A new secondary metabolite from the marine-derived fungus Phomopsis lithocarpus FS508.

Based on the One Strain-Many Compounds (OSMAC) strategy, the secondary metabolites of Phomopsis lithocarpus FS508 were investigated. As a result, a new secondary metabolite, 4-methoxy-3-[4-(acetyloxy)-3-methyl-2-butenyl]benzoic acid (1) as well as eleven known compounds were isolated from the fermentation product of the strain FS508. Their structures were determined by NMR, IR, UV, and MS spectroscopic data analyses. All the isolated compounds were evaluated for cytotoxic and anti-inflammatory activities. Among them, compounds 3 and 9 displayed potent cytotoxicity against HepG-2 cell line, and compounds 2, 3 and 12 showed significant anti-inflammatory activities.

[Biosynthesis and metabolism regulation of terpenoids in Pogostemon cablin: a review].

The terpenoids in Pogostemon cablin have complex structures and abundant pharmacological effects. Patchouli alcohol(PA) and pogostone(PO) have a high medicinal value by virtue of anti-tumor, anti-inflammatory, antibacterial, antioxidant, and other biological activities. Due to the low content of terpenoid metabolites in P. cablin, the study of biosynthesis and metabolism regulation can provide a biosynthetic basis for obtaining high-content terpenoids. In this study, key enzyme genes in biosynthesis, transcription factors in metabolism regulation, spatio-temporal expression of terpene synthase were reviewed, aiming to provide a reference for the development, protection, and utilization of P. cablin resources.

[Effects of 5-azacytidine on DNA methylation and index components in patchouliol-type Pogostemon cablin].

This study aims to explore the relationship of DNA methylation with the contents of the index components as well as the growth and development of Pogostemon cablin. The demethylation reagent 5-azacytidine(5-azaC) was used to treat the tissue culture seedlings of patchouliol-type P. cablin. High performance liquid chromatography was employed to evaluate the changes of DNA methy-lation in P. cablin, and GC-MS to detect the contents of index components in P.cablin. The agronomic characters of P.cablin were measured using the common methods. The results showcased that DNA methylation of P.cablin was significantly reduced by 5-azaC in a concentration-dependent manner. Thirty days after treatment with 5-azaC at different concentrations, the content of patchouli alcohol changed slightly; compared with that in the control group, the content of pogostone in 50 µmol·L~(-1) and 100 µmol·L~(-1) 5-azaC groups was significantly up-regulated. The 100 µmol·L~(-1) 5-azaC group had the largest differences in contents of pogostone and patchouli alcohol compared with the control group, followed by the 50 µmol·L~(-1) 5-azaC group. Ninety days after disinhibition, the content of pogostone in the treatment group was significantly increased and the content of patchouli alcohol was significantly decreased. In addition, 5-azaC significantly inhibited the growth and development of P.cablin in a dose-dependent manner. These results indicate that DNA methylation regulates the biosynthesis of the index components in patchouliol-type P.cablin and proper demethylation can directly promote the synthesis of pogostone and indirectly affect the accumulation of patchouli alcohol.

In vitro induction and morphological characteristics of octoploid plants in Pogostemon cablin.

This paper describes an efficient colchicine-mediated technique for in vitro induction of octoploids in Pogostemon cablin and its confirmation by flow cytometry and chromosome numbers. The highest octoploid induction ratio was obtained by 0.05% colchicine treatment for 72 h. The chromosome number of octoploid seedlings was 2n = 8x = 128. Colchicine-induced tetraploids and octoploids planted in soil remained stable after 6 months. There were 31 lines of octoploid plants obtained. The leaf characteristics of P. cablin tetraploids and octoploids were compared. The larger leaves and stomata of transplants can be used to identify putative octoploids in P. cablin. Most octoploid lines exhibited higher patchoulic alcohol contents than the controls after 6 months of cultivation. Our results demonstrated that polyploidy induction can be beneficial in improving the medicinal value of P. cablin.

[Study on cytotoxic secondary metabolites of endophytic fungus Diaporthe longicolla A616 from Pogostemon cablin].

To study active secondary metabolites of endophytic fungus Diaporthe longicolla A616 isolated from Pogostemon cablin. Ten compounds were isolated from fermentation product of the strain 616 by silica gel, reverse phase silica gel, Sephadex-LH20, HPLC and so on. Their structures were identified as 1,3-diamino-1,3-dimethylurea(1),(7R,9R)-7-hydroxy-9-propyl-5-nonen-9-olide(2), Ergosta-5,7,22-trien-3ß-ol(3),(22E,24R)-ergosta-4,6,8(14)-22-tetraen-3-one(4),(22E,24R)-3ß,5α-dihydroxy-6ß-ergosta-7,22-diene(5), citreoisocoumarin(6), glycerol monolinoleate(7), 1-(2-hydroxyethoxy)ethyl(E)-octadec-9-enoate(8), cyclo-(L-Pro-L-Ala)(9), cyclo(L)-Pro-(L)-Val(10), respectively, based on extensive spectroscopic analysis and literature comparisons. Compounds 6-10 were isolated from the genus Diaporthe for the first time. All isolated compounds were evaluated for in vitro cytotoxic activities against SF-268, MCF-7, NCI-H460 and HepG-2 tumor cell lines. Compounds 4 and 5 showed potent growth inhibitory activities against the four cell lines with IC50 values of 5.3, 6.5, 12.2, 6.1µmol•L⁻¹ and 8.2, 5.2, 6.1, 9.4µmol•L⁻¹, respectively.

