Effect of Different Carbon Sources on Antioxidant Properties of Exopolysaccharides Produced by Scleroderma areolatum (Agaricomycetes).

Five kinds of exopolysaccharides (EPS) were obtained by fermentation of Scleroderma areolatum Ehrenb. with sucrose, glucose, maltose, lactose, and fructose as carbon sources. Antioxidant abilities of the obtained EPSs were evaluated by inhibiting AAPH, HO·, and glutathione (GS·) induced oxidation of DNA and quenching 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonate) cationic radical (ABTS· and galvinoxyl radicals. The effects of carbon sources on the antioxidant properties of EPSs could be examined. The results showed that five EPSs can effectively inhibit radicals induced oxidation of DNA, and the thiobarbituric acid reactive substances (TBARS) percentages were 44.7%-80.8%, 52.3%-77.5%, and 44.7%-73.3% in inhibiting AAPH, HO·, and GS· induced oxidation of DNA, respectively. All five EPSs could scavenge ABTS· and galvinoxyh, and exhibit superior activity in scavenging free radicals. Antioxidant abilities of EPS with fructose as carbon source were highest among five EPS.

Phylogenomic analyses based on the plastid genome and concatenated nrDNA sequence data reveal cytonuclear discordance in genus Atractylodes (Asteraceae: Carduoideae).

Atractylodes species are widely distributed across East Asia and are cultivated as medicinal herbs in China, Japan, and Korea. Their unclear morphological characteristics and low levels of genetic divergence obscure the taxonomic relationships among these species. In this study, 24 plant samples were collected representing five species of Atractylodes located in China; of these, 23 belonged to members of the A. lancea complex. High-throughput sequencing was used to obtain the concatenated nrDNA sequences (18S-ITS1-5.8S-ITS2-28S) and plastid genomes. The concatenated nrDNA sequence lengths for all the Atractylodes species were 5,849 bp, and the GC content was 55%. The lengths of the whole plastid genome sequences ranged from 152,138 bp (A. chinensis) to 153,268 bp (A. lancea), while their insertion/deletion sites were mainly distributed in the intergenic regions. Furthermore, 33, 34, 36, 31, and 32 tandem repeat sequences, as well as 30, 30, 29, 30, and 30 SSR loci, were detected in A. chinensis, A. koreana, A. lancea, A. japonica, and A. macrocephala, respectively. In addition to these findings, a considerable number of heteroplasmic variations were detected in the plastid genomes, implying a complicated phylogenetic history for Atractylodes. The results of the phylogenetic analysis involving concatenated nrDNA sequences showed that A. lancea and A. japonica formed two separate clades, with A. chinensis and A. koreana constituting their sister clade, while A. lancea, A. koreana, A. chinensis, and A. japonica were found based on plastid datasets to represent a mixed clade on the phylogenetic tree. Phylogenetic network analysis suggested that A. lancea may have hybridized with the common ancestor of A. chinensis and A. japonica, while ABBA-BABA tests of SNPs in the plastid genomes showed that A. chinensis was more closely related to A. japonica than to A. lancea. This study reveals the extensive discordance and complexity of the relationships across the members of the A. lancea complex (A. lancea, A. chinensis, A. koreana, and A. japonica) according to cytonuclear genomic data; this may be caused by interspecific hybridization or gene introgression.

A chromosome-level genome assembly for Dracaena cochinchinensis reveals the molecular basis of its longevity and formation of dragon's blood.

Dracaena, a remarkably long-lived and slowly maturing species of plant, is world famous for its ability to produce dragon's blood, a precious traditional medicine used by different cultures since ancient times. However, there is no detailed and high-quality genome available for this species at present; thus, the molecular mechanisms that underlie its important traits are largely unknown. These factors seriously limit the protection and regeneration of this rare and endangered plant resource. Here, we sequenced and assembled the genome of Dracaena cochinchinensis at the chromosome level. The D. cochinchinensis genome covers 1.21 Gb with a scaffold N50 of 50.06 Mb and encodes 31 619 predicted protein-coding genes. Analysis showed that D. cochinchinensis has undergone two whole-genome duplications and two bursts of long terminal repeat insertions. The expansion of two gene classes, cis-zeatin O-glucosyltransferase and small auxin upregulated RNA, were found to account for its longevity and slow growth. Two transcription factors (bHLH and MYB) were found to be core regulators of the flavonoid biosynthesis pathway, and reactive oxygen species were identified as the specific signaling molecules responsible for the injury-induced formation of dragon's blood. Our study provides high-quality genomic information relating to D. cochinchinensis and significant insight into the molecular mechanisms responsible for its longevity and formation of dragon's blood. These findings will facilitate resource protection and sustainable utilization of Dracaena.

