Plant Virome Analysis by the Deep Sequencing of Small RNAs of Fritillaria thunbergii var. chekiangensis and the Rapid Identification of Viruses.

Thunberg fritillary (Fritillaria thunbergii), a perennial used in traditional Chinese herbal medicine, is a members of the family Liliaceae. The degeneration of germplasm is a severe problem in the production of Fritillaria thunbergii var. chekiangensis. However, no information about viral infections of F. thunbergii var. chekiangensis has been reported. In this study, we sequenced the small RNAs of F. thunbergii var. chekiangensis from leaves and bulbs, and viruses were identified using a phylogenetic analysis and BLAST search for sequence. In addition, multiplex reverse transcriptase-polymerase chain reaction (RT-PCR) was used to rapidly detect viruses in this variety. Our study first reported that five viruses infected F. thunbergii var. chekiangensis. Among them, fritillary virus Y (FVY), lily mottle virus (LMoV), Thunberg fritillary mosaic virus (TFMV), and hop yellow virus (HYV) had been reported in F. thunbergii, while apple stem grooving virus was first reported in the genus Fritillaria. A multiplex RT-PCR method was developed to rapidly test the four viruses FVY, LMoV, TFMV, and HYV in F. thunbergii var. chekiangensis. Our results provide a better understanding of the infection of F. thunbergii var. chekiangensis by viruses and a basic reference for the better design of suitable control measures.

Tandem mass spectrometric analysis and density functional theory calculations on the fragmentation behavior of two tetradecanoylingenol regioisomers from Euphorbia wallichii.

RATIONALE: Two structurally similar bioactive regioisomers, 3-O-tetradecanoylingenol and 20-O-tetradecanoylingenol, from Euphorbia wallichii presented quite different fragmentation behaviors. Revealing the related fragmentation pathways may improve the efficiency of characterization and identification of such type of compounds. METHODS: The two regioisomers were subjected to collision-induced dissociation (CID) on Finnigan LCQ(DECA) and LTQ Orbitrap XL instruments. Based on the CID results, the possible fragmentation pathways were proposed and investigated with density functional theory (DFT) calculations. RESULTS: Elimination of C(14)H(28)O(2) (tetradecanoic acid) for 3-O-tetradecanoylingenol and the sequential losses of C(4)H(8) (butylene) for 20-O-tetradecanoylingenol were observed in ESI-MS/MS, witnessed also by HR-ESI-MS/MS. The fragmentation pathways were proposed and verified by calculating the activation energy of their transition states by DFT calculations. CONCLUSIONS: Based on the observations, fragmentation pathways for the two regioisomers were proposed and verified by calculating the energy of the reactants, products and the corresponding transition states using DFT. This report should have value in rapid identification of the derivatives of ingenol and other regioisomers in natural products.

[Chemical constituents contained in seeds of Notopterygium franchetii].

OBJECTIVE: To study the chemical constituents from the seeds of Notopterygium franchetii. METHOD: Ethanol extracts of seeds N. franchetii were separated and purified by such methods as normal and reversed phase column chromatographies and thin-layer chromatography and structurally elucidated by MS and NMR evidences. RESULT: Twenty nine compounds were separated, they were isoimperatorin (1), [3-sitosterol (2), phellopterin (3), bergapten (4), N-tetra, hexa, octacosanoylanthranilic acid (5-7), daucosterol (8), oxypeucedanin hydrate (9), umbelliferone (10), demethylfuropinnarin (11), (2S, 3S, 4R, 8E)-2-[(2'R)- 2'-hydroxydoco, trico, tetraco, entaco, hexaco sanosylamino] -octadecene-1, 3, 4-triol (12-16), (-)-oxypeucedanin (17), diosmetin (18), bergaptol-O-beta-D-glucopyranoside (19), nodakenin (20), 1'-O-beta-D-glucopyranosyl-(2R, 3S)-3-hydroxynodakenetin (21), uracil (22), decuroside V (23), 8-O-beta-D-glucopyranosyl-5-hydroxypsoralen (24), 8-O-beta-D-glucopyranosyl-5-methoxylpsoralen (25), diosmin (26), alaschanioside C (27), kynurenic acid (28) and mannitol (29). CONCLUSION: All of these compounds were separated from the seeds of N. franchetii for the first time. Of them, 18, 22, 26 and 29 were firstly obtained from genus Notopterygium.

