Four unreported aporphine alkaloids with antifungal activities from Artabotrys hexapetalus.

In this study, the extract from Artabotrys hexapetalus showed strong antifungal activity against phytopathogenic fungi in vitro. Four unreported aporphine alkaloids, hexapetalusine A-D (1-4), were isolated from stems and roots of Artabotrys hexapetalus (L.f.) Bhandari, along with six known aporphine alkaloids (5-10). Their chemical structures were elucidated by extensive spectroscopic analysis. The absolute configurations of 1-3 were determined using single-crystal X-ray diffractions and ECD calculations. Hexapetalusine A-C (1-3) were special amidic isomers. Additionally, all isolated compounds were evaluated for their antifungal activity against four phytopathogenic fungi in vitro. Hexapetalusine D (4) exhibited weak antifungal activity against Curvularia lunata. Liriodenine (5) displayed significant antifungal activity against Fusarium proliferatum and Fusarium oxysporum f. sp. vasinfectum, which is obviously better than positive control nystatin, suggesting that it had great potential to be developed into an effective and eco-friendly fungicide.

Chromosome level genome assembly of endangered medicinal plant Anisodus tanguticus.

Anisodus tanguticus is a medicinal herb that belongs to the Anisodus genus of the Solanaceae family. This endangered herb is mainly distributed in Qinghai-Tibet Plateau. In this study, we combined the Illumina short-read, Nanopore long-read and high-throughput chromosome conformation capture (Hi-C) sequencing technologies to de novo assemble the A. tanguticus genome. A high-quality chromosomal-level genome assembly was obtained with a genome size of 1.26 Gb and a contig N50 of 25.07 Mb. Of the draft genome sequences, 97.47% were anchored to 24 pseudochromosomes with a scaffold N50 of 51.28 Mb. In addition, 842.14 Mb of transposable elements occupying 66.70% of the genome assembly were identified and 44,252 protein-coding genes were predicted. The genome assembly of A. tanguticus will provide genetic repertoire to understand the adaptation strategy of Anisodus species in the plateau, which will further promote the conservation of endangered A. tanguticus resources.

Phytochemical Analysis of Nothapodytes tomentosa and Distribution and Content of Camptothecin and its Analogues in Four Plants.

Camptothecin (CPT) and its derivatives have attracted worldwide attention because of their notable anticancer activity. However, the growing demand for CPT in the global pharmaceutical industry has caused a severe shortage of CPT-producing plant resources. In this study, phytochemical analysis of Nothapodytes tomentosa results in the isolation and identification of CPT (13: ) and 16 analogues (1:  - 12, 14:  - 17: ), including a new (1: ) and five known (9, 10, 12, 15: , and 17: ) CPT analogues with an open E-ring. In view of the potential anticancer activity of CPT analogues with an open E-ring, the fragmentation pathways and mass spectra profiles of these six CPT analogues (1, 9, 10, 12, 15: , and 17: ) are investigated, providing a reference for the rapid detection of these compounds in other plants. Furthermore, based on the fragmentation patterns of CPT (13: ) and known analogues (2:  - 8, 11, 14, 16, 18:  - 26: ), the distribution and content of these compounds in different tissues of N. tomentosa, N. nimmoniana, Camptotheca acuminata, and Ophiorrhiza japonica are further studied. Our findings not only provide an alternative plant resource for further expanding the development and utilization of CPT and its analogues, but also lay a foundation for improving the utilization of known CPT-producing plant resources.

Products distribution during in situ and ex situ catalytic fast pyrolysis of Chinese herb residues.

Catalytic fast pyrolysis (CFP) for biomass treatment is a research hotspot but there is little information about the difference between the in situ and ex situ methods. In present work, the Ni-Fe/CaO-Al2O3 catalysts with different Ni/Fe ratios have been synthesized by coprecipitation method, and the product distribution about the Chinese herb residue (CHR) catalytic fast pyrolysis by in situ and ex situ methods in a quartz tube reactor system has been investigated. The results show that the CFP pyrolysis would upgrade the quality of bio-oil but decrease the yields, no matter in situ or ex situ CFP process. During the in situ CFP process, heteroatoms may be absorbed by the catalyst support and cannot be transferred to the bio-oil, but the results of ex situ CFP are the opposite. In addition, the ex situ CFP reaction significantly increases the content of aromatic hydrocarbons. As to the gas products' distribution, the effect of Fe in catalysts to promote CH4 formation is reflected in in situ CFP process, while the promotion effect of H2 generation for Ni added in catalyst is mainly reflected in ex situ CFP process. However, due to the high reaction temperature (800 °C), the adsorption of CO2 by CaO support is not particularly significant. The possible mechanism of CHR in CFP process has also been summarized for understanding the process of in situ and ex situ CFP, and this study may provide a choice or reference for CHR treatment.