[Study on Chemical Constituents of Petroleum Ether Fraction from Rubus alceaefolius].

OBJECTIVE: To investigate the chemical constituents of Rubus alceaefolius. METHODS: Nine compounds were isolated and purified from the petroleum ether extract of 95% alcohol extract of Rubus alceaefolius by repeated column chromatography on silica, Sephadex LH-20 and structurally identified by spectral analysis. RESULTS: The compounds were identified as chrysophanol(1), physcion (2), ß-sitosterol(3), 3-oxotirucalla-7, 24-dien-21-oic acid(4), myricadiol(5), 19-α-hydroxy-3-acetyl-ursolic acid(6), N-benzoylphenylalaninyl-N-benzoylphenylalaninate(7), aurantiamide acetate(8) and euscaphic acid(9). CONCLUSION: Compounds land 4~8 are isolated from this plant for the first time, and compounds 4 - 8 are found in plants of Rubus genus for the first time.

[Cultural regionalization for Notopterygium incisum based on 3S technology platform. I. Evaluation for growth suitability for N. incisum based on ecological factors analysis by Maxent and ArcGIS model].

Growth suitability as assessment indicators for medicinal plants cultivation was proposed based on chemical quality determination and ecological factors analysis by Maxent and ArcGIS model. Notopterygium incisum, an endangered Chinese medicinal plant, was analyzed as a case, its potential distribution areas at different suitability grade and regionalization map were formulated based on growth suitability theory. The results showed that the most suitable habitats is Sichuan province, and more than 60% of the most suitable areawas located in the western Sichuan such as Aba and Ganzi prefectures for N. incisum. The results indicated that habitat altitude, average air temperature in September, and vegetation types were the dominant factors contributing to the grade of plant growth, precipitation and slope were the major factors contributing to notopterol accumulation in its underground parts, while isoimperatorin in its underground parts was negatively corelated with precipitation and slope of its habitat. However, slope as a factor influencing chemical components seemed to be a pseudo corelationship. Therefore, there were distinguishing differences between growth suitability and quality suitability for medicinal plants, which was helpful to further research and practice of cultivation regionalization, wild resource monitoring and large-scale cultivation of traditional Chinese medicine plants.

[Determination of anthraquinones from different medicinal parts of Berchemia lineata by HPLC].

OBJECTIVE: To develop an HPLC method for determination of emodin,chrysophanol and physcion from different medicinal parts of Berchemia lineata. METHODS: Samples were analyzed on Diamonsil ODS C18 (250 mm x 4. 6 mm,5 µm), with the mobile phase consisted of methanol-0. 20% phosphoric acid solution(74: 26). The flow rate was 1.0 mL/min,column temperature was set at 35 °C ,and detection UV wavelength was 254 nm. RESULTS: The linear range of emodin, chrysophanol and physcion was 0. 00201~ 0. 0804 µg,0. 0066~0. 264 µg and 0. 0124 ~0. 496 µg,with the average recovery was 100. 43% ,101. 29% and 98. 36% ,respectively. The content of total anthraquinones in root was higher than that in taten of Berchemia lineata. CONCLUSION: The method is simple,accurate and reliable for quality control of Berchemia lineata.

[Effects of agarwood formation induced by Fusarium sp. A2 on distribution and community composition of endophytic fungi in leaves of Aquilaria sinensis].

The aim of the paper is to observe the distribution of the endophytic fungi in leaves of Aquilaria sinensis by using permanent paraffin-cut section, optical microscope photography and histochemistry. Total DNA was extracted with modified CTAB method and rDNA ITS regions of plant and endophytic fungi were amplified with eukaryotic universal primers. The rDNA ITS amplicon was characterized by RFLP analysis, sequencing of rDNA ITS library and phylogenetic analyses using PAUP by maximum parsimony. Fusarium sp. A2 was used to induce the formation of resinous in A. sinensis trees. As a result, endophytic fungi mainly distributed in spongy and phloem in leaf. Endophytic fungi distributed in the phloem in agarwood-producing tree and had a relatively high abundance. Phoma sp. and Collectrotrichum sp. were the absolute advantage species in the leaf tissues of non-resinous and agarwood-producing tree, respectively. Collectrotrichum sp. was the only fungal species detected both in the two types of A. sinensis with different levels of abundance. The culture-independent molecular method can be used to identify fungal species directly and rapidly from the plant tissues. Endophytic fungal communities in non-resinous and agarwood-producing A. sinensis leaf tissues were quite different.

[Primary study on dormancy and release method for Berchemia lineata seed].

