Transcription factor DcbZIPs regulate secondary metabolism in Dendrobium catenatum during cold stress.

Cold stress seriously affects plant development and secondary metabolism. The basic region/leucine zipper (bZIP) is one of the largest transcription factor (TFs) family and widely involved in plant cold stress response. However, the function of bZIP in Dendrobium catenatum has not been well-documented. Cold inhibited the growth of D. catenatum and increased total polysaccharide and alkaloid contents in stems. Here, 62 DcbZIP genes were identified in D. catenatum, which were divided into 13 subfamilies. Among them, 58 DcbZIPs responded to cold stress, which were selected based on the transcriptome database produced from cold-treated D. catenatum seedlings. Specifically, the expression of DcbZIP3/6/28 was highly induced by cold treatment in leaves or stems. Gene sequence analysis indicated that DcbZIP3/6/28 contains the bZIP conserved domain and is localized to the cell nucleus. Co-expression networks showed that DcbZIP6 was significantly negatively correlated with PAL2 (palmitoyl-CoA), which is involved in flavonoid metabolism. Moreover, DcbZIP28 has significant negative correlations with various metabolism-related genes in the polysaccharide metabolic pathway, including PFKA1 (6-phosphofructokinase), ALDO2 (aldose-6-phosphate reductase) and SCRK5 (fructokinase). These results implied that DcbZIP6 or DcbZIP28 are mainly involved in flavonoid or polysaccharide metabolism. Overall, these findings provide new insights into the roles of the DcbZIP gene family in secondary metabolism in D. catenatum under cold stress.

[Current researches and prospects of seedling propagation and cultivation modes of Jinxianlian].

In the wake of on-the-spot investigation into Chinese major production bases in Fujian, Zhejiang, Jiangxi, Taiwan, Guangxi and Yunnan provinces, and based on relevant literature, the paper systematically elaborates the current researches of botanical origin, major cultivation type, seedling propagation technique as well as cultivation mode of Jinxianlian. The way of seedling breeding mainly includes aseptic seed culture, in vitro propagation, artificial seed and bioreactor propagation, etc. And the planting model mainly includes protected cultivation modes, bionic wild cultivation modes and pot cultivation modes, etc. Further discussions have also been conducted to tackle significant problems existing in the production process of Jinxianlian, based on personal studies of the authors. It has made considerable contributions for the betterment of Jinxianlian's development and improvement.

[Current status and sustainable development countermeasures of Anoectochilus roxburghii].

The current status of the domestic manufacturing and sales markets of Anoectochilus roxburghii were investigated and analyzed in the study. Some problems in the A. roxburghii industry were revealed and a variety of sustainable development countermeasures were also proposed. The main problems of A. roxburghii industry are the lack of protection for wild resources, the lag in the speed of variety breeding, the insufficient research on the quality systems, the low level of industry and product innovation capability, as well as the relatively low market cognition and brand competence. Therefore, strengthening the protection for breeding resources, establishing a dynamic monitoring system, promoting the variety breeding, constructing a propagation system for improved varieties, enhancing the quality of medicinal herbs, accelerating the adjustment of product structure, upgrading the industry technology, strengthening brand competence and expanding the market, will be the effective methods to realize the sustainable development of A. roxburghii industry.

Application of response surface methodology to optimise supercritical carbon dioxide extraction of volatile compounds from Crocus sativus.

BACKGROUND: Crocus sativus has been used as a traditional Chinese medicine for a long time. The volatile compounds of C. sativus appear biologically active and may act as antioxidants as well as anticonvulsants, antidepressants and antitumour agents. In order to obtain the highest possible yield of essential oils from C. sativus, response surface methodology was employed to optimise the conditions of supercritical fluid carbon dioxide extraction of the volatile compounds from C. sativus. Four factorswere investigated: temperature, pressure, extraction time and carbon dioxide flow rate. Furthermore, the chemical compositions of the volatile compounds extracted by supercritical fluid extraction were compared with those obtained by hydro-distillation and Soxhlet extraction. RESULTS: The optimum extraction conditions were found to be: optimised temperature 44.9°C, pressure 34.9 MPa, extraction time 150.2 min and CO2 flow rate 10.1 L h⁻¹. Under these conditions, the mean extraction yield was 10.94 g kg⁻¹. The volatile compounds extracted by supercritical fluid extraction and Soxhlet extraction contained a large amount of unsaturated fatty acids. CONCLUSION: Response surface methodology was successfully applied for supercritical fluid CO2 extraction optimisation of the volatile compounds from C. sativus. The study showed that pressure and CO2 flow rate had significant effect on volatile compounds yield produced by supercritical fluid extraction. This study is beneficial for the further research operating on a large scale.

[Effects of different transplanting conditions on survival rate and growing status of Anoictochilus roxburghii plantlets].

The growing status of Anoectochilus roxburghii seedling was observed and the survival rate of seedlings, height, stem diameter and plant fresh weight under the conditions of different transplanting substrate compositions, planting density, shading rate were measured. The results showed that the effects of different transplanting substrates, planting densities, shading rates and nutrient solutions on the growing status of A. roxburghii plantlets varied greatly. A. roxburghii plantlets demonstrated a high survival rate and better growing status under the Following conditions: the ratio of peat and river sand as 2: 1, the planting density as 3 cm x 3 cm, the shading rate as 70%, and the nutrient solution as 1/4MS. The findings of the study provide a solid technical solution for the artificial cultivation of A. roxburghii plantlets.

[Influence of seedling grade on plant growth, yield and quality of Anoectochilus roxburghii].

