[Rapid extraction of DNA from Chinese medicinal products by alkaline lysis].

The study is aimed to explore a rapid method to extract DNA from fried Chinese medicinal products. The alkaline lysis buffer was made of sodium hydroxide, 1% PVP and 1% TritonX-100 and Tris-HCl solution was neutralized, through heat cracking and neutralization two step to extract DNA from processed and prepared products of traditional Chinese medicine. Then universal primes were used to amplify PCR products for fired Chinese medicinal materials. The results indicated the optimized alkaline lysis method for extracting DNA is quick and easy. Extracting of the different processed Sophora japonica of DNA concentration was (420.61 ± 123.91) g x L(-1). Using 5% Chelex-100 resin purification can improve the DNA concentration. Our results showed that the optimized alkaline lysis method is suitable for Chinese medicinal materials for quickly DNA extraction.

[Distribution and morphological variation of germplasm resource of Sophora alopecuroides].

OBJECTIVE: To analyze the genetic difference of biological characters on germplasm resources of Sophora alopecuroides. METHOD: Twenty-three populations of S. alopecuroides from Ningxia, Gansu, Qinghai, Xinjiang and Inner Mongolian were used to analyze the seed size, 1 000-grain weight, and germination characteristics and so on. RESULT: It showed that there were significant differences in seed size, 1 000-grain weight and the vitality of seeds. The biggest seed of S. alopecuroides was 4.7 mm x 3.5 mm, and the smallest was 3.8 mm x 2.9 mm, and the 1 000-grain weight was 15-26 g. Results of seeds vitality in 8 populations indicated that the highest vitality of seeds were No. 103 and No. 122. The germination index was 36.51 and 36.24 respectively, and the vitality index was 1 323.49 and 1 274.56. The coefficient of variation in seed traits exceeded 10% except the seed size. CONCLUSION: There are some differences and different heredity background in various S. alopecuroides germplasm resources.

The complete chloroplast genome sequence of the medicinal plant Sophora tonkinensis.

Sophora tonkinensis belongs to genus Sophora of the Fabaceae family. It is mainly distributed in the ridge and peak regions of limestone areas in western China and has high medicinal value and important ecological functions. Wild populations of S. tonkinensis are in danger and need urgent conservation. Furthermore, wild S. tonkinensis resources are very limited relative to the needs of the market, and many adulterants are present on the market. Therefore, a method for authenticating S. tonkinensis and its adulterants at the molecular level is needed. Chloroplast genomes are valuable sources of genetic markers for phylogenetic analyses, genetic diversity evaluation, and plant molecular identification. In this study, we report the complete chloroplast genome of S. tonkinensis. The circular complete chloroplast genome was 154,644 bp in length, containing an 85,810 bp long single-copy (LSC) region, an 18,321 bp short single-copy (SSC) region and two inverted repeat (IR) regions of 50,513 bp. The S. tonkinensis chloroplast genome comprised 129 genes, including 83 protein-coding genes, 38 transfer RNA (tRNA) genes, and 8 ribosomal RNA (rRNA) genes. The structure, gene order and guanine and cytosine (GC) content of the S. tonkinensis chloroplast genome were similar to those of the Sophora alopecuroides and Sophora flavescens chloroplast genomes. A total of 1,760 simple sequence repeats (SSRs) were identified in the chloroplast genome of S. tonkinensis, and most of them (93.1%) were mononucleotides. Moreover, the identified SSRs were mainly distributed in the LSC region, accounting for 60% of the total number of SSRs, while 316 (18%) and 383 (22%) were located in the SSC and IR regions, respectively. Only one complete copy of the rpl2 gene was present at the LSC/IRB boundary, while another copy was absent from the IRA region because of the incomplete structure caused by IR region expansion and contraction. The phylogenetic analysis placed S. tonkinensis in Papilionoideae, sister to S. flavescens, and the genera Sophora and Ammopiptanthus were closely related. The complete genome sequencing and chloroplast genome comparative analysis of S. tonkinensis and its closely related species presented in this paper will help formulate effective conservation and management strategies as well as molecular identification approaches for this important medicinal plant.

Complete chloroplast genome of Sophora alopecuroides (Papilionoideae): molecular structures, comparative genome analysis and phylogenetic analysis.

Sophora alopecuroides belongs to the genus Sophora of the family Papilionoideae. It is mainly distributed in the desert and semidesert areas of northern China, and has high medicinal value and ecological function. Previous studies have reported the chemical composition and ecological functions of S. alopecuroides. However, only a few reports are available on the genomic information of S. alopecuroides, especially the chloroplast genome, which greatly limits the study of the evolutionary relationship between other species of Papilionoideae. Here, we report the complete chloroplast genome of S. alopecuroides. The size of the chloroplast genome is 155,207 bp, and the GC content is 36.44%. The S. alopecuroides chloroplast genome consists of 132 genes, including 83 protein-coding genes, 41 transfer RNA (tRNA) genes,and eight ribosomal RNA (rRNA) genes. Phylogenetic analysis revealed the taxonomic position of S. alopecuroides in Papilionoideae, and the genus Sophora and the genus Ammopiptanthus were highly related. Comparative genomics analysis revealed the gene rearrangement in the evolution of S. alopecuroides. The comparison between S. alopecuroides and the species of the Papilionoideae identified a novel 23 kb inversion between the trnC-GCA and trnF-GAA which occurred before the divergence of Sophora and Ammopiptanthus of Thermopsideae. This study provided an essential data for the understanding of phylogenetic status of S. alopecuroides.

