Research on Pollution-free and Technical Regulations of Andrographis Paniculata / 世界科学技术-中医药现代化

Andrographis paniculata is a traditional Chinese herbal medicine with a wide cultivation in south China. The market demand has been increasing steadily in recent years. However, there are problems such as disordered production, pesticide residues and heavy metal content in the process of Andrographis paniculata production. Pollution- free cultivation is an effective strategy to solve this problem, and it is also the development direction of the Chinese herbal medicine planting industry at this stage. In order to guide the cultivation of pollution-free and high-quality Andrographis paniculata, this study established a pollution- free fine cultivation technology system of Andrographis paniculata including precise cultivation and site selection. In addition, soil improvement, scientific pollution-free planting mode and high- efficiency integrated pest control technology help solve problems such as pesticide residues and heavy metal content. This paper provides a reference for the pollution-free and fine cultivation of Andrographis paniculata.

Research on Pollution-free and Technical Regulations of Paeonia suffruticosa and Paeonia lactiflora / 世界科学技术-中医药现代化

Moutan Cortex and Paeonia Radix are also traditional Chinese herbal medicines. The market is in great demand. However, the market sales of Moutan Cortex and Paeonia Radix have problems such as pesticide residues, excessive heavy metals and degraded quality, which affect their safety and effectiveness. The establishment of a pollution-free and planting system can effectively reduce the pesticide residue and heavy metal content and improve the quality of the medicinal materials while ensuring the production of Moutan Cortex and Paeonia Radix. The article uses field data which based on P.suffruticosa and P.lactiflora cultivation, combined with growth habits and distribution of origin, this research has developed a pollution-free and planting technology system for P.suffruticosa and P.lactiflora. The system includes planting base regionalization, field management, fertilization, pest control and so on, which provides reference for P.suffruticosa and P.lactiflora.

Discussion on Pollution-free and Technical Regulations of Rabdosia rubescens (Hemsl.) Hara and Rabdosia serra (Maxim.) Hara / 世界科学技术-中医药现代化

Rabdosia (Bl.) Hassk is a medicinal plant rich in enantiomers such as 15α-acetoxyl-6, 11α-epoxy-6α-hydroxy-20-oxo-6, 7-seco-ent-kaur-16-en-1, 7-olide and 15α-dihydroxy-6β-methoxy-6, 7-seco-6, 20-epoxy-1, 7-olide-ent-kaur-16-ene. It has high medicinal value and huge market potential, but the yield from wild Rabdosia (Bl.) Hassk species cannot meet the needs of marketization. Carrying out pollution-free cultivation of Rabdosia rubescens (Hemsl.) Hara and Rabdosia serra (Maxim.) Hara has become the most direct and effective measure to solve this problem in order to ensure the sustainable use of the resources of Rabdosia. This paper summarized a set of non- pollution cultivation system of R. rubescens and R. serra to achieve the"safe, effective, stable and controllable"target.

Effects of Abiotic Elicitors MeJA and SA on Alkaloids Accumulation and Related Enzymes Metabolism inPinellia ternata Suspension Cell Cultures / 中国中医药信息杂志

Objective To study the effects of abiotic elicitors methyl jasmonate (MeJA) and salicylic acid (SA) on the alkaloids accumulation and related enzymes metabolism inPinellia ternata suspension cell cultures. Methods Using the leaf petioles-derived suspension cell cultures as the study object, the culture duration, concentrations of MeJA and SA were determined to get the optimal alkaloids accumulation, and the activities of metabolic enzymes IMP dehydrogenase and sAMP synthase were also measured.Results A 9-fold of dried biomass and a 3-fold of alkaloids accumulation were observed inP. ternata suspension cell cultures after culture for 21 d. Both MeJA and SA could significantly promote the accumulation of alkaloids inP. ternata suspension cells. 150 μmol/L MeJA enhanced alkaloids content (4.7 mg/gDW) by 3.6 folds in comparison with control group, whereas 50 μmol/L SA showed a 2.5-fold increase. Meanwhile, 100 μmol/L MeJA and 50 μmol/L SA promoted the increase in IMP dehydrogenase activity by 3.0 and 3.7 fold respectively, and 150 μmol/L MeJA and 100 μmol/L SA showed the increase by 2.6 and 4.4 fold respectively.Conclusion Proper adding exogenous MeJA and SA can promote the accumulation of alkaloids inPinellia ternata suspension cell cultures.