Advances in chemistry and bioactivity of the genus Erythroxylum.

Erythroxylum P. Browne is the largest and most representative genus of Erythroxylaceae family. It contains approximately 230 species that are mainly distributed in tropical and subtropical regions. Some species in this genus, such as E. monogynum and E. coca, have been used as folk medicines in India or South America for a long history. It is well known that Erythroxylum plants are rich in tropane alkaloids, and the representative member cocaine shows remarkable activity in human central nervous system. However, many other types of active compounds have also been found in Erythroxylum along with the broadening and deepening of phytochemical research. To date, a total of 383 compounds from Erythroxylum have been reported, among which only 186 tropane alkaloids have been reviewed in 2010. In this review, we summarized all remained 197 compounds characterized from 53 Erythroxylum species from 1960 to 2021, which include diterpenes, triterpenes, alkaloids, flavonoids, and other derivates, providing a comprehensive overview of phytoconstituents profile of Erythroxylum plants. In addition, the biological activities of representative phytochemicals and crude extracts were also highlighted.

Co-pyrolysis of different torrefied Chinese herb residues and low-density polyethylene: Kinetic and products distribution.

In present work, the synergistic effects during co-pyrolysis of low-density polyethylene (LDPE) and torrefied Chinese herb residues (CHR) have been investigated by thermogravimetric analysis. The kinetic parameters of co-pyrolysis were calculated by Coats-Redfern method, and the difference values of experiment and theoretical were also investigated for gas and oil compounds. The results show that the extent of synergistic or inhibitory effects of co-pyrolysis was connection with the severity of CHR torrefaction, and the activation energy depend on the blend ratio of LDPE and CHRs. In addition, co-pyrolysis tends to generate more small molecule products and reduce oil yield, and increase the CO content but decreases CH4 in the gas product. The results also found that the liquid products have a significant interaction during the co-pyrolysis process, because the content of aliphatic hydrocarbons and alcohols in the blends pyrolysis oil has been greatly increased, and improving the quality of oil.

First report of Nigrospora sphaerica causing fruit dried-shrink disease in Akebia trifoliata from China.

