Cloning，Bioinformatics and Expression Analysis of UGDH1 Gene in Glycyrrhiza uralensis / 中国实验方剂学杂志

Objective:To clone uridine diphosphate （UDP）-glucose dehydrogenase （<italic>UGDH</italic>） gene of <italic>Glycyrrhiza uralensis</italic> and analyze its bioinformatics and expression. Method:Total RNA was extracted from roots， stems， and leaves of 6-week-old seedlings of <italic>G. uralensis</italic>， the complementary deoxyribonucleic acid （cDNA） sequence of <italic>GuUGDH</italic>1 gene （Gu was short for <italic>G. uralensis</italic>） was cloned by reverse transcription polymerase chain reaction （RT-PCR）， then sequencing and bioinformatic analysis were performed， and the specificity of the tissue was analyzed by real-time fluorescence quantitative PCR （Real-time PCR）. Result:The open reading frame（ORF）of <italic>GuUGDH</italic>1 gene was 1 443 bp in length and encoded 480 amino acid residues （GenBank accession number of MT968993）. Bioinformatics analysis showed that GuUGDH1 was a stable acidic hydrophilic protein with a relative molecular weight of 53.056 kDa， an isoelectric point of 5.89， no signal peptide and no transmembrane helix， and all of them were outside the membrane. There were three typical conserved domains， which belonged to the UDP-glucose/guanosine diphosphate （GDP）-mannose dehydrogenase family. Phylogenetic analysis showed that the <italic>GuUGDH</italic>1 gene was closely related to <italic>Glycine max</italic> and <italic>Spatholobus suberectus</italic>. The results of Real-time PCR showed that the expression of <italic>GuUGDH</italic>1 gene could be detected in the roots， stems， and leaves of 6-week-old seedlings of <italic>G. uralensis</italic>， and the expression level in the roots was significantly higher than that in the stems and leaves. Conclusion:In this study， the <italic>UGDH</italic>1 gene of <italic>G. uralensis</italic> was cloned and its protein sequence characteristics were systematically analyzed， which can provide theoretical basis for further research on the catalytic function of UGDH1 protein.

The diversity of endophytic fungi in endangered medicinal material Daphnes Cortex as determined by high-throughput sequencing technology / 药学学报

In order to provide a scientific basis for the establishment of a Daphnes Cortex medicinal material fungus library and the screening of endophytic fungi that promote the growth of Daphnes Cortex and increase the content of daphnetin, we used Illumina high-throughput testing technology to analyze 9 Daphnes Cortex samples from Gansu and Shanxi provinces. A total of 632 766 valid sequences were obtained, including 348 OTUs, 4 phyla, 20 classes, 48 orders, 108 families, 154 genera, and 208 species. The sum of the first 3 fungal genera account for more than 65% of the total abundance, with the highest reaching 98.4%. Alternaria and Phoma are the main genuses of Daphne giraldii Nitsche, and Altemaria is the dominant genus. The endophytic fungi community of Daphnes Cortex is rich in diversity, and the order of fungal diversity in different producing areas is: Gangu County > Wutai County > Tanchang County.

New qualities control strategy of Scutellaria baicalensis by isothermal titration calorimetry / 中国中药杂志