EMS mutation of suspension cells and selection of high temperature tolerant mutants from Pinellia ternata / 中国中药杂志

<p><b>OBJECTIVE</b>To determine the optimal concentration and processing time of EMS mutation for suspension cells from Pinellia ternata.</p><p><b>METHOD</b>Under four EMS concentration gradients (0.1% , 0.2%, 0.4%, 0.6%) and three processing time gradients (0.5, 1.0, 2.0 h), the suspension cells of P. ternata were mutagenized.</p><p><b>RESULT AND CONCLUSION</b>The results showed that the survival rate was significantly different under the different concentrations of EMS and the different processing time. In the same processing time, the EMS concentrations were increased, but the suspension cells survival rate decreased gradually. The optimum EMS concentration for the mutagenesis was 0.4% and the best processing time was 1 hour.</p>

Comparison of different methods for isolating total RNA from bulblet of Fritillaria anhuiensis / 中国中药杂志

<p><b>OBJECTIVE</b>To optimize a simple and effective method for total RNA extraction from bulblet of Fritillaria anhuiensis.</p><p><b>METHOD</b>Four methods, i. e. guanidine isothiocyanate, bentonite, modified SDS/phenol and the RNAiso plus, were used to extract total RNA from bulblet of F. anhuiensis. Then the results of the extraction were compared and analyzed by electrophoresis detection and RT-PCR verification.</p><p><b>RESULT</b>The total RNA extracted by bentonite method were clear and no dispersion, the integrity of the RNA was well, and there was no obvious contamination with DNA and other impurities, was suitable for RT-PCR test.</p><p><b>CONCLUSION</b>The bentonite method is quick, economic, and efficient for total RNA extraction from bulblet of F. anhuiensis.</p>

Effect of low temperature on cell synchronization division in suspension culture cells of Pinellia ternata / 中国中药杂志

<p><b>OBJECTIVE</b>To study the effect of low temperature on cell synchronization division in suspension culture cells of Pinellia ternata, in order to find a proper condition for cell synchronization.</p><p><b>METHOD</b>The loosened calli of P. ternata were used as materials. The effect of low temperature on cell synchronization division was studied and researched in suspension culture cells by the method of orthogonal experiment, the obtained experimental data was compared and analyzed through the analysis of range and analysis of variance.</p><p><b>RESULT</b>After the low temperature treatment, the cell division index was increased in various degrees, among the total results, when the temperature was set at 8 degrees C for 24 hours and then recovered for 36 hours, the effect was the best, the mitotic index was significantly increased to 11.3%.</p><p><b>CONCLUSION</b>The low temperature could obviously influence the cell synchronization division in suspension culture cells of P. ternata, and the mitotic index was efficiently increased.</p>

Differential display of mRNA from microtubers of Pinellia ternata in vitro / 中国中药杂志

<p><b>OBJECTIVE</b>To study mRNA differential expression from microtubes of Pinellia ternata in vitro, and give more information of the molecular mechanism in the formation of microtubers.</p><p><b>METHOD</b>DDRT-PCR was used to identify the expressed gene fragments related to the microtubers development of P. ternata in vitro.</p><p><b>RESULTS AND CONCLUSION</b>Fifteen cDNA fragments differentially expressed in the induction of microtubers of P. ternata in vitro were identified and characterized by using mRNA differential display DDRT-PCR. The deduced amino-acid sequences of six fragments of cDNA showed no significant homology with ESTs and genes in the Genbank databases, and they could be new cDNA fragments. However, the remaining three showed significant homologies with sequences encoding components of eukaryotic translation initiation factor 3 subunit H, MADS-box protein and ethylene signal transcription factor, respectively. Their differential expression patterns were confirmed by semi-quantitative RT-PCR analysis. And the expression level of the induction of microtubers of P. ternata in vitro was different.</p>