Discrimination of the seeds of Notopterygium incisum and Notopterygium franchetii by validated HPLC-DAD-ESI-MS method and principal component analysis.

A validated HPLC-DAD-ESI-MS method has been developed to simultaneously quantify 12 bioactive compounds in the seeds of Notopterygium incisum Ting ex H.T. Chang and Notopterygium franchetii H. de Boiss whose rhizomes and roots are widely used as traditional Chinese medicine. This method was validated to be sensitive, precise and accurate and was applied to evaluate the difference in the chemical profiles and contents of these analytes in 37 batches of N. incisum and 31 batches of N. franchetii samples collected from different locations. Principal component analysis showed that the two species were separated into two groups obviously. This study established a validated method for identification of the authenticity of the seeds of N. incisum and N. franchetii and supplied effective guidance for artificial cultivation.

Three new flavone C-glycosides from the aerial parts of Paraquilegia microphylla.

Three new flavone C-glycosides, paraquinins A-C, were isolated from the aerial parts of Paraquilegia microphylla (Royle) Dromm. et Hutch, a Tibetan medicine distributed in the Qinghai-Tibet plateau. On the basis of 1D and 2D NMR evidence, their structures were elucidated as acacetin-6-C-ß-D-glucopyranosyl-(1 → 2)-ß-D-glucopyranoside (1), acacetin-6-C-α-L-rhamnopyranosyl-(1 → 2)-ß-D-glucopyranosyl-(1 → 2)-ß-D-glucopyranoside (2), and acacetin-6-C-α-L-rhamnopyranosyl-(1 → 2)-(6'''-O-E-feruloyl)-ß-D-glucopyranosyl-(1 → 2)-ß-D-glucopyranoside (3).

Two new triterpene saponins from the aerial parts of Paraquilegia microphylla.

Two new triterpene saponins, paraquinosides A (1) and B (2) were isolated from the aerial parts of Paraquilegia microphylla (Royle) Dromm. et Hutch, a Tibetan ethnic medicine distributed in the Qinghai-Tibet Plateau. On the basis of 1D and 2D NMR evidence, their structure was elucidated as 3-O-alpha-L-rhamnopyranosyl (1-->2)-beta-D-glucopyranosyl-15-dehydroxyl-16-O-methyl-24, 25-deoxy-26-hydroxylshengmanol-26-O-beta-D-glucopyranoside (1) and 3-O-alpha-L-Rhamnopyranosyl (1-->2)-[beta-D-glucopyranosyl(1-->3)]-beta-D-glucopyranosyl-15-dehydroxyl-16-O-methyl-24, 25-deoxy-26-hydroxylshengmanol-26-O-beta-D- glucopyranoside (2), respectively.

Quantitative analysis of chemical constituents in different commercial parts of Notopterygium incisum by HPLC-DAD-MS.

ETHNOPHARMACOLOGICAL RELEVANCE: Notopterygium incisum used as an antirheumatic and analgesic medicine for the treatment of rheumatism, headache, cold, etc., is not only a traditional Qiang medicine, but also a traditional Tibetan remedy for thousands of years. AIM OF THE STUDY: In order to use this herbal medicine in a more effective way, in the present investigation, we analyzed the main chemical constituents of different commercial parts as well as fibrous roots of Notopterygium incisum. MATERIALS AND METHODS: All crude Notopterygium incisum samples used in this study were collected in Sichuan province of China. We used HPLC-DAD-MS to analyze quantitatively the main chemical constituents of different commercial parts as well as fibrous roots of Notopterygium incisum. RESULTS: According to different commercial parts, 10 Notopterygium incisum samples collected from two main production areas have been investigated, and four main constituents have been quantitatively analyzed. CONCLUSION: The quality of rhizomes is better than that of main roots. Unexpectedly, the content of two main constituents, notopterol and isoimperaorin, in fibrous roots is higher than any other parts of Notopterygium incisum. This result implied that the fibrous root may also have value, and we should study it to explore its potential.