Akebia trifoliata, a recently domesticated horticultural crop, produces delicious fruits containing multiple nutritional metabolites and has been widely used as medicinal herb in China. In June 2020, symptoms of dried-shrink disease were first observed on fruits of A. trifoliata grown in Zhangjiajie, China (110.2°E, 29.4°N) with an incidence about 10%. The infected fruits were shrunken, colored in dark brown, and withered to death (Figure S1A, B). The symptomatic fruits tissues (6 × 6 mm) were excised from three individual plants, surface-disinfested in 1% NaOCl for 30s and 70% ethanol solution for 45s, washed, dried, and plated on potato dextrose agar (PDA) containing 50 mg/L streptomycin sulfate in the dark, and incubated at 25℃ for 3 days. Subsequently, hyphal tips were transferred to PDA to obtain pure cultures. After 7 days, five pure cultures were obtained, including two identical to previously reported Colletotrichum gloeosporioides causing leaf anthracnose in A. trifoliata (Pan et al. 2020) and three unknown isolates (ZJJ-C1-1, ZJJ-C1-2, and ZJJ-C1-3). The mycelia of ZJJ-C1-1, ZJJ-C1-2 and ZJJ-C1-3 were white, and formed colonies of approximate 70 mm (diameter) in size at 25℃ after 7 days on potato sucrose agar (PSA) plates (Figure S1C). After 25 days, conidia were formed, solitary, globose, black, shiny, smooth, and 16-21 µm in size (average diameter = 18.22 ± 1.00 µm, n = 20) (Figure S1D). These morphological characteristics were similar to those of N. sphaerica previously reported (Li et al. 2018). To identify species of ZJJ-C1-1, ZJJ-C1-2 and ZJJ-C1-3, the internal transcribed spacer (ITS) region, ß-tubulin (TUB2), and the translation elongation factor 1-alpha (TEF1-α) were amplified using primer pairs including ITS1/ITS4 (Vilgalys and Hester 1990), Bt-2a/Bt-2b (Glass and Donaldson 1995), and EF1-728F/EF-2 (Zhou et al. 2015), respectively. Multiple sequence analyses showed no nucleotide difference was detected among genes tested except ITS that placed three isolates into two groups (Figure S2). BLAST analyses determined that ZJJ-C1-1, ZJJ-C1-2 and ZJJ-C1-3 had 99.73% to N. sphaerica strains LC2705 (KY019479), 100% to LC7294 (KY019397), and 99.79-100% to LC7294 (KX985932) or LC7294 (KX985932) based on sequences of TUB2 (MW252168, MW269660, MW269661), TEF-1α (MW252169, MW269662, MW269663), and ITS (MW250235, MW250236, MW192897), respectively. These indicated three isolates belong to the same species of N. sphaerica. Based on a combined dataset of ITS, TUB2 and TEF-1α sequences, a phylogenetic tree was constructed using Maximum likelihood method through IQ-TREE (Minh et al. 2020) and confirmed that three isolates were N. sphaerica (Figure S2). Further, pathogenicity tests were performed. Briefly, healthy unwounded fruits were surface-disinfected in 0.1% NaOCl for 30s, washed, dried and needling-wounded. Then, three fruits were inoculated with 10 µl of conidial suspension (1 × 106 conidia/ml) derived from three individual isolates, with another three fruits sprayed with 10 µl sterilized water as control. The treated fruits were incubated at 25℃ in 90% humidity. After 15 days, all the three fruits inoculated with conidia displayed typical dried-shrink symptoms as those observed in the farm field (Figure S1E). The decayed tissues with mycelium and spores could be observed on the skin or vertical split of the infected fruits after 15 days' inoculation (Figure S1F-H). Comparably, in the three control fruits, there were no dried-shrink-related symptoms displayed. The experiment was repeated twice. The re-isolated pathogens were identical to N. sphaerica determined by sequencing the ITS, TUB2 and TEF-1α. Previous reports showed N. sphaerica could cause postharvest rot disease in kiwifruits (Li et al. 2018). To our knowledge, this is the first report of N. sphaerica causing fruits dried-shrink disease in A. trifoliata in China.

Characterization of inthomycin biosynthetic gene cluster revealing new insights into carboxamide formation.

Inthomycins are polyketide antibiotics which contain a terminal carboxamide group and a triene chain. Inthomycin B (1) and its two new analogues 2 and 3 were isolated from the crude extract of Streptomyces pactum L8. Identification of the gene cluster for inthomycin biosynthesis as well as the 15N-labeled glycine incorporation into inthomycins are described. Combined with the gene deletion of the rare P450 domain in the NRPS module, a formation mechanism of carboxamide moiety in inthomycins was proposed via an oxidative release of the assembly chain assisted by the P450 domain.

The complete chloroplast genome of Akebia trifoliata (Lardizabalaceae), a traditional herb in China.

Akebia trifoliata, commonly known as 'Bayuezha' in China, has been widely used as traditional Chinese medicinal herbs with a long history. In the present study, the complete chloroplast genome of A. trifoliata was sequenced using Illumina high-throughput sequencing approach. The length of the complete chloroplast genome is 157,952 bp with 38.7% GC content. It contains 131 genes, including 86 protein-coding genes, 37 tRNA genes, and 8 rRNA genes. Phylogenetic analysis indicated that A. trifoliata was closely related to another Lardizabalaceae species, Akebia quinata, which further confirms traditional species classification.

Functional genomics analysis reveals two novel genes required for littorine biosynthesis.

