Molecular cloning and functional characterization of an isoflavone glucosyltransferase from Pueraria thomsonii.

Pueraria thomsonii has long been used in traditional Chinese medicine. Isoflavonoids are the principle pharmacologically active components, which are primarily observed as glycosyl-conjugates and accumulate in P. thomsonii roots. However, the molecular mechanisms underlying the glycosylation processes in (iso)flavonoid biosynthesis have not been thoroughly elucidated. In the current study, an O-glucosyltransferase (PtUGT8) was identified in the medicinal plant P. thomsonii from RNA-seq database. Biochemical assays of the recombinant PtUGT8 showed that it was able to glycosylate chalcone (isoliquiritigenin) at the 4-OH position and glycosylate isoflavones (daidzein, formononetin, and genistein) at the 7-OH or 4'-OH position, exhibiting no enzyme activity to flavonones (liquiritigenin and narigenin) in vitro. The identification of PtUGT8 may provide a useful enzyme catalyst for efficient biotransformation of isoflavones and other natural products for food or pharmacological applications.

[Analysis of genomic SSRs in Pueraria lobata and P. thomsonii and establishment of DNA identity card for different germplasms of P. thomsonii of Jiangxi province].

The type and frequency of simple sequence repeats( SSRs) in the genomes was investigated using the DNA sequence data of Pueraria lobata and P. thomsonii. Based on these SSRs,20 pairs of SSR primers were designed and 5 high polymorphism primer pairs were selected to analyze genetic diversity of 9 cultivars of P. thomsonii in Jiangxi province. The results showed that the 5 pairs of primers could generate 16 polymorphic alleles bands. The average polymorphism information content( PIC) of each SSR primer pair was 0. 600 7.According to the genetic similarity coefficients,the 9 cultivars of P. thomsonii can be classified into 6 germplasms. This study established DNA identity cards with 5 pairs of SSR primers for different germplasm resources of P. thomsonii in Jiangxi province,which provided reference information for the selection of fine germplasms of P. thomsonii and the theoretical basis for the study of Dao-di herbs.

[Growth rings in roots of seven Sect. Paeonia species and its application on identification of Paeoniae Radix Rubra].

The growth years of medicinal materials are closely related to their quality, and "Herb-chronology" has been used to determine the growth years of perennial dicotyledonous plants in recent years. On the basis of conventional paraffin section and freehand section, the anatomical study on roots of seven Sect. Paeonia species and main roots of cultivated Paeonia lactiflora was conducted in this paper. The results showed that, there existed some differences in microstructure of the seven species such as P. lactiflora, P. obovata, P. veitchii, P. mairei, P. anomala, P. sinjiangensis and P. anomala var. intermedia, and this could be used to distinguish different species. In the roots of seven Sect. Paeonia species, distinct growth rings were formed because that the different diameters or density of xylem vessels in the secondary xylem formed clusters and arranged interrupted rings in tangential direction. There were growth rings in the main roots of P. lactiflora cultivated 1-4 years in Siping, Jilin, which were all consistent with their growth years. Due to the similar growth characteristics between wild Sect. Paeonia species and cultivated P. lactiflora, the growth rings can provide a basis for the age identification and lay the foundation for the quality evaluation of Paeoniae Radix Rubra.

[Geographical distribution of Chinese herbalists：causes of four distribution centers about Huizhou, Shanghai, Hangzhou, Suzhou].

Based on the "Zhong Guo Ben Cao Yao Ji Kao", Chinese herbalists in past dynasties were counted and analyzed by their living period,numbers and native places. Combined with GIS, the geographical distribution and the formation causes of the four distribution centers of herbalists in past dynasties were discussed. The results showed that, there was a greater difference between the numbers of herbalists in different periods of time, which achieved to the peak in the Ming and Qing Dynasties. In addition, the distribution of herbalists in past dynasties characterized east more and west less, forming the distribution areas centered by Huizhou, Shanghai, Hangzhou and Suzhou. Besides, the geographical distributions of Chinese herbalists showed an obvious southward trend since the Song Dynasty.

Rapid identification of growth years and profiling of bioactive ingredients in Astragalus membranaceus var. mongholicus (Huangqi) roots from Hunyuan, Shanxi.

BACKGROUND: The content of medicinal bioactive constituents in huangqi is affected by plant age. In this study, we devised a quick and convenient method for determining the age of huangqi, which was cultivated in Hunyuan County (Shanxi Province). METHODS: 1, 2, 3, 4, 5, 8, 10 growth years huangqi had 38 samples, all samples were collected separately. The growth rings in these samples were observed after making paraffin section and freehand-section. The relationship between growth rings and its growth years was analyzed by SPSS 19.0 software. Histochemical localization of total flavones and saponins in huangqi was determined by color reactions. The concentration of four flavonoids and two saponins in the roots of huangqi of different ages and different organizational structure (normal roots and rotten heart roots) were determined by HPLC-DAD and HPLC-ELSD. The results were analyzed by SPSS 19.0 software. RESULTS: All huangqi samples had clear growth rings, and the statistical result about growth rings (X) and growth years (Y) showed significant correlation (r = 1, P = 0.000). The calibration curves of these six ingredients showed good linearity respectively, with significant correlation. All relative standard deviations (RSDs) of precision, recovery, repeatability, and stability experiments were less than 2%. Roots of 5-year-old plants contained the highest concentrations of total flavonoids and saponins. Saponin concentrations increased toward the center of the roots, whereas the four flavonoids showed an opposite trend in tissue distribution. CONCLUSION: The growth year of huangqi (Hunyuan County, Shanxi Province) could be determined soon and conveniently by naked eyes after staining phloroglucinol-HCl solution on freehand section. The content of saponins and flavonoids in rotten heart root and the surrounding normal tissues were affected by the formation and the extent of rotten heart.

Identification of four Aconitum species used as "Caowu" in herbal markets by 3D reconstruction and microstructural comparison.

Authentication is the first priority when evaluating the quality of Chinese herbal medicines, particularly highly toxic medicines. The most commonly used authentication methods are morphological identification and microscopic identification. Unfortunately, these methods could not effectively evaluate some herbs with complex interior structures, such as root of Aconitum species with a circular conical shape and an interior structure with successive changes. Defining the part that should be selected as the standard plays an essential role in accurate microscopic identification. In this study, we first present a visual 3D model of Aconitum carmichaeli Debx. constructed obtained from microscopic analysis of serial sections. Based on this model, we concluded that the point of largest root diameter should be used as the standard for comparison and identification. The interior structure at this point is reproducible and its shape and appearance can easily be used to distinguish among species. We also report details of the interior structures of parts not shown in the 3D model, such as stone cells and cortical thickness. To demonstrate the usefulness of the results from the 3D model, we have distinguished the microscopic structures, at their largest segments, of the other three Aconitum species used for local habitat species of Caowu. This work provides the basis for resolution of some debate regarding the microstructural differences among these species. Thus, we conclude that the 3D model composed of serial sections has enabled the selection of a standard cross-section that will enable the accurate identification of Aconitum species in Chinese medicine.

[Investigation on Main Cultivation of Astragalus membranaceus var. mongholicus and Commercial Specifications Grades].

OBJECTIVE: To investigate the main cultivation areas of Astragalus membranaceus var. mongholicus, and to clarify its present cultivation status and commercial specification grades. METHODS: Astragalus membranaceus var. mongholicus cultivated in Hunyuan and Ying County of Shanxi, Longxi, Weiyuan and Min County of Gansu and Wuchuan, Guyang and Chifeng of Inner Mongolia were investigated, including their ecological environment, cultivation history and present situation, harvesting and processing as well as commercial specification grades. RESULTS: The cultivation methods of Astragalus membranaceus var. mongholicus changed greatly in the main cultivation areas. In Gansu and Inner Mongolia, the current major cultivation method was 2 - 3 years of seedling transplantation method, while the 3 - 5 years of seed-directly-sowed method in Inner Mongolia had been almost abandoned, and the 6 - 8 years of mountain direct seeding method was still in application in Hunyuan county of Shanxi province. On the other hand, the harvest method of Astragalus membranaceus var. mongholicus had been gradually changed from manual harvesting in ancient times to the current mechanical harvesting although the latter method did more damage to environment of cultivation areas and the resource of Astragalus membranaceus var. mongholicus in regions Hunyuan county of Shanxi. Through investigating the processing methods of Astragalus membranaceus var. mongholicus in different cultivation areas, it was found that processing method varied from area to area, and no unified specifications had been formulated on processing methods in market. In addition, the current processing method was too complicated in processing the slices of prepared Astragali Radix and most were self processing by farmers themselves, creating market disorders. Moreover, the Astragali Radix products were basically without grading classification but sold as bulk cargo. CONCLUSION: It is urgent to unify the specifications and grades of Astragali Radix product, and standardize its processing methods.

[Microscopic anatomy of abnormal structure in root tuber of Pueraria lobata].

Puerariae Lobatae Radix, also known as Gegen, is a root derived from Pueraria lobata. Based on field investigation and the developmental anatomy of root tuber, we have elucidated the relationship between the growth of root tuber and the anomalous structure. The results of analysis showed that the root system of P. lobata was developed from seed and adventitious root and there existed root tuber, adventitious root and conductive root according to morphology and function. The root tuber was developed from adventitious root, its secondary structure conformed to the secondary structure of dicotyledon's root. With the development of root, the secondary phloem of root tuber appeared abnormal vascular tissue, which was distributed like ring in the outside of secondary vascular tissue. The root tuber might have 4-6 concentric circular permutation abnormal vascular tissuelobate, and was formed by the internal development of abnormal vascular tissue. The xylem and phloem of abnormal vascular tissue were the main body of the root tuber. The results reveal the abnormal anatomical structure development of P. lobata, also provides the theoretical basis for reasonable harvest medicinal parts and promoting sustainable utilization of resources of P. lobata.

[Reasearch on evolution and transition of processing method of fuzi in ancient and modern times].

Fuzi is a medicine used for rescuing from collapse by restoring yang as well as a famous toxic traditional Chinese medicine. In order to ensure the efficacy and safe medication, Fuzi has mostly been applied after being processed. There have been different Fuzi processing methods recorded by doctors of previous generations. Besides, there have also been differences in Fuzi processing methods recorded in modern pharmacopeia and ancient medical books. In this study, the authors traced back to medical books between the Han Dynasty and the period of Republic of China, and summarized Fuzi processing methods collected in ancient and modern literatures. According to the results, Fuzi processing methods and using methods have changed along with the evolution of dynasties, with differences in ancient and modern processing methods. Before the Tang Dynasty, Fuzi had been mostly processed and soaked. From Tang to Ming Dynasties, Fuzi had been mostly processed, soaked and stir-fried. During the Qing Dynasty, Fuzi had been mostly soaked and boiled. In the modem times, Fuzi is mostly processed by being boiled and soaked. Before the Tang Dynasty, a whole piece of Fuzi herbs or their fragments had been applied in medicines; Whereas their fragments are primarily used in the modern times. Because different processing methods have great impacts on the toxicity of Fuzi, it is suggested to study Fuzi processing methods.