Study on preservation of Fritillaria anhuiensis by vitrification in vitro / 中国中药杂志

<p><b>OBJECTIVE</b>To establish cryopreservation system of shoot-tips from Fritillaria anhuiensis.</p><p><b>METHOD</b>Taking vitrification as system of cryopreservation, the shoot tips with length 2-3 mm were precultured in MS medium enriched with 0.4 mol x L(-1) sucrose for 3 d. They were treated for 20 min with 60% PVS2 at 25 degrees C, and then subjected to ice-cooled vitrification solution for 60 min and transferred to 2 mL cryotubes with fresh vitrification solution (PVS2) and plunged into liquid nitrogen. After rapid thawing in 40 degrees C water bath for 1 min, shoot-tips were expelled into MS medium containing 1.2 mol x L(-1) sucrose for 20 min. Further recovery and growth took place on regeneration medium in the dark at 25 degrees C for 2 weeks, and then with light/dark cycle of 12/12 h. The genetic integrity of cryopreserved shoot-tips was identified through products of PCR with arbitrary primers.</p><p><b>RESULT AND CONCLUSION</b>The highest survival rates of shoot-tips reached 79.9% by vitrification, and the regeneration rates were 52.3%. No changes were found between treated materials and untreated materials in genomic DNA.</p>

Study on hydroxyproline tolerance of different cultivars of Chrysanthemum morifolium in vitro / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate hydroxyproline tolerance of Chrysanthemum morifolium plantlets included " Boju", "Huaiju", "Chuju", "Gongju" and "Hangju",and provide references basis for excellent cultivar and breeding of Ch. morifolium.</p><p><b>METHOD</b>Plantlets in vitro from five kinds of Ch. morifolium were inoculated on medium added with different concentrations of hydroxyproline. Free proline in leaves from plantlets was determined, then the damage index and survival rate were compared.</p><p><b>RESULT AND CONCLUSION</b>The results showed that hydroxyproline tolerance of " Boju" and "Huaiju" were superior, the survival rates and free proline of them were higher, but the damage index was inferior. The hydroxyproline tolerance of "Hangju" was the worst, and the survival rate was minimum. The survival rate of "Chuju" and "Gongju" was between "Boju" and " Hangju", and the hydroxyproline tolerance of them was also medium.</p>

Development and verification of Chinese dietary exposure evaluation model software / 中华预防医学杂志

<p><b>OBJECTIVE</b>To develop the dietary exposure evaluation model software accredited of Chinese intellectual property rights and to verify the rationality and accuracy of the results from the probabilistic model in Chinese dietary exposure evaluation model software according to international standards.</p><p><b>METHODS</b>The software of SAS was used to build various evaluation model based on the data from Chinese dietary survey and the chemical compound in food surveillance and to design an operation interface. The results from probabilistic dietary exposure model for children 2 - 7 years old were compared with that from duplicate portion study of 2-7 years children dietary exposure in Jinhu, Jiangsu province in order to analyze the rationality of model. The results from probabilistic model of dietary exposure were compared with the results from @Risk software to verify the correction of the probabilistic model by using the same data of randomly selected 10 000 study subjects from national dietary survey. While, the mean drift was used as an internal index to illustrate the accuracy of the computation.</p><p><b>RESULTS</b>Chinese dietary exposure evaluation software was developed successfully. On the rationality, the results from probabilistic model were lower than that from the point estimation (e.g., cucumber: the result of point estimation of acephate was 4.78 microg x kg(-1) x d(-1), while the results of probabilistic model which was 0.39 microg x kg(-1) x d(-1)). Meanwhile the results from probabilistic model were higher than the results of duplicate portion study (on the P95, the result of probabilistic model of Pb exposure in children was 11.08 microg x kg(-1) x d(-1), while the results of duplicate portion study was 5.75 microg x kg(-1) x d(-1)). On accuracy, the results from @Risk and the probabilistic model were highly consistent (on the P95, the result of probabilistic assessment of acephate diet exposure was 4.27 microg x kg(-1) x d(-1), while the results of duplicate portion study was 4.24 microg x kg(-1) x d(-1)), and the mean drift was of random distribution, the drift region varied from 0.05% to 11.9%.</p><p><b>CONCLUSION</b>The results computed by the software of Chinese dietary exposure evaluation model are reliable and reasonable, which is a meaningful step to improve the dietary exposure evaluation technique in China.</p>