Some medicinal plants of the Solanaceae produce pharmaceutical tropane alkaloids (TAs), such as hyoscyamine and scopolamine. Littorine is a key biosynthetic intermediate in the hyoscyamine and scopolamine biosynthetic pathways. However, the mechanism underlying littorine formation from the precursors phenyllactate and tropine is not completely understood. Here, we report the elucidation of littorine biosynthesis through a functional genomics approach and functional identification of two novel biosynthesis genes that encode phenyllactate UDP-glycosyltransferase (UGT1) and littorine synthase (LS). UGT1 and LS are highly and specifically expressed in Atropa belladonna secondary roots. Suppression of either UGT1 or LS disrupted the biosynthesis of littorine and its TA derivatives (hyoscyamine and scopolamine). Purified His-tagged UGT1 catalysed phenyllactate glycosylation to form phenyllactylglucose. UGT1 and LS co-expression in tobacco leaves led to littorine synthesis if tropine and phenyllactate were added. This identification of UGT1 and LS provides the missing link in littorine biosynthesis. The results pave the way for producing hyoscyamine and scopolamine for medical use by metabolic engineering or synthetic biology.

Cangumycins A-F, six new angucyclinone analogues with immunosuppressive activity from Streptomyces.

Cangumycins A-F (1-6), six new angucyclinone analogues, together with two known ones (7 and 8), were isolated from the fermentation broth of a soil-derived Streptomyces sp. KIB-M10. Structures of these compounds were elucidated via a joint use of spectroscopic analyses and single-crystal X-ray diffractions. Among them, cangumycins E (5) and F (6) share a C-ring cleaved backbone, and cangumycins B (2) and E (5) exhibit potent immunosuppressive activity (IC50 8.1 and 2.7 µmol·L-1, respectively) against human T cell proliferation at a non-cytotoxic concentration.

A new actinomycin Z analogue with an additional oxygen bridge between chromophore and ß-depsipentapeptide from Streptomyces sp. KIB-H714.

Actinomycin Z6 (1), a new member of the actinomycin family, along with three congeners of the Z-type (Z1, Z3, Z5) actinomycins, are produced from Streptomyces sp. KIB-H714. Their structures were authenticated by comprehensive spectroscopic data interpretation. Different from all the reported Z-type actinomycins, the ß-ring of the new compound actinomycin Z6 includes an additional ring linked between the actinoyl chromophore and ß-peptidolactone. In Z3 and Z5, the L-threonine in ß-depsipeptide is replaced by the unusual 4-chlorothreonine, an amino acid rarely found in actinomycin family. All isolates were evaluated for cytotoxicity against five human tumor cell lines and for inhibitory activity against Candida albicans and Staphylococcus aureus.

New phloroglucinol derivatives with protein tyrosine phosphatase 1B (PTP1B) inhibitory activities from Syzygium austroyunnanense.

Five new phloroglucinol derivatives (1-5) together with one known analogue (6) were isolated from the leaves of Syzygium austroyunnanense which is an edible folk medicine used for the treatment of diabetes. The new structures were elucidated as austroyunones A - E (1-5) by means of the extensive spectroscopic analyses including high-resolution electrospray ionization mass spectrometry (HRESIMS) and 1D and 2D nuclear magnetic resonance (NMR). Compounds 4-6 showed obvious protein tyrosine phosphatase 1B (PTP1B) inhibitory activity. This discovery of new phloroglucinols and their bioactivities provided a scientific basis for the application of S. austroyunnanense as an edible and medicinal plant.

Cytotoxicity-Guided Isolation of Two New Phenolic Derivatives from Dryopteris fragrans (L.) Schott.

Dryopteris fragrans is a valuable medicinal plant resource with extensive biological activities including anti-cancer, anti-oxidation, and anti-inflammation activities. This work aims to study further the cytotoxic constituents from Dryopteris fragrans. In this work, two new phenolic derivatives known as dryofragone (1) and dryofracoumarin B (2) with six known compounds (3⁻8) were isolated from the petroleum ether fraction of the methanol extract of the aerial parts of Dryopteris fragrans (L.) Schott by two round cytotoxicity-guided tracking with the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay and cell counting kit-8 (CCK-8) assay. Their structures were elucidated by the extensive spectroscopic analysis (¹H-NMR, 13C-NMR, and two dimensions NMR), chemical derivatization, and comparison with data reported in the literature. All the isolates were evaluated for their cytotoxicity against nine cancer cell lines as well as their in vitro immunomodulatory activity. The results showed that compounds have a modest cytotoxicity toward human HeLa cell line with IC50 value below 30 µM and compounds 4 and 5 may modulate immunity to affect the growth of tumor cells.