Flavonoids rather than alkaloids as the diagnostic constituents to distinguish Sophorae Flavescentis Radix from Sophorae Tonkinensis Radix et Rhizoma: an HPLC fingerprint study.

Sophorae Flavescentis Radix (Sophora flavescens Ait., SFR) and Sophorae Tonkinensis Radix et Rhizoma (S. tonkinensis Gapnep., STR) are two commonly used traditional Chinese medicines from Sophora (Leguminosae) plants, which are believed to possess similar bioactive components with entirely different clinical applications. In order to find out the characteristic chemical constituents potentially leading to the unique medicinal properties claimed for each of the two closely related TCMs, an HPLC fingerprint method was developed for analyses of the alkaloid and flavonoid constituents of SFR and STR, respectively, which were further evaluated and compared through similarity calculation and hierarchical clustering analysis (HCA). The results from the present study showed that the alkaloid fingerprints of the two herbs were similar, with many components co-existing in both drugs and various batches of samples from different species being mixed together in the HCA dendrogram. However, their flavonoid constituents were totally different with specific fingerprints being yielded for each herb, and further HCA analysis showed that the tested samples could almost be clearly divided into two groups based on their origins of species. The results from the present study indicated that the flavonoid constituents could serve as the differentially diagnostic constituents of SFR and STR and might potentially attributed to their distinct therapeutic effects.

Electron Ionization Mass Spectrometry-based Metabolomics Studies of Sophoraflavescens can Identify the Geographical Origin of Root Samples.

An electron ionization mass spectrometry (EI-MS)-based metabolomic approach was applied to Sophora flavescens to identify the geographical origin of each sample. The score plot from principal component analysis using the EI-MS data showed that Japanese S. flavescens samples tended to cluster away from Chinese S. flavescens samples. Statistical techniques showed that ions arising from kurarinol and kushenol H, which we previously identified as marker molecules for Japanese S. flavescens, were characteristic of Japanese S. flavescens. Therefore, metabolomics based on EI-MS data is a valuable tool for confirming the geographical origins of S. flavescens samples. The results suggest that EI-MS-based metabolomics is suitable for the quality control of traditional medicines containing many components.

Effects of Total Alkaloids of Sophora alopecuroides on Biofilm Formation in Staphylococcus epidermidis.

Staphylococcus epidermidis (S. epidermidis) is an opportunistic pathogen with low pathogenicity and a cause of the repeated outbreak of bovine mastitis in veterinary clinical settings. In this report, a biofilm model of S. epidermidis was generated and the minimal inhibitory concentration (MIC) and sub-MIC (SMIC) on bacterial cultures were assessed for the following agents: total alkaloids of Sophora alopecuroides (TASA), ciprofloxacin (CIP), and erythromycin (ERY). The formation and characteristic parameters of biofilm were analyzed in terms of XTT assay, silver staining, and confocal laser scanning microscope (CLSM). Results showed that a sub-MIC of TASA could inhibit 50% biofilm of bacterial activity, while 250-fold MIC of CIP and ERY MICs only inhibited 50% and 47% of biofilm formation, respectively. All three agents could inhibit the biofilm formation at an early stage, but TASA showed a better inhibitory effect on the late stage of biofilm thickening. A morphological analysis using CLSM further confirmed the destruction of biofilm by these agents. These results thus suggest that TASA has an inhibitory effect on biofilm formation of clinic S. epidermidis, which may be a potential agent warranted for further study on the treatment prevention of infection related to S. epidermidis in veterinary clinic.

TEK, local perceptions of risk, and diversity of management practices of Agave inaequidens in Michoacán, Mexico.

BACKGROUND: Mescal production is the main economic activity associated to agaves in Mexico, which involves 53 species mostly harvested from forests. The increasing mescal demand has influenced risk in both agave populations and mescal production, but other social and ecological factors also intervene. We hypothesized that the greater the risk the greater the complexity of management responses; otherwise, the greater the probability of populations' depletion. We analysed this hypothesis by examining the diversity of risk conditions and management practices of Agave inaequidens in the state of Michoacán, in central-western Mexico. METHODS: We studied five communities of Michoacán, documenting through 41 semi-structured interviews the use forms, risk perception, number of agaves annually extracted, and the management practices. Using a matrix with social-ecological and technological data analyzed by PCA, we evaluated similarities of management contexts. A data matrix with information on risk of agave populations, and other about management practices were analysed also through CCA and PCA. The scores of the first principal components were considered as indexes of risk and management complexity, respectively. A regression analysis of these indexes evaluated their relation. RESULTS: We recorded 34 different uses of A. inaequidens, the most important being mescal production (mentioned by 76.1 % of people interviewed). Nearly 12.5 % of people practice only gathering, but others mentioned the following practices: Selective let standing of agaves for seed production (20 %); in situ transplanting of saplings; seed propagation in nurseries and saplings transplanting to forest (10 %); suckers transplanting (7.5 %); seed dispersal in forests; banning (5 %); enhancing of agave growth by removing tree canopies (2.5 %); transplanting from the wild to live fences (45 %); intensive plantations (35 %). The highest vulnerability of agave populations was identified in communities where risk is not counteracted by management. In two communities we identified the highest risk (annual extraction from 4,353 to 6,557 agaves), but different actions counteracting such risk. CONCLUSIONS: Interchange of knowledge and management experiences developed by handlers is crucial for the regional conservation, recovering, and sustainable management of A. inaequidens populations.

