The Liquid Fermentation Process for Mycelia of Poria cocos (Agaricomycetes) by Single-Factor Experimentation and Response Surface Methodology.

Liquid fermentation could yield substantial mycelia mass and valuable secondary metabolites in large-scale production within a short, fermented duration. The liquid fermented process of mycelia of Poria cocos was optimized using a combination of single-factor experimentation and response surface methodology (RSM) to obtain more extract of P. cocos. The optimal conditions were determined as follows: The carbon source concentration at 1%, the nitrogen source concentration at 1%, the inoculum volume at 7% and a culture time of 9 d. Under these conditions, the ethyl acetate extract mass of P. cocos mycelia reached 0.0577 ± 0.0041 mg. There were significant interactions between nitrogen source concentration and cultivation time. The predicted values by the mathematical model based on the response surface analysis showed a close agreement with experimental data.

Chemical Constituents from Mycelia of Lepista sordida (Agaricomycetes) and Their ABTS Radical Scavenging Activity.

Lepista sordida is an edible mushroom possessing high nutritional value and high medicinal value. The artificial cultivation technology of L. sordida made a breakthrough and has been popularized in Yunnan, Guizhou, Sichuan province with good economic benefits. The secondary metabolites were investigated from ethyl EtOAc (acetate extract) of solid cultures of L. sordida. Silica gel column chromatography, semi-preparation HPLC, recrystallization, and medium pressure column chromatography were applied to obtain 15 compounds. Nine compounds were first isolated from genus Lepista and 11 compounds were first isolated from species L. sordida. Moreover, compounds 13 and 14 exhibited strong scavenging activity of ABTS.

Characterization of Secondary Metabolites from Mycelial Cultures of Black Morel Mushroom Morchella importuna (Ascomycota).

Species of the genus Morchella are highly prized worldwide for their excellent flavor and high medicinal value. In recent years, artificial cultivations of medicinal fungi with many advantages have elicited great interest as a promising alternative to produce certain valuable metabolites. Therefore, the secondary metabolites of fermented M. importuna belonging to the black morel clade isolated from China were investigated. The strain was cultured in a fermentation tank in PDB liquid medium by two-step method. The mycelia and fermentation broth were extracted by ethyl acetate. The secondary metabolites were separated and purified by repeated silica gel column chromatography. Structures of compounds were determined by NMR data and references. One new natural compound (1) and six known compounds (2-7) were obtained. Compounds 1, 2, 4, and 5 were first isolated from genus Morchella and compounds 3, 6, and 7 are first isolated from species M. importuna.

Investigation of Chemical Compounds and DPPH Radical Scavenging Activity of Oudemansiella raphanipes (Agaricomycetes) Based on Fermentation.

Oudemansiella raphanipes, an edible medicinal mushroom, is very popular in the market. With the development of technology, liquid fermentation is becoming more and more popular in edible fungi cultivation. Strain O. raphanipes was cultured by submerged fermentation and secondary metabolites were investigated simultaneously. Six chemical constituents including two unsaturated fatty acids (1-2), one ergosterol (3), one phenol (4), and two benzamides (5-6) were acquired from ethyl acetate extract. Their structures were mainly elucidated by NMR spectral data and known products. All compounds were isolated from O. raphanipes for the first time. DPPH radical scavenging activity was evaluated. Orcinol (4) exhibited remarkable antioxidant activity; its free radical scavenging rate can reach up to 80% of 20 mg/mL. 2-pyruvoybenzamide (5) and 2-(2-hydroxypropanamido) benzamide (6) showed less active antioxidant capacity.

Comparative phosphoproteome analysis upon ethylene and abscisic acid treatment in Glycine max leaves.

Abscisic acid (ABA) and ethylene play key roles in growth and development of plants. Several attempts have been made to investigate the ABA and ethylene-induced signaling in plants, however, the involvement of phosphorylation and dephosphorylation in fine-tuning of the induced response has not been investigated much. Here, a phosphoproteomic analysis was carried out to identify the phosphoproteins in response to ABA, ethylene (ET) and combined ABA + ET treatments in soybean leaves. Phosphoproteome analysis led to the identification of 802 phosphopeptides, representing 422 unique protein groups. A comparative analysis led to the identification of 40 phosphosites that significantly changed in response to given hormone treatments. Functional annotation of the identified phosphoproteins showed that these were majorly involved in nucleic acid binding, signaling, transport and stress response. Localization prediction showed that 67% of the identified phosphoproteins were nuclear, indicating their potential involvement in gene regulation. Taken together, these results provide an overview of the ABA, ET and combined ABA + ET signaling in soybean leaves at phosphoproteome level.

Harvesting more grain zinc of wheat for human health.

Increasing grain zinc (Zn) concentration of cereals for minimizing Zn malnutrition in two billion people represents an important global humanitarian challenge. Grain Zn in field-grown wheat at the global scale ranges from 20.4 to 30.5 mg kg-1, showing a solid gap to the biofortification target for human health (40 mg kg-1). Through a group of field experiments, we found that the low grain Zn was not closely linked to historical replacements of varieties during the Green Revolution, but greatly aggravated by phosphorus (P) overuse or insufficient nitrogen (N) application. We also conducted a total of 320-pair plots field experiments and found an average increase of 10.5 mg kg-1 by foliar Zn application. We conclude that an integrated strategy, including not only Zn-responsive genotypes, but of a similar importance, Zn application and field N and P management, are required to harvest more grain Zn and meanwhile ensure better yield in wheat-dominant areas.

