[Diversity of agronomic traits of Paris polyphylla var. chinensis and their correlation with rhizome yield and active compositions].

The supply deficiency of crude medicinal plant of Paris polyphylla var. chinensis has become a bottleneck for related medicinal industry. An important approach to increase herbal production is to breed high-yield cultivated variety, which characterized ideal plant morphology. In the present study, we collected 99 wild germplasm resources of P. polyphylla and then measured their 12 main agronomic traits and contents of polyphyllin Ⅶ,Ⅵ,Ⅱ,Ⅰ. Followed analyses were used to characterize those traits and explore the potential connection with herbal yield or quality. The results showed that: ①There was ample morphological diversity in wild P. polyphylla, whose variation of agronomic traits reduced according to followed order: content of polyphyllin, weight of dry rhizome, petiole length, stem length, petal length, pedicel length, sepal length, leaf width, leaf length, sepal width, leaf number, stamen number, petal number. ② Most of those traits were significantly correlated to each other and generally represented the characterization of photosynthetic organs or reproductive organ. ③The total content of polyphyllin Ⅶ,Ⅵ,Ⅱ,Ⅰvaried between 0.02% and 0.87% and averagedat 0.13%, which showed no significant correlation with any agronomic trait. ④Plant breeders should play more attention on those germplasm resources with large leaves, large sepals and high stem.

Metabolomics approach reveals annual metabolic variation in roots of Cyathula officinalis Kuan based on gas chromatography-mass spectrum.

BACKGROUND: Herbal quality is strongly influenced by harvest time. It is therefore one of crucial factors that should be well respected by herbal producers when optimizing cultivation techniques, so that to obtain herbal products of high quality. In this work, we paid attention on one of common used Chinese herbals, Cyathula officinalis Kuan. According to previous studies, its quality may be related with growth years because of the variation of several main bioactive components in different growth years. However, information about the whole chemical composition is still scarce, which may jointly determine the herbal quality. METHODS: Cyathula officinalis samples were collected in 1-4 growth years after sowing. To obtain a global insight on chemical profile of herbs, we applied a metabolomics approach based on gas chromatography-mass spectrum. Analysis of variance, principal component analysis, partial least squares discriminant analysis and hierarchical cluster analysis were combined to explore the significant difference in different growth years. RESULTS: 166 metabolites were identified by using gas chromatography-mass spectrum method. 63 metabolites showed significant change in different growth years in terms of analysis of variance. Those metabolites then were grouped into 4 classes by hierarchical cluster analysis, characterizing the samples of different growth ages. Samples harvested in the earliest years (1-2) were obviously differ with the latest years (3-4) as reported by principal component analysis. Further, partial least squares discriminant analysis revealed the detail difference in each growth year. Gluconic acid, xylitol, glutaric acid, pipecolinic acid, ribonic acid, mannose, oxalic acid, digalacturonic acid, lactic acid, 2-deoxyerythritol, acetol, 3-hydroxybutyric acid, citramalic acid, N-carbamylglutamate, and cellobiose are the main 15 discrimination metabolites between different growth years. CONCLUSION: Harvest time should be well considered when producing C. officinalis. In order to boost the consistency of herbal quality, C. officinalis is recommended to harvest in 4th growth year. The method of GC-MS combined with multivariate analysis was a powerful tool to evaluate the herbal quality.

[Cloning, subcellular lacalization and expression analysis of chloroplast-targeted Obg gene in Cyathula officinalis].

According to ObgC gene sequences from Cyathula officinalis genomic data, the specific primers were designed, and a full-length CoObgC cDNA (2 226 bp) was obtained by polymerase chain reaction (PCR) and rapid amplification of cDNA ends (RACE) methord. Sequence alignment showed that CoObgC gene contained a 1 818 bp open reading frame (ORF) encoding 605 amino acids. Sequence analysis predicted that molecular weight of CoObgC protein was about 66.39 kDa, the academic isoelectric point was 5.35, and the protein was stable protein. Then multiple sequence alignment was applied to construct phylogenetic tree. The real-time fluorescence quantification PCR (RT-qPCR) demonstrated that a high expression level in leaf, followed by root and flower, the low transcription was in stem. The recombinant vector pCABIA2300-CoObgC was constructed and introduced into tobacco epidermal cells by agrobacterium-mediated transformation, green fluorescence was tested and targeted to chloroplast under a laser scanning confocal microscope. These findings will be helpful to lay a foundation for studying the structure and function of CoObgC gene, and elucidating C. officinalis molecular biology experiment.

[Effects of polysaccharides isolated from Panax japonicus on immunosuppression mice].

OBJECTIVE: To investigate the immunomodulatory effects of polysaccharides from Panax japonicus on immunosuppressed mice. METHODS: Immunosuppressed murine model were established in Kuming mice induced by cyclophosphamide (Cy) and splenic indexes were evaluated. The proliferation activities of splenic lymphocyte were assayed by MTT and levels of serum hemolysin were confirmed by spectrophotometry. The antibody-secreting capabilities of splenocytes were assayed by quan-titative haemolysis of SRBC (QHS). The percentage of NK cells were analysed by flow cytometry (FCM). RESULTS: Polysaccharides of Panax japonicus significantly increased the splenic indexes, promoted the proliferation of splenic lymphocytes, enhanced QHS reaction, elevated the production of serum hemolysin and percentages of peripheral blood NK cells in Cy-induced immunosuppressed mice. CONCLUSION: Panax japonicus polysaccharides can recover immune function in the Cy-induced immunosuppressed mice.