrhiza].

By comparing the effects of soil pH on the efficiency of mycorrhizal symbiosis on Salvia miltiorrhiza， the study is aimed to provide guidance for the use of mycorrhiza in the cultivation of S. miltiorrhiza. In this experiment, the inoculant treated and the non-inoculant treated control were grown in different soil pH. The data was collected after 60 days of cultivation including rate of mycorrhizal infection, biomass, and three chemical constituents with known medicinal action. The results showed that Glomus versiforme was more apt to infect S. miltiorrhiza (F>94.00%; M>69.45%; m>73.66%) and promote the growth of S. miltiorrhiza under pH 5-9 soil. The mycorrhizal contribution to the growth of S. miltiorrhiza was the highest when grown in pH 8 soil. Plants grown with mycorrhiza in pH 8 soil had above-ground biomass more than 2 times and root biomass more than 5 times. The uninoculated plants grew better under acidic and neutral conditions, but the inoculated plants grew better under alkaline (pH 8) conditions. This result showed mycorrhiza can play a role in the adaptability of S. miltiorrhiza to the environment. Inoculation of mycorrhiza significantly increased the accumulation of rosmarinic acid, salvianolic acid B, and dihydrotanshinone by 6.59,5.03 and 2.20-folds. Based on our results alkaline soil (pH 8) is most suitable for the cultivation of S. miltiorrhiza by inoculation with the mycorrhiza G. versiforme.

[Determination of eight steroidal saponins in 15 kinds of genus Paris].

The study was established an HPLC method for simultaneous determination of 8 steroidal saponins (polyphyllins Ⅶ, H, Ⅵ, Ⅱ, Ⅰ, and Ⅴ, dioscin, and gracillin) in Paridis Rhizoma, and made an evaluation by determining steroidal saponins in 15 kinds of genus Paris. The analysis was performed on a Waters Acquity H-ClassTM UPLC ultrafine liquid chromatography system coupled with a PDA detector. The chromatographic separation was achieved through a CAPCELL PAK ADME (4.6 mm× 250 mm, 5 µm) column and the optimal mobile phase consisted of acetonitrile and water. The column was maintained at 21 ℃, and the flow rate was 0.8 mL•min ⁻¹. The UV detection wavelength was 203 nm. The results showed that ① the detected components can be well separated and all with good correlation coefficients. The standard calibration curves were linearly good (R2>0.999 9). The linearity was obtained over 0.041 70-3.812 00 µg. The average recoveries ranged from 95.91% to 103.8%. ② there are significant differences in the content of steroidal saponins from different species. The steroidal saponins are low content or almost none in P. mairei, P. polyphylla var. stenophylla, and P. delavayi have low content or almost did not contain, so these species are not suitable for medicinal use. The contents of steroidal saponins in P. polyphylla var. chinensis are varied from different places. There were high content of steroidal saponins in P. polyphylla var. yunnanensis, P. forrestii, P. daliensis, and P. axialis, even up to 5.0%, which indicated that they had the potential pharmic value of development.

[Climate change impacts on yield of Cordyceps sinensis and research on yield prediction model of C. sinensis].

Cordyceps sinensis is a Chinese unique precious herbal material, its genuine producing areas covering Naqu, Changdu in Qinghai Tibet Plateau, Yushu in Qinghai province and other regions. In recent 10 years, C. sinensis resources is decreasing as a result of the blindly and excessively perennial dug. How to rationally protect, develop and utilize of the valuable resources of C. sinensis has been referred to an important field of research on C. sinensis. The ecological environment and climate change trend of Qinghai Tibet plateau happens prior to other regions, which means that the distribution and evolution of C. sinensis are more obvious and intense than those of the other populations. Based on RS (remote sensing)/GIS(geographic information system) technology, this paper utilized the relationship between the snowline elevation, the average temperature, precipitation and sunshine hours in harvest period (April and may) of C. sinensis and the actual production of C. sinensis to establish a weighted geometric mean model. The model's prediction accuracy can reach 82.16% at least in forecasting C. sinensis year yield in Naqu area in every early June. This study can provide basic datum and information for supporting the C. sinensis industry healthful, sustainable development.

[Advance in studying early bolting of Umbelliferae medicinal plant].

The early bolting has become one of the key issues affecting the Umbelliferae herbs production. The early bolting does not only reduce herbs production but also affect the active ingredients of plants. The concept of early bolting, physiological theory, induce factors of the quality of medicine and, control technology about the early bolting of Umbelliferae are integrative reviewed in the paper. Its occurrence mechanism include: genetic factors, environmental and ecological factors, endogenous hormones, sowing, fertilizer and other production management measures. For the above occurring mechanism, the proposed prevention measures are as follows: preferably germplasm, sowing, rational fertilization, exogenous hormones, shading, reed head cut off, cut fall the leaves, reasonable planting density and other agronomic measures can reduce its bolting rate. And the early bolting problem focused for future research are mentioned, including: development of authentic ingredients cultivation technical specifications, screened early bolting hormone inhibitor, in-depth study of the mechanism of early bolting, reveal its early bolting gene and key ecological factors. The research of vernalization gene, gene regulation of flowering early bolting will solve the problem of quality and production reducing fundamentally caused by the early bolting.

[Cloning and expression analysis of 4-diphosphocytidyl-2-C-methyl-D-erythritol kinase gene in Cinnamomum camphora].

The 4-diphosphocytidyl-2-C-methyl-D-erythritol kinase was the fourth key enzymes in plant terpenoid biosynthesis pathway of methyl erythritol phosphate pathway(MEP). According to the study of Cinnamomum camphora transcriptome data,we abtained the 4-diphosphocytidyl-2-C-methyl-D-erythritol kinase gene using RT-PCR,and named CcCMK1,then deposited it in GeneBank(Accession number： Ku376098).Bioinformatics analysis showed the open reading frame (ORF) of the CcCMK1 was 1 212 bp.The putative protein encoded 403 amino acids,and its molecular weight was 44.46 kDa and theoretically isoelectric point was 4.99.Transmembrane structure analysis showed that there was no transmembrane structure. Signal peptide analysis showed that it was a non secretory protein, and there was no signal peptide. The subcellular localization showed that the chloroplast was located in the chloroplast.Analysis of the expression of CcCMK1 gene in five chemotypes of C. camphora using Real-time PCR showed its expression level was highest in C. longepaniculatum, and the lowest in Borneol camphor.This research provided a basis for characterizing the key enzyme genes of terpenoid biosynthetic pathway in C. camphora.

[Effect of herb residue on growth and active ingredient content of licorice].

Traditional Chinese medicine industry product a lot of herb residue. Herb residue was treated as household waste. This treatment leads to environmental pollution and resource waste. For this case, we study the effect of different herb residues on the growth and active ingredient content of Licorice by random control experiment. Our results showed that the effects of different herb residues were difference. Atractylodes macrocephala residue and Forsythia suspense residue had the stronger effect and the effect of A. macrocephala residue was inferior to the effect of F. suspense residue. A.macrocephala residue significantly improved the shoot biomass banch number, leaf number, root diameter and root biomass by 0.53-1.81 fold. A. macrocephala residue also increased the glycyrrhizic acid content of root by 1.54 fold. F. suspense residue significantly improved the shoot biomass,branch number, root diameter and root biomass by 0.43-1.13 fold. Four kind herb residues all improved the shoot biomass by 0.43-1.81fold. So, the authors recommand to considered that we can apply A. macrocephala residue and F. suspense residue in Licorice cultivation.