An exploration of mechanism of high quality and yield of Gastrodia elata Bl. f. glauca by the isolation, identification, and evaluation of Mycena.

Gastrodia elata Bl. f. glauca is an important traditional Chinese medicinal plant. The yield and quality of Gastrodia elata Bl. have significantly decreased due to multigenerational asexual reproduction. Therefore, it is necessary to have sexual reproduction of Gastrodia elata Bl. to supplement the market supply. Seeds of G. elata Bl. have no endosperm, and their sexual reproduction depends on the nutrients provided by the embryo cells infected by Mycena fungi to complete seed germination. However, Mycena fungi are small and have many species, and not all Mycena fungi can promote the germination of G. elata Bl. seeds. Therefore, it is of great significance to isolate and identify suitable germination fungi and explore the mechanism for improving the production performance and yield, and quality of G. elata Bl. Six closely related Mycena isolates, JFGL-01, JFGL-02, JFGL-03, JFGL-04, JFGL-05, and JFGL-06, were isolated from the leaves and protocorms of G. elata Bl. f. glauca and were identified as Mycena purpureofusca. The mycelial state and number of germinating protocorms were used as indicators to preferentially select Mycena fungi, and it was concluded that JFGL-06 had the best mycelial state and ability to germinate G. elata Bl. seeds. Finally, a mechanism to increase the yield of G. elata Bl. was explored by comparing the changes in nutrient elements and microbial diversity in the soil around G. elata Bl. with different strains. JFGL-06 proved to be an excellent Mycena fungal strain suitable for G. elata Bl. f. glauca. Compared with the commercial strain, JFGL-06 significantly increased the C, N, Na, Mg, S, Cl, K, Ca, and Fe contents of the soil surrounding the protocorms of G. elata Bl. f. glauca. JFGL-06 improved the composition, diversity, and metabolic function of the surrounding soil microbial community of G. elata Bl. f. glauca protocorms at the phylum, class, and genus levels, significantly increased the relative abundance of bacteria such as Acidobacteria and fungi such as Trichoderma among the dominant groups, and increased the abundance of functional genes in metabolic pathways such as nucleotide metabolism and energy metabolism. There was a significant reduction in the relative abundance of bacteria, such as Actinomycetes, and fungi, such as Fusarium, in the dominant flora, and a reduced abundance of functional genes, such as amino acid metabolism and xenobiotic biodegradation and metabolism. This is the main reason why the JFGL-06 strain promoted high-quality and high-yield G. elata Bl. f. glauca in Changbai Mountain.

Transcriptome analysis reveals underlying immune response mechanism of fungal (Penicillium oxalicum) disease in Gastrodia elata Bl. f. glauca S. chow (Orchidaceae).

BACKGROUND: Gastrodia elata Bl. f. glauca S. Chow is a medicinal plant. G. elata f. glauca is unavoidably infected by pathogens in their growth process. In previous work, we have successfully isolated and identified Penicillium oxalicum from fungal diseased tubers of G. elata f. glauca. As a widespread epidemic, this fungal disease seriously affected the yield and quality of G. elata f. glauca. We speculate that the healthy G. elata F. glauca might carry resistance genes, which can resist against fungal disease. In this study, healthy and fungal diseased mature tubers of G. elata f. glauca from Changbai Mountain area were used as experimental materials to help us find potential resistance genes against the fungal disease. RESULTS: A total of 7540 differentially expressed Unigenes (DEGs) were identified (FDR < 0.01, log2FC > 2). The current study screened 10 potential resistance genes. They were attached to transcription factors (TFs) in plant hormone signal transduction pathway and plant pathogen interaction pathway, including WRKY22, GH3, TIFY/JAZ, ERF1, WRKY33, TGA. In addition, four of these genes were closely related to jasmonic acid signaling pathway. CONCLUSIONS: The immune response mechanism of fungal disease in G. elata f. glauca is a complex biological process, involving plant hormones such as ethylene, jasmonic acid, salicylic acid and disease-resistant transcription factors such as WRKY, TGA.

[Research progress on mechanism of gastrodin and p-hydroxybenzyl alcohol on central nervous system].