Determination of garlic phenolic compounds using supercritical fluid extraction coupled to supercritical fluid chromatography/tandem mass spectrometry.

A rapid, simple and environmentally friendly supercritical fluid extraction and supercritical fluid chromatography coupled to mass spectrometry (SFE-SFC-MS/MS) method has been developed for the analysis of nine phenolic compounds in garlic. Through the optimization of the SFE parameters using response surface methodology (RSM), 15 phenolic compounds were successfully extracted at 50 °C in 9 min with the addition of 30% methanol using a method that was automatic, efficient, green and prevented oxidation. Moreover, the operating conditions of the SFC-MS/MS were also optimized for the analysis of the SFE extracts. 15 phenolic compounds in the garlic showed good separation performance on a Shim-pack UC-X Diol column, and 0.1 mM oxalic acid and 1 mM ammonium formate in methanol were chosen as the most suitable mobile phase components. Furthermore, a calibration curve was developed using a matrix-matched standard, and 9 phenolic compounds showed good linearity (R2 > 0.99). The limits of detection of the 9 phenolic compounds varied from 0.6 to 12 ng/g, and the quantifications ranged from 2 to 40 ng/g. The validation data also showed acceptable recoveries for 9 phenolic compounds, and the relative standard deviations (RSDs) for the overall precision were lower than 10%. The developed method has been utilized to analyze the phenolic compounds in garlic from different geographical areas of China (Cangshan, Jinxiang, Pizhou, Dali), which suggests that this off-line SFE-SFC-MS/MS method could be adapted to the quantitative analysis of phenolic compounds in garlic.

Bioactive acylphloroglucinols with adamantyl skeleton from Hypericum sampsonii.

Hyperisampsins A-D (1-4), with tetracyclo[6.3.1.1(3,10).0(3,7)]tridecane skeletons and seven biogenetically related congeners (5-11), were isolated from Hypericum sampsonii. Their structures were elucidated by comprehensive spectroscopic techniques. The absolute configuration of 1 was established by ECD calculations, and those of 5 and 9 were confirmed by single X-ray crystallographic analyses. Hyperisampsins A and D showed potent anti-HIV activities with EC50 of 2.97 and 0.97 µM and selectivity index of 4.80 and 7.70, respectively.

19-oxygenated ent-kaurane diterpenoids from Isodon pharicus.

Eight new 19-oxygenated ENT-kaurane diterpenoids were isolated from the aerial parts of Isodon pharicus. Their structures were determined by means of extensive spectroscopic techniques including interpretation of 1D and 2D NMR spectra. Selected compounds were evaluated for their cytotoxicity against NB4, A549, PC-3, MCF-7, and SH-SY5Y cell lines.

Three new arylnaphthalene lignans from Schisandra propinqua var. sinensis.

Three new arylnaphthalene lignans, named sinensisins A-C (1-3), together with three known compounds, were isolated from the aerial parts of Schisandra propinqua var. sinensis. Their structures were established by spectroscopic methods, and compound 1 exhibited weak anti-HIV-1 activity with an TI value of 6.7.

Schisanartane nortriterpenoids with diverse post-modifications from Schisandra propinqua.

Twenty-one highly oxygenated nortriterpenoids with a schisanartane skeleton were isolated from the stems of Schisandra propinqua var. propinqua. These nortriterpenoids featured a polycyclic framework composed of 7/8/5 consecutive carbocycles, which were organized by similar 5/5/7/5/7/5 rings A-F and varied oxygen-containing rings G and H. A biosynthetic classification of these compounds is proposed on the basis of their diverse post-modifications.

Bisnortriterpenoids possessing an 18-Nor-schiartane skeleton from Schisandra propinqua var. propinqua.

Six C (28) triterpenoids with an 18-nor-schiartane framework were isolated from the aerial parts of SCHISANDRA PROPINQUA var. PROPINQUA. The new compounds, propindilactones P-S ( 1- 4), were structurally characterized by extensive spectroscopic methods. New compounds 2 and 3, together with known wuweizidilactones B and H ( 5- 6), were evaluated for their anti-HBV activity.