OBJECTIVE: To explore the mechanism of the seed dormancy of Berchemia lineatea and the method of breaking the dormancy. METHODS: Hundred-seed method, TTC and imbibition method was used to measure thousand seeds weight, seed viability and hard seed percentage, respectively. The germination inhibitor's biological characteristic was identified from the extract of every part in its fructification. Four treatments were compared to explore the best way to break physical and biological dormancy such as 98% sulfuric acid, hot water,grit friction and GA3. RESULTS: The thousand seeds weight was 10.82 g, the percentage of hard seed was up to 100%, viability was 83%, its water absorption speed and absorption rate were relatively low. Being treated with 98% sulfuric acid for 10 minutes, the seed improved its germination rate to 85%, and significantly improved its germination potentiality to 41%. Every part of the fructification contained germination inhibitors. The strongest inhibitory effect was found in seed testa and embryo. 500 and 1 000 mg/L GA3 significantly improved seed's germination potentiality to 48% and 43%. CONCLUSION: Berchemia lineata seed is hard to germinate because of its physical and biological dormancy, which is one reason for the resource reduction of Berchemia lineata.

[Pharmacognostical study on Berchemia floribunda].

OBJECTIVE: To study the pharmacognostical characteristics of stem and root of Berchemia floribunda for its further research and usage. METHODS: The plant was researched by macroscopic identification, microscopic identification and thin layer chromatography. RESULTS: The transverse section of B. floribunda root was eccentric. There were many fiber bundles in the secondary phloem and two different stone cells distributed in stem and root respectively. The results of TLC could identify the stem and root of B. floribunda. CONCLUSION: The microscopic characteristics of B. floribunda stem and root can be used as reference for its identification. Quercetin can be used as the characteristic component to identify the stem and root.

The correlation between bioactive components of Fallopia multiflora root and environmental factors.

Fallopia multiflora (Thunb.) Harald, a traditional Chinese medicinal plant, is used in treating dizziness. In this study, the samples of F. multiflora from ten different locations were collected, and five bioactive components (2, 3, 5, 4'-tetrahydroxystilbene-2-O-beta-D-glucoside, emodin, emodin-8-O-beta-D-glucoside, physcion and physcion-8-O-beta-D-glucoside) were quantified by high performance liquid chromatography. The correlations between 17 environmental factors and 5 bioactive components were analyzed. The results showed that the highest contents of bioactive components were in samples from Deqing, and the lowest in samples from Tianyang, which indicated that the quality of F. multiflora grown in Deqing was superior, while that grown in Tianyang was inferior. Emodin content was negatively correlated with the average temperature in January (p < 0.01) and the accumulated temperature (p < 0.01). Physician content was also negatively correlated with the average temperature in January (P < 0.01), the accumulated temperature (p < 0.05) and the organic matter (p < 0.05). However, emodin was positively correlated with the soil available K (p < 0.05) and Zn (p < 0.01). The results of stepwise regression showed that the accumulated temperature was the main factor influencing the contents of emodin and physcion. However, none of the environmental factors had significant correlation with 2, 3, 5, 4'-tetrahydroxystilbene-2-O-beta-D-glucoside, emodin-8- O-beta-D-glucoside and physcion-8-O-beta-D-glucoside. In conclusion, some environmental factors have significant influence on the content of dissociated anthraquinones, while some have no influence on that of combined anthraquinones.

[Study on determination and principal component analysis of inorganic elements in Polygonum multiflorum from different areas].

OBJECTIVE: To explore the character of inorganic elements in Polygonum multiflorum. METHOD: The contents of elements such as Al, B, Ba, Ca, Cu, Mn, Mg, Fe, Na, Ni, P, Se, Sr, Ti and Zn in nine P. multiflorum samples were determined by means of ICP-AEC. The results were used for the development of element distrubution diagram. The principal component analysis and one-way ANOVA of SPSS were applied for the study of characteristic elements in P. multiflorum. RESULT: The contents of Al, Ca, K, Mg, Sr,Ti in wild P. multiflorum were remarkable higher than those in cultured P. multiflorum, and there was no significant difference between cultured and wild in the other elements. Five principal components which accounted for over 90% of the total variance were extracted from the original data. The analysis results show that Al, B, Ba, Fe, Na, Ni, Ti, Ca and Sr may be the characteristic elements in P. multiflorum. The element distrubution diagram of the sample from Tianyang was remarkable different comparing with the others. CONCLUSION: The principal component analysis could be used in data processing in inorganic elements.

[Preliminary study on TLC fingerprint of radix polygoni multiflori from different areas].

OBJECTIVE: To establish thin-layer chromatography (TLC) fingerprint of Radix Polygoni Multiflori collected from different areas. METHODS: Quantitative analysis was carried out using TLC scanning method, two mixed solvents of petroleum ether (60-90 degrees C)-ethylacetate-formic acid (15:5:1) and chloroform-methanol-water [6.5: 2.25: 0.42, pH = 4.0 (HAc-NaAc buffer)] were employed as the mobile phase. RESULTS: The TLC chromatograms were constituted 14 characteristic common peaks. The plots based on the height of common peaks can be conveniently used to evaluate the quality of different area samples. CONCLUSION: The TLC fingerprints can be used to classify and identify Radix Polygoni Multiflori rapidly and effectively.