Gastrodin(GAS) and p-hydroxybenzyl alcohol(HBA) are extracts of dried tubers of Gastrodia elata, which is the material basis for its efficacy and belongs to phenolic compounds. Modern pharmacology studies have shown that they have significant effects on central nervous system diseases, such as insomnia, convulsions, depression, ischemic stroke, anxiety, and cognitive impairment, and these diseases are closely related to neurotransmitters and cytokines. This paper described various mechanisms of GAS and HBA monomer components on the central nervous system. They alleviate hippocampal neuronal toxicity mainly by regulating a variety of neurotransmitters, such as acetylcholine, glutamic acid(GLU), γ-aminobutyric acid(GABA), serotonin(5-HT), dopamine(DA), norepinephrine(NE), 5-indoleacetic acid(5-HIAA), high vanillic acid(HVA) and dihydroxyphenylacetic acid(DOPAC), pro-inflammatory cell growth factors, such as IL-1ß, IL-6 and TNF-α and relevant receptor functions, and exert neuropharmacological effects by effectively increasing mRNA expressions of brain neurotrophic factors, such as BDNF and GDNF, and further inhibiting the apoptosis of damaged neurons. This paper summarized various mechanisms on the central nervous system, which provides a scientific basis for the further research of the neuropharmacological mechanism of GAS and HBA and the development of new drugs and functional food.

Study on the simultaneous degradation of five pesticides by Paenibacillus polymyxa from Panax ginseng and the characteristics of their products.

The quality and safety of ginseng products were seriously affected due to the slow metabolism and long-term residual pesticides in ginseng. Microbial degradation is an effective method to degrade pesticide residues. In this study, ginseng endophytic Paenibacillus polymyxa was used to degrade pesticide residues. A method of simultaneous determination of fluazinam, BHC, PCNB, chlorpyrifos and DDT in ginseng roots and ginseng stems and leaves by GC was established. The sample was extracted with n-hexane and purified by Florisil solid phase extraction column. The limit of quantitation was 0.01 µg mL-1, the linear relationship was good (r ≥ 0.9901). 7 days after inoculated with P. polymyxa, the degradation rates of fluazinam, BHC, PCNB, chlorpyrifos, and DDT in the medium were 94.77%, 70.34%, 77.92%, 78.30%, 66.70%, respectively (P < 0.05). The safety of 5 pesticide degradation products was investigated by GC-MS. The results showed that after 7 days degradation, the main degradation products were alkanes, which are non-toxic and can't cause secondary pollution to the environment. The actual degradation results were verified by field experiments. The results indicated that after sprayed 5 times with P. polymyxa, the degradation rates of fluazinam, BHC, PCNB, chlorpyrifos and DDT in the ginseng roots were 66.07%, 46.24%, 21.05%, 72.40%, 54.21%, respectively (P < 0.05). The degradation rates in ginseng stems and leaves were 74.18%, 55.61%, 73.65%, 58.13%, 46.91%, respectively (P < 0.05). The results indicated that Paenibacillus polymyxa was an effective degradation strain of 5 pesticides.

Gut Microbiota Play an Essential Role in the Antidiabetic Effects of Rhein.

It is clear that the gut microbiota can affect host metabolism and alterations of the gut microbiota can link with metabolic disease. Rhein has been used in traditional Chinese medicine with putative antidiabetic effects. Here we show that oral administration of rhein for 6 weeks can significantly reduce fasting blood glucose (FBG) level (8.30 ± 4.52 mmol/l versus 18.89 ± 6.06 mmol/l, p < 0.01), elevate the active glucagon-like peptide 1 (GLP-1) level (22.21 ± 2.61 pmol/l verss 14.46 ± 5.22 pmol/l, p < 0.05), and increase the number of L-cells in the terminal ileum. The antidiabetic effect of rhein is abrogated in db/db mice treated with rhein in combination with broad-spectrum antibiotics. We observed that the abundance of the Bacteroidetes is increased in mice treated with rhein (0.361±0.022 versus 0.185 ± 0.055, p < 0.05,). In addition, there is no significant difference in diversity between rhein-treated groups and the controls (Shannon index: p = 0.88; Simpson index: p = 0.86). Taken together, our results indicate that modulation of the gut microbiota may play an essential role in the antidiabetic effects of rhein.

[Study on early screening of high saponin ginseng lines].