Effects of Pinellia ternata extracts on inhibiting of oviposition and ovicidal action against Plutella xylostella / 中国中药杂志

<p><b>OBJECTIVE</b>To study the bioactivities of the extracts from Pinellia ternata against Plutella xylostella.</p><p><b>METHOD</b>The active material in P. ternata was Soxhlet extracted with petroleum ether, ethyl acetate, ethanol successively. The bioactivities of the extracts were determined.</p><p><b>RESULT</b>Results indicated that all extracts using three solvents showed inhibition of oviposition and ovicidal action activities, but the activity of ethanol extract was the strongest. The higher concentration, the more obviously the effect on the pest was. 24 h after treatment with 100 g x L(-1) ethanol extract in no-choice tests and choice tests, the deterrent rates were 85.13%, 73.38%, respectively. Furthermore, treatment with 100 g x L(-1) ethanol extract after 5 d could effectively reduced the percentage of eggs incubation, ovicidal rate could reach 62.40%.</p><p><b>CONCLUSION</b>These results suggest that extracts of P. ternata could be used to control pest.</p>

Changes of total RNA and mRNA differential expressions in leaves of Pinellia ternata under high temperature stress / 中国中药杂志

<p><b>OBJECTIVE</b>To study the total RNA and mRNA differential expression in leaves of Pinellia ternata under high temperature, provide more information of the molecular mechanism of the sprout tumble.</p><p><b>METHOD</b>The total RNA and mRNA differential expression in leaves of P. ternata at different stress time was analyzed.</p><p><b>RESULT</b>The results showed that the trend of total RNA content was divided into three descending stages and two ascending stages, the total RNA content was the highest at 0, 6 h, but it was the lowest at and 42 h, as well as when the sprout tumbled. The differential display showed that the polymorphism and type of bands of the sample at 6 h were similar to those at 0 h. But the bands numbers at other time were far less than those at 0, 6 h. And there were some different mRNA differential expression bands between the different samples.</p><p><b>CONCLUSION</b>In the process of the sprout tumble caused by high temperature stress, the RNA and mRNA differential expression in leaves of P. ternata changed.</p>

Variation of endogenous hormones in formation of microtuber of Dioscorea opposite in vitro / 中国中药杂志

<p><b>OBJECTIVE</b>Through analysis of variation and function of 5 main endogenous hormones in the formation of microtuber of Dioscorea opposite in vitro to explore the physiological and biochemical mechanism of microtuber development.</p><p><b>METHOD</b>When microtubers were induced on MS + 6-BA 1.5 mg x L(-1) + NAA 1.5 mg x L(-1) + sucrose 5% medium, the endogenous hormones were isolated during different formation stages of microtubers, then purified and detected with enzyme-linked immunosorbent assays (ELISA).</p><p><b>RESULT</b>The results showed that GA3 slightly decreased in initial period, rose suddenly 20 days later, and than decreased. IAA showed a dropping tendency in the total course, ABA and ZR increased in a long period, dropped at last. JA continuously rose and never dropped, GA3 and ABA and the ratio of GA3 and JA varied obviously.</p><p><b>CONCLUSION</b>IAA, ABA, JA , ZR and GA3 play an important role in controlling formation of microtubers in D. opposite in vitro.</p>