The morphological index of the seedlings including the plants height, the ground diameter, the leaf amounts, the fresh weight of the whole plant and the ratio of height to diameter was measured and the principal components were analyzed so as to determine the grading index, and stepwise cluster analysis was applied for clustering analysis. Pot experiments were used to measure the indicators of plant growth and development, the yield and the quality. The results showed that the height and ground diameter were determined as the quality indicators of the seedlings grading and the standard quality grading of seedlings of Anoectochilus roxburghii was initially set up, different seeding plants influenced the plants growth and the yield. The ground diameter of the class I was larger than that of the class II and III, so as the yield. The seedling grading had no obvious effect on the internal quality of medicinal materials. The results of the study provide the basis for standard cultivation of A. roxburghii.

Gut microbiota and transcriptome profiling revealed the protective effect of aqueous extract of Tetrastigma hemsleyanum leaves on ulcerative colitis in mice.

Tetrastigma hemsleyanum, a traditional Chinese medicinal plant, possesses various biological activities, including anti-inflammatory and immunomodulatory functions. The purpose of this study was to determine the alleviating effect of the water extract of Tetrastigma hemsleyanum leaves (THLW) on ulcerative colitis (UC) and its relationship with gut microbiota. The administration of THLW significantly decreased the severity of dextran sulfate sodium (DSS)-induced intestinal damage, as demonstrated by the stabilization of body weight and colon length, and decreased disease activity index (DAI) and histological scores. THLW also decreased NF-κB protein expression in colon tissues and reduced the serum levels of IL-6, IL-1ß, and TNF-α. Further co-housing experiment confirmed that the anti-UC effect of THLW was possibly by regulating the structure and composition of gut microbiota, including increasing the abundance of Oscillospiraceae, Prevotellaceae and Corynebacterium. Additionally, the expression of genes related to inflammation and immunity was also regulated by THLW treatment as evidenced by transcriptome analysis. These results suggested that the protective effect of THLW on DSS-induced colitis was mediated by alleviating inflammation and modulating the microbiota composition. This work proved the potent protective effects of THLW treatment on colitis and may have potential for UC relief.

[Simultaneous determination of four alisols in Rhizoma Alismatis by RP-HPLC].

OBJECTIVE: To develop an HPLC method for the determination of alisol A, alisol F, alisol A 24-acetate and alisol B 23-acetate in the rhizome of Alisma orientalis. METHOD: The separation was performed on a Shim-Pack CLC-ODS a C18 column (6 mm x 150 mm, 5 microm) eluted with the mobile phases of acetonitrile (A)-water (B) in gradient elution. The absorbance was monitored at 210 nm. Orthogonal test was adopted to optimize the extraction conditions of the four alisols. RESULT: The correlation coefficients of the four alisols were higher than 0.999 and the average recoveries were 99.23%, 96.67%, 97.30% and 99.61%, respectively. All the RSDs were less than 3%. CONCLUSION: The validation data demonstrated that the method was accurate and repeatable, and can be used to measure the four alisols in Rhizoma Alismatis.

Identification of anti-tumor components from toad venom.

Secretion of granular glands from the skin of amphibians is a fascinating resource of active substances, particularly for cancer therapy in clinical practice of Traditional Chinese Medicine. A variety of anti-tumor peptides have been isolated from different toads and frogs; however, no anti-tumor peptides are reported in toad venom of Bufo gargarizans. Firstly, soluble fraction from fresh toad venom (FTV) was compared with that from dried toad venom (DTV), using HPLC analysis. It was revealed that FTV has a different HPLC chromatography compared with DTV. Soluble fraction of FTV is more toxic to SH-SY5Y cells than that of DTV, as evaluated by MTT assay. Secondly, it was identified that protein components from soluble fractions of FTV and DTV possess different patterns by SDS-PAGE analysis, and proteins from FTV are also more toxic than that from DTV. Thirdly, an immobilized basic fibroblast growth factor (bFGF) affinity column was used to isolate bFGF-binding components from soluble fraction of FTV, and it was identified that bFGF-binding components prohibited bFGF-dependent neurite growth of SH-SY5Y cells. Finally, it was identified that bFGF-binding components activated apoptosis via upregulation of caspase-3 and caspase-8 expression in SH-SY5Y cells. These data suggest that FTV contains active components that interact with bFGF and activate apoptosis in SH-SY5Y cells.

Effects of shade treatments on photosynthetic characteristics, chloroplast ultrastructure, and physiology of Anoectochilus roxburghii.

Anoectochilus roxburghii was grown under different shade treatments-50%, 30%, 20%, and 5% of natural irradiance-to evaluate its photosynthetic characteristics, chloroplast ultrastructure, and physiology. The highest net photosynthetic rates and stomatal conductance were observed under 30% irradiance, followed in descending order by 20%, 5%, and 50% treatments. As irradiance decreased from 50% to 30%, electron transport rate and photochemical quenching increased, while non-photochemical quenching indexes declined. Reductions in irradiance significantly increased Chl a and Chl b contents and decreased Chl a/b ratios. Chloroplast ultrastructure generally displayed the best development in leaves subjected to 30% irradiance. Under 50% irradiance, leaf protein content remained relatively stable during the first 20 days of treatment, and then increased rapidly. The highest peroxidase and superoxide dismutase levels, and the lowest catalase activities, were observed in plants subjected to the 50% irradiance treatment. Soluble sugar and malondialdehyde contents were positively correlated with irradiance levels. Modulation of chloroplast development, accomplished by increasing the number of thylakoids and grana containing photosynthetic pigments, is an important shade tolerance mechanism in A. roxburghii.