Identification of the original plants of cultivated Bupleuri Radix based on DNA barcoding and chloroplast genome analysis.

Bupleuri Radix is the dry root of certain species of the genus Bupleurum and is commonly used in traditional Chinese medicine. The increasing global demand for Bupleuri Radix cannot be fulfilled with wild populations only. Therefore, cultivated Bupleurum is now the main commercial source of this medicinal product. Different species of Bupleurum show different medicinal properties and clinical effects, making reliable authentication and assignment of correct botanical origin for medicinal species critical. However, accurate identification of the cultivated Bupleurum species is difficult due to dramatic morphological variations resulting from cultivation. In this study, we sampled 56 cultivated Bupleurum populations of six different morphotypes (Types A-F) from the main production areas of China, and 10 wild populations of four species were used as reference materials. Conventional DNA barcoding was conducted to identify cultivated Bupleurum species. Additionally, verification based on complete chloroplast genomes was performed and new chloroplast markers were developed and evaluated. The combination of these methods resulted in the successful identification of all cultivated Bupleurum individuals. Three chloroplast regions are recommended as additional barcodes for the genus: ycf4_cemA, psaJ_rpl33, and ndhE_ndhG. This is a reliable and promising strategy that can be applied to the authentication of natural products and the identification of other medicinal plant species with similar taxonomic problems.

[Analysis and suggestions on harvest period standards of plant medicinal materials in 2020 edition of Chinese Pharmacopoeia].

The improvement of the harvest period standards is critical in the quality control of Chinese medicinal materials. The present study statistically analyzed the harvest period standards of plant medicinal materials in the 2020 edition of Chinese Pharmacopoeia(Vol.Ⅰ) and put forward the existing problems and suggestions based on herbal records and modern research to provide references for the improvement of the standards. According to the statistical analysis, in 499 types of plant medicinal materials, harvest period standards are recorded under 486 types, accounting for 97.4%, and are lacking in the remaining. Only one medicinal material(Stellariae Radix) is recorded with the standard of the harvest year. The standards of the harvest season and phenological period are recorded under 233 types, accounting for 46.7%. For 237 types, only harvest season is specified, accounting for 47.5%, and for 15 types, only harvest phenological period is specified, accounting for 3.0%. Among 222 types mainly derived from cultivation and 51 types from wild resources and cultivation, only 11 types are recorded with harvest period of cultivated products. Only Stellariae Radix is recorded with the harvest period standards for the wild and cultivated products separately. The harvest period standards of plant medicinal materials with different medicinal parts have certain rules to follow. The main problems about the harvest period standards are discovered. Specifically, no harvest period standards are recorded under 13 types of plant medicinal materials. Almost all perennial cultivated medicinal materials are not recorded with harvest year standard. No phenological period standard is found under 250 types of plant medicinal materials. There is no clear distinction between the harvest period standards of cultivated and wild products. The evidence for harvest period standards of 26 types of plant medicinal materials that can be harvested all year round is insufficient. As a result, it is proposed to strengthen basic research in response to the above-mentioned problems and improve the harvest period standards as soon as possible to ensure the quality of Chinese medicinal materials.

What's the choice for goji: Lycium barbarum L. or L. chinense Mill.?

ETHNOPHARMACOLOGICAL RELEVANCE: For over one millennium, goji berries have been used traditionally as food and medicine in eastern Asia. In recent decades, it has become increasingly popular globally. However, the biocultural development of goji is poorly known. The botanical origin of goji is controversial: in many but not all modern regional or international quality standards, L. barbarum is accepted exclusively as the botanical origin of goji. AIM OF THE STUDY: Focusing on historical, biogeographical, botanical, phytochemical and pharmacological data, the overarching aim is to understand the biological origin of goji's historical uses, as well as whether the two species can be used interchangeably. MATERIALS AND METHODS: The taxonomic literature on L. barbarum and L. chinense were analysed, followed by a study of botanical specimens and fieldwork. Historical herbals and gazetteers were employed to define the historical producing areas and medical properties of goji. An identification of the species used in history was carried out. In a final step the phytochemical and pharmacological literature on the species was compared. RESULTS AND DISCUSSION: Due to their morphological similarity and different accessibility, fruits of both L. barbarum and L. chinense have been used interchangeably as food and medicine at least since 682 CE. While the fruit of L. barbarum was recognized to be superior in quality, the fruit of L. chinense was commonly used as an equivalent because of its easier accessibility. Cultivation of L. barbarum in China since 1960s improved its availability, which likely lead to its exclusive use as source of goji in China. The long-term safe use with no reported major safety concerns supports that these two species both are useful sources for medicinal Lycium. CONCLUSIONS: Medicinal plants had been used traditionally long before they were named in scientific nomenclature system. Therefore, the understanding of traditional herbal knowledge and the adequate use of those traditional medicines require a reliable identification based on archival records. This study developed an approach for the identification of species used historically, with an integrated analysis of specimens, historical herbals, and national gazetteers. Additionally, their different chemical profiles and pharmacological activities indicate that they should not be used interchangeably. Further scientific evidence is required for their safe and effective use.