Inhibitory effect of bufalin on retinoblastoma cells (HXO-RB44) via the independent mitochondrial and death receptor pathway.

Cinobufacini (Huachansu) is a Chinese medicine prepared from the skin of Bufo bufo gargarizans Cantor (Bufonidae), and has long been used in traditional Chinese medicine. In the present study, the anti-retinoblastoma constituent bufalin obtained from Cinobufacini was investigated. Treatment of human retinoblastoma (HXO-RB44) cells with bufalin induced apoptosis which was accompanied by a decrease in mitochondrial membrane potential, activation of caspase-9, caspase-8 and caspase-3, as well as changes in the expression of cytochrome C. Bufalin induced the cleavage of caspase-3 and apoptosis, and it was inhibited by both Z-LETD-FMK and Z-IETD-FMK treatment. Taken together, these results demonstrate that bufalin-induced apoptosis in human retinoblastoma (HXO-RB44) cells involved both intrinsic and extrinsic pathways.

Isolation of active compounds from methanol extracts of Toddalia asiatica against Ichthyophthirius multifiliis in goldfish (Carassius auratus).

The parasitic ciliate Ichthyophthirius multifiliis infests all species of freshwater fish and can cause severe economic losses in fish breeding. The present study aims to evaluate the antiparasitic activity of the active components from Toddalia asiatica against I. multifiliis. Bioassay-guided fractionation and isolation of compounds with antiparasitic activity were performed on the methanol extract of T. asiatica yielding two bioactive compounds: chelerythrine and chloroxylonine identified by comparing spectral data (NMR and ESI-MS) with literature values. Results from in vitro antiparasitic assays revealed that chelerythrine and chloroxylonine could be 100% effective against I. multifiliis at the concentration of 1.2 mg L(-1) and 3.5 mg L(-1), with the median effective concentration (EC50) values of 0.55 mg L(-1) and 1.90 mg L(-1) respectively. In vivo experiments demonstrated that fish treated with chelerythrine and chloroxylonine at the concentrations of 1.8 and 8.0 mg L(-1) carried significantly fewer parasites than the control (P<0.05). The acute toxicity (LC50) of chelerythrine for goldfish was 3.3 mg L(-1).

Daidzein induced apoptosis via down-regulation of Bcl-2/Bax and triggering of the mitochondrial pathway in BGC-823 cells.

Daidzein belongs to the group of isoflavones, found in a wide variety of plant-derived foods, especially in soybeans and soy-based foods. In this study, the effect of daidzein on human gastric carcinoma cells (BGC-823) and its mechanism were investigated. MTT assay was applied in the detection of the inhibitory effects of daidzein on cell proliferation. Hoechst-propidium iodide staining and flow cytometry were used to examine the apoptosis as well as the mitochondrial transmembrane potential. Western blotting was performed to detect the expression of apoptosis-associated proteins: cleaved PARP, cleaved caspase-9, cleaved caspase-3, Bcl-2, and Bax. Daidzein significantly inhibited the growth and proliferation of human gastric carcinoma cells (BGC-823) in a concentration- and time-dependent manner. Furthermore, it was found that an insult of daidzein to BGC-823 cells caused them to die by disruption of mitochondrial transmembrane potential, demonstrated not only by staining dead cells for phosphatidylserine but also by the up-regulation (cleaved PARP, cleaved caspase-9, cleaved caspase-3, Bax) and down-regulation (Bcl-2) of proteins associated with apoptosis and survival; whereas, the pan-caspase inhibitor z-VAD-fmk could partially rescue cells against damage of daidzein. Taken together, the results of this study demonstrate that daidzein significantly induces apoptosis via a mitochondrial pathway. Specifically, daidzein induced a change in the Bax/Bcl-2 ratios and activation of caspases-3 and -9 and the cleavage of PARP. Therefore, daidzein has the potential for use as a therapeutic agent for the treatment of gastric carcinoma.

Multihydroxylation of ursolic acid by Pestalotiopsis microspora isolated from the medicinal plant Huperzia serrata.

The structural modification of ursolic acid by an endophytic fungus Pestalotiopsis microspora, isolated from medicinal plant Huperzia serrata was reported for the first time. The structure diversity was very important for the SAR study of ursolic acid and its derivatives. Incubation of ursolic acid 1 with P. microspora afforded four metabolites: 3-oxo-15α, 30-dihydroxy-urs-12-en-28-oic acid (2), 3ß, 15α-dihydroxy-urs-12-en-28-oic acid (3), 3ß, 15α, 30- trihydroxy-urs-12-en-28-oic acid (4) and 3,4-seco-ursan-4,30-dihydroxy-12-en-3,28-dioic acid (5). All products were new compounds and their structures elucidation was mainly based on the spectroscopic data.