It has been focused on that there will be precipitates when decoction of Scutellariat Radix mixed with Coptidis Rhizoma. Precipitation was derived from interaction between acidic and basic compounds. This study was based on the interaction between active ingredients after compatibility, strived to explore whether it was feasible to judge the qualities of different Scutellariat Radix by isothermal titration calorimetry (ITC), build a new method established to characterize the qualities of traditional Chinese medicine by taking a series of active ingredients as index. We selected Scutellariat Radix (including three batches of different Scutellariat Radix bought from market and immature Scutellariat Radix which usually was used as adulterant) in different batches as the samples. First, we used ITC to determine the binding heat of the reactions between berberine and the decoctions of different Scutellariat Radix. The test showed that the binding heat of berberine titrated Scutellariat Radix was Scutellariat Radix A (－317.20 μJ), Scutellariat Radix B (－292.83 μJ), Scutellariat Radix C (－208.95 μJ) and immature Scutellariat Radix (－21.53 μJ), respectively. We chose deionized water titrated by berberine (2.51 μJ) as control. The heat change of berberine titrated immature Scutellariat Radix was much less than berberine titrated Scutellariat Radix. Then we determined the absorbance of different decoctions of Scutellariat Radix by UV Spectrophotometry on the maximum absorption wavelength, and the result is： Scutellariat Radix A (0.372), Scutellariat Radix B (0.333), Scutellariat Radix C (0.272), immature Scutellariat Radix (0.124). The absorbance of immature Scutellariat Radix was also less than Scutellariat Radix. The result of ITC assay was corresponded to UV spectrophotometry test. In conclusion, ITC could be used to characterize the quality of Scutellariat Radix. The new method to characterize the qualities of traditional Chinese medicine by taking a kind of active ingredients as index building by ITC was simple, scientific and feasible.

Resource situation investigation and analysis about endangered Chinese herbal medicine Daphnes Cortex / 中国中药杂志

In order to figure out the status and distribution of the wild and cultivated resources of traditional Chinese medicine Daphnes Cortex, its suitable habitat and endangering factors were analyzed to provide the basis for its rational use, protection and cultivation.Our research group tooka resources survey in Shanxi, Gansu, Sichuan and Qinghai provinces, which include 23 counties. Investigation and sampling investigation combined with interview were carried out. The total reserve of resources was estimated through route-quadrat method in combination with the vegetation and soil-type map area method. The results indicated that there was no obvious change between the present distribution ranges of the wild Daphnes Cortex and its historical records, but the density of the population has undergone major changes. The wild reserves resources has declined seriously, even on the verge of exhaustion in some regions. According to the survey results, the current total reserve of the wild Daphnes Cortex in the four provinces was no more than 600 tons. Simultaneously, we only found the cultivated resource in a mountain at an altitude of about 2 800 m in Kang county of Gansu province, which cropping scope was about 33 000 m². The cultivated resource can't provide medicinal products at present, because their growing period is too short to have curative effect. Destructive excavation and the longer growth cycle result in a sharp decline of the wild resources reserves, even to the point of extinction. Artificial cultivation of product will become the main source of medicinal resources in the future. Therefore, we must protect its suitable habitat, formulate rational harvesting policy, strengthen the supervision of government departments, collect and establish the germplasm nursery and seed bank. On the basis, we must carry out studies into seed-selecting and breeding as well as rapid propagation and growth technology at once.

Effect of exogenous brassinolide on morphological characters and contents of seven chemical constituents of Glycyrrhiza uralensis / 中国中药杂志

The transplants of the two-year-old Glycyrrhiza uralensis were subjected to four concentration of brassinolide (BR 0.1, 0.4, 0.7, 1.0 mg•L⁻¹) in July. The morphological characters ( plant height, stem diameter, nodes number, internode length and root length , root thick, root fresh weight and root dry weight ) were measured and seven kinds of chemical constituents (glycyrrhizic acid, liquiritin, isoliquiritin, liquiritigenin, isoliquiritigenin, liquiritin apioside, isoliquiritin apioside) were determined by HPLC with the aim of increasing sinter output and improving quality of G. uralensis. Then the long-term dynamic changes of these morphological characters and chemical compositions' content were analyzed. The results showed that morphological characters of plant height, stem diameter, root length , root thick, root fresh weight and root dry weight increased remarkably with the 0.7 mg•L⁻¹ BR stimulating 2 months later,the increase rates were： 15.09%,6.15%,16.52%,8.46%,21.90%,29.41%, respectively. The content of glycyrrhizic acid, liquiritin, isoliquiritin, liquiritigenin, liquiritin apioside, isoliquiritin apioside were increased 20.16%,45.31%,53.56%,27.66%,23.54%,8.46% with the 0.7 mg•L⁻¹ BR stimulating 2 months later. The best effects were achieved in 2 months after brassinolide stimulating. The conclusions prove that morphological characters and the main chemical constituents accumulation of G. uralensis could be effected by exogenous BR stimulation in certain case.