Comparison of anti-inflammatory effects of Lonicerae Japonicae Flos and Lonicerae Flos based on network pharmacology.

Objective: In Chinese herbal medicine (CHM) history, Lonicerae Japonicae Flos and Lonicerae Flos were used clinically as one drug, but now they are admitted as two herbal medicines in Chinese Pharmacopoeia (2010 edition). This study used network pharmacology to investigate whether the two can be used interchangeably for the treatment of inflammatory diseases in TCM clinical practice. Methods: Lonicerae Japonicae Flos and Lonicerae Flos were compared in the inflammation mechanism including core targets, Gene Ontology (GO), pathway and principle chemical components by the method of network pharmacology. Results: Lonicerae Japonicae Flos and Lonicerae Flos shared in six targets accounting for 66.7% of the entire core targets and more than half of the GO terms and pathways are similar. Organic acids are dominent compounds responsible for anti-inflammatory effects. Three of the compounds that bind to core targets including luteolin, quercetin and kaempferol, are shared in both herbs. Conclusion: Due to high similarity between Lonicerae Japonicae Flos and Lonicerae Flos, we believe that they can be used interchangeably for the inflammation in clinical treatment.

Tracing the Edible and Medicinal Plant Pueraria montana and Its Products in the Marketplace Yields Subspecies Level Distinction Using DNA Barcoding and DNA Metabarcoding.

Increased public awareness of nutritional and health issues has resulted in the increasing consumption of food and herbal products made from the root of Pueraria montana var. lobata (Willd.) Maesen & S. M. Almeida ex Sanjappa & Predeep (kudzu vine) and P. montana var. thomsonii (Benth.) M. R. Almeida. The famous herbal medicine Yufeng Ningxin, which is used to treat cardiovascular diseases, can be legally produced only using P. montana var. lobata. However, precise identification at the subspecies level is usually challenging when these products' ingredients lose their morphological characteristics after deep processing. Here, six herbarium specimens, 21 expert-identified original plant samples, 30 raw material samples, 10 food products and 12 herbal products were collected to test the subspecies-level authentication abilities of ITS2 sequences. The results showed that ITS2 sequences can distinguish P. montana var. lobata from P. montana var. thomsonii with stable single nucleotide polymorphism (SNP) sites. A total of 93.3% of raw material samples were consistent with the markings on their labels, but only 50% of Gegen Powder samples were made from P. montana var. lobata. High-quality ITS2 sequences were successfully obtained from nine of the 12 herbal products using Sanger sequencing. Substitution and fungal contamination were detected in 3 herbal products by further DNA metabarcoding, even though thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC) tests verified that the products met existing quality standards. This study demonstrated that DNA barcoding is a powerful tool for the identification of P. montana var. lobata and P. montana var. thomsonii at the subspecies level, and we conclude that DNA barcodes can be broadly applied to trace the raw materials of food and herbal products.

[Effects of different concentrations of nitrogen and phosphorus on growth and active components of Salvia miltiorrhiza].

With annual Salvia miltiorrhiza seedlings as experimental material, using "3414" optimal regression design recommended by the Ministry of Agriculture and regularly watered with nutrient solution, through the dynamic sampling of S. miltiorrhiza in different growing stages, and the growth index, dry weight of plant root and content of active components were measured. The potted experiments were applied to study the effects of different nitrogen and phosphorus ratios on the growth, dry matter accumulation and accumulation of active components of S. miltiorrhiza, in order to explore a compatible fertilization method of nitrogen and phosphorus ratio that are suitable for production and quality of S. miltiorrhiza. The results reported as follows：①High concentrations of nitrogen fertilizer was beneficial to dry matter accumulation of S. miltiorrhiza aerial parts, and low concentration of nitrogen fertilizer transferred the dry matter accumulation to underground, and N1P1 could make the transfer ahead of time;②Regression analysis showed that in the early growth stage (before early July), we could use the nitrogen and phosphorus as basic fertilizer at a concentration of 1.521,0.355 g•L⁻¹ respectively to promote the growth of S. miltiorrhiza and at a concentration of 2.281,0.710 g•L⁻¹ respectively to promote the dry matter accumulation of root (after mid-August);③Five kinds of active components of S. miltiorrhiza decreased