The study aims at screening the specific bands by PCR, quickly and accurately evaluating the quality of ginseng seeding, accelerating the process of ginseng breeding. Based on the correlation of genetic differences and saponin content between individuals, a pair of specific primer GC1 was screened by PCR. According to the experiment by L16 (45) orthogonal test, a PCR system most suitable for GC1 was established, which came out total 25 µL reaction system containing DNA 2.60 mg•L⁻¹, Mg²âº 1.44 mmol•L⁻¹, dNTP 0.19 mmol•L⁻¹, primer 0.32 µmol•L⁻¹ and Taq enzyme concentration 0.076 U•µL⁻¹. By comparing the saponin content and the GC1 PCR electrophoretogram of samples, the ginseng, with 1 200 bp specific band by PCR of GC1, the contents of 9 monosodium saponins and their additions were higher than others, which provided a reliable method for accelerating the process of ginseng breeding. The sequence was sequenced and 99% homologous to glycerol-3-phosphate dehydrogenase.

A transgenic ginseng vaccine for bovine viral diarrhea.

BACKGROUND: Bovine viral diarrhea virus (BVDV) infections are endemic in cattle populations worldwide and cause major economic losses. Thus, an effective vaccine is needed against the transmission of BVDV. The glycoprotein E(rns) is one of the envelope proteins of this virus and shows BVDV-related immunogenicity. Here, we report the use of Panax ginseng as an alternative production platform for the expression of glycoprotein E(rns) via Agrobacterium-mediated transformation. RESULT: Polymerase chain reaction (PCR) and reverse transcription (RT)-PCR analyses showed that pBI121-E(rns) was stably integrated into the chromosome of transformants. ELISA assay and Western blot analysis confirmed the antigenicity of plant-derived E(rns) glycoprotein. Immunogenicity was evaluated subcutaneously in deer using a soluble protein extract of dried transgenic ginseng hairy roots. Specific humoral and cell-mediated immune responses against BVDV were detected following immunization. CONCLUSION: These results demonstrated that the E(rns) glycoprotein could be expressed in ginseng hairy roots and that plant-derived glycoprotein E(rns) retained its antigenicity and immunogenicity.

Generation of the bovine viral diarrhea virus e0 protein in transgenic astragalus and its immunogenicity in sika deer.

The bovine viral diarrhea virus (BVDV), a single-stranded RNA virus, can cause fatal diarrhea syndrome, respiratory problems, and reproductive disorders in herds. Over the past few years, it has become clear that the BVDV infection rates are increasing and it is likely that an effective vaccine for BVDV will be needed. In this study, transgenic Astragalus was used as an alternative productive platform for the expression of glycoprotein E0. The immunogenicity of glycoprotein E0 expressed in transgenic Astragalus was detected in deer. The presence of pBI121-E0 was confirmed by polymerase chain reaction (PCR), transcription was verified by reverse transcription- (RT-) PCR, and recombinant protein expression was confirmed by ELISA and Western blot analyses. Deer that were immunized subcutaneously with the transgenic plant vaccine developed specific humoral and cell-mediated immune responses against BVDV. This study provides a new method for a protein with weak immunogenicity to be used as part of a transgenic plant vaccine.

Mineral elements in root of wild Saposhnikovia divaricata and its rhizosphere soil.

Mineral elements are important components of medicinal herbs, and their concentrations are affected by many factors. In this study, Ca, Mg, Na, K, Fe, Mn, Cu, and Zn concentrations in wild Saposhnikovia divaricata and its rhizosphere soil collected from seven locations at two different times in China were measured, and influences of rhizosphere soil on those minerals in plant were evaluated. The results showed that mean concentrations of eight minerals in plant samples decreased in the order: Ca > Mg > Na > K > Fe > Zn > Mn > Cu, and those in the soil samples followed the following order: Na > Fe > Ca > K > Mg > Mn > Zn > Cu. Mean concentrations of Ca, Na, Mg, and K in plants were higher than those in soils, while higher mean concentrations of the other four minerals were found in soils. It was found that there was a positive correlation of Mg, Na, and Cu concentrations in the plant with those in the soil respectively, but a negative correlation of Mn concentration in plant with that in the soil. Except Ca, K, and Mn, the other five minerals in plant were all directly affected by one or more chemical compositions of soil. The results also indicate that pH value and concentrations of total nitrogen, Mg, Mn, and Cu in soil had significant correlations with multimineral elements in plant. In a word, mineral elements uptake of S. divaricata can be changed by adjusting the soil fertility levels to meet the need of appropriate quality control of S. divaricata.