Resource situation investigation about Rheum tanguticum and its sustainable utilization analysis in main production area of China / 中国中药杂志

This study was conducted to investigate the wild and cultivated resource situation of Rheum tanguticum in main production area of China, estimate its reserves, and put forward the feasible approach for the sustainable utilization of R. tanguticum. On the basis of the literature data about R. tanguticum, conbined with interview, investigation and sampling investigation, the total reserve of resources is estimated using the route-quadrat method and the vegetation and soil-type map area method proposed by our research group. The results indicate that there is no obvious change between the present distribution ranges of the wild R. tanguticum and its historical records, but its population density has changed clearly. The reserve of the wild R. tanguticum has seriously declined in lots of place, even faced the exhaustion in some regions. According to the investigation, the resource reserve of the wild R. tanguticum is no more than 5 000 t, and the cultivated is about 1 607 t. The resource reserve of the wild R. tanguticum is nearly depleted, and this suggests that the wild R. tanguticum should be enrolled in the protection plant list, and the cultivated will become the main resource of Rhubarb in the future. So it is extremely neccessary to collect and protect the germplasm resource of R. tanguticum, establish the germplasm nursery and repository, and conduct breeding research on those bases.

Procedure of seed quality testing and seed grading standard of Prunus humilis / 中国中药杂志

So far there exists no corresponding quality test procedures and grading standards for the seed of Prunus humilis, which is one of the important source of base of semen pruni. Therefor we set up test procedures that are adapt to characteristics of the P. humilis seed through the study of the test of sampling, seed purity, thousand-grain weight, seed moisture, seed viability and germination percentage. 50 cases of seed specimens of P. humilis tested. The related data were analyzed by cluster analysis. Through this research, the seed quality test procedure was developed, and the seed quality grading standard was formulated. The seed quality of each grade should meet the following requirements: for first grade seeds, germination percentage ≥ 68%, thousand-grain weight 383 g, purity ≥ 93%, seed moisture ≤ 5%; for second grade seeds, germination percentage ≥ 26%, thousand-grain weight ≥ 266 g, purity ≥ 73%, seed moisture ≤9%; for third grade seeds, germination percentage ≥ 10%, purity ≥ 50%, thousand-grain weight ≥ 08 g, seed moisture ≤ 13%.

Wild-tending techniques study on Glycyrrhiza uralensis--effect of irrigation and rhizome length on survival ratio, yield and quality / 中国中药杂志

This research aimed at studying the effects of irrigation and rhizome length on the survival of ratio, yield and quality of Glycyrrhiza uralensis in wild tending condition. Employed the split-block design to carry out the field experiment, sampled with the quadrat method to measured the relative growth indexes and to estimate the yield, used the HPLC (high performance liquid chromatog- raphy ) method to measure the glycyrrhizin in the rhizome and adventitious root of the G. uralensis in this study. The quantity of the adventitious roots and the survival ratio were increased significantly as the length of the rhizome increased (P < 0.01), but the length of the rhizome had no remarkable effect on the content of glycyrrhizin. The average content of the glycyrrhizin in the adventitious root and rhizome could reach 3.03% and 2.12% after 3-year wild tending, respectively, and this results indicated that the quality of the glycyrrhiza using this method was much better than that from cultured glycyrrhiza with the reproducing method of seeding. so using the rhizome as reproductive material to produce the glycyrrhiza under the wild tending condition could get the high quality glycyrrhiza quick- ly and steadily, this phenomenon could be explained by the Hypothesis of synthetic inertia of the medicinal components from the wild material of G. uralensis. But the maximum yield with this method was just more than 945 kg x hm(-2) in this study. So the further work of how to increase the yield in the practical application with the method found in this study need to be done in the next research.