Changes of photosynthesis parameters and chlorophyll fluorescence around sprout tumble of Pinellia ternata under high temperature stress / 中国中药杂志

<p><b>OBJECTIVE</b>To study the change of photosynthesis and chlorophyll fluorescence characteristics of Pinellia ternata around sprout tumble.</p><p><b>METHOD</b>Tubers of P. ternata were cultured firstly at (25 +/- 1) degrees C for certain days, and then they were coerced under 37 degrees C stress in the same artificial climate boxes. The chlorophyll contents, photosynthesis and chlorophyll fluorescence during different stages of high temperature stress were measured.</p><p><b>RESULT AND CONCLUSION</b>After under high temperature stress, the chlorophyll contents, the ratio of chlorophyll a/b, photosynthetic rate (Pn), stomatal conductance (Gs), transpiration rate (Tr), maximum fluorescence (Fm), quantum yield of PS II (phi (PS II)), intrinsic photochemical efficiency of PS II (Fv/Fm) and the chlorophyll photochemical quenching (qP) decreased, but the intercellular CO2 concentration (Ci), minimal fluorescence (F0), the chlorophyll nonphotochemical quenching (NPQ) increased.</p>

Research on callus induction and plantlet regeneration of Prunella vulgaris / 中国中药杂志

<p><b>OBJECTIVE</b>To study the conditions of callus induction and plantlet regeneration of Prunella vulgaris.</p><p><b>METHOD</b>By using the orthogonal experiment design, different explants, sucrose, plant growth substances and their ratio for callus induction and differentiation were optimized.</p><p><b>RESULT AND CONCLUSION</b>The inductivity of leaves was the highest, followed by stems, but callus of the leafstalks could not be induced. The main factor of affecting the formation of callus was 6-BA. The optimal medium for callus induction was MS + 6-BA 3.0 mg x L(-1) + NAA 0.1 mg x L(-1) + 2,4-D 0.5 mg x L(-1) + sucrose 3%. The optimal medium for callus differentiation was 1/2 MS + 6-BA 3.25 mg x L(-1) + NAA 1.25 mg x L(-1) + sucrose 2%. The optimal medium for rooting was MS + IBA 1.0 mg x L(-1) + sucrose 3%.</p>

Variation of endogenous hormones in formation of microtubers of Pinellia ternata in vitro / 中国中药杂志

<p><b>OBJECTIVE</b>Through analysis of variation and function of five endogenous hormones (IAA, GA3, ABA, ZR and JA) in the formation of microtubers of Pinellia ternata in vitro to explore the physiological and biochemical mechanism of tubers' development.</p><p><b>METHOD</b>The endogenous hormones of microtubers were isolated in different stages of tubers expansion when the leaf stalks of P. ternata were cultured on MS + 6-BA 0.5 mg x L(-1) + NAA 0.1 mg x L(-1) + sucrose 5% medium to induce the formation of microtubers, then them and stems were detected with Enzyme-linked Immunosorbent Assays (ELISA).</p><p><b>RESULT</b>The results showed that the content of IAA, ABA, JA and ZR in the intumescentia period of microtubers were rapidly rising. The curve of GA3 appeared the type of anti-S.</p><p><b>CONCLUSION</b>IAA, ABA, JA, ZR and GA3 played an important role in controlling formation of microtubers in P. ternata.</p>

Callus induction and cultivation of Phyllanthus urinaria / 中草药

Objective To establish culture method for Phyllanthus urinaria. Methods To study the possible effective factors of culture condition by comparing with different explants,sucrose,plant growth substance,and its ratio. Results The inductivity of stem was the highest about 55.56%,but callus of leaves could not be induced.6-BA was the most important factor on callus induction,followed by 2,4-D NAA and sucrose. Conclusion The optimal medium to induce and subculture is MS+2,4-D 0.5 mg/L+6-BA 1.0 mg/L+NAA 0.1 mg/L+sucrose 10 g/L.