Analysing the floral elements of the lost tree of Easter Island: a morphometric comparison between the remaining ex-situ lines of the endemic extinct species Sophora toromiro.

Sophora toromiro (Phil) Skottsb. is a species that has been extinct in its natural habitat Easter Island (Rapa Nui) for over 50 years. However, seed collections carried out before its extinction have allowed its persistence ex-situ in different botanical gardens and private collections around the world. The progenies of these diverse collections have been classified in different lines, most of them exhibiting high similarity as corroborated by molecular markers. In spite of this resemblance observed between the different lines, one of them (Titze) has dissimilar floral elements, thus generating doubts regarding its species classification. The floral elements (wing, standard and keel) belonging to three different S. toromiro lines and two related species were analyzed using geometric morphometrics. This method was applied in order to quantify the floral shape variation of the standard, wing, and keel between the different lines and control species. Geometric morphometrics analyses were able to distinguish the floral elements at both intra (lines) and inter-specific levels. The present results are on line with the cumulative evidence that supports the Titze line as not being a proper member of the S. toromiro species, but probably a hybridization product or even another species of the Edwardsia section. The reintroduction programs of S. toromiro should consider this information when assessing the authenticity and origin of the lines that will be used to repopulate the island.

Classification using NMR-based metabolomics of Sophora flavescens grown in Japan and China.

We demonstrate that NMR-based metabolomics can be used to identify the country of growth (Japan or China) of Sophora flavescens plants. Principle Component Analysis (PCA) conducted on extracts of S. flavescens grown in China provided data distinct from that of extracts of plants grown in Japan. Loading plot analysis showed signals characteristic of Japanese S. flavescens. NMR analyses showed these signals to be due to kurarinol (1) and kushenol H (2). These compounds were confirmed by HPLC analysis to be distinctive markers for Japanese S. flavescens.

[Structural characteristics of Sophora moorcroftiana community on wind-sandy land in middle reaches of Yaluzangbu River].

Based on quadrat investigation and hierarchical cluster analysis, this paper studied the structural characteristics of Sophora moorcroftiana community at its different restoration succession stages on the wind-sandy land in middle reaches of Yaluzangbu River. The plant community on the wind-sandy land could be divided into 6 types, i.e., S. moorcroftiana community, Orinus thoroldii community, S. moorcroftiana < O. thoroldii community, S. moorcroftiana-O. thoroldii community, Artemisia weiibyii+S. moorcroftiana community, and S. moorcroftiana+A. weiibyii community. The restoration succession began with the sand-fixing S. moorcroftiana distributed on moving dune, followed by O. thoroldii invasion, and then, S. moorcroftiana-O. thoroldii community formed gradually. Another restoration succession approach began with O. thoroldii community, S. moorcroftiana occurred then, and finally, S. moorcroftiana-O, thoroldii community formed. On semi-fixed dune, A. weiibyi could invade the community, and S. moorcroftiana+A. weiibyi community formed. The climax community on the wind-sandy land was the shrub community dominated by S. moorcroftiana. With increasing slope grade, the coverage and height of S. moorcroftiana community tended to decrease. Biological crust had definite improvement effect on the soil physical and chemical properties. At the places with biological crust, surface soil water content was significantly higher, compared with that at the places with other restoration succession stages of S. moorcroftiana community. The crown diameter, plant height, and the sprouting branch number of S. moorcroftiana community were the highest on moving dune, followed by on fixed dune, and on semi-fixed dune. The dead branch number at different restoration succession stages indicated that S. moorcroftiana population had a declining trend after the sand was fixed.

[Dynamics of Sophora davidii population in a hilly region closed for thirty years in Loess Plateau].

Taking wild grass ground as the reference, this paper studied the dynamics of Sophora davidii population on the sunny, semi-sunny and semi-shady slopes of hilly region in Loess Plateau after 30 years closed for reforestation, and analyzed the age structure, spatial distribution pattern, life table, and survival curve of the population. The results showed that S. dvidii population in the region belonged to progressive type, with more young and less old individuals. Among the three habits, semi-sunny slope had the highest population density. The survival curve of S. dvidii population was approached to Deevey 11 type. The population pattern was of clustered distribution, with the aggregating intensity increased with age. Water and light were the main environmental factors affecting the recovery and development of S. davidii community. The better recovery of S. davidii population suggested that the measures of closing the land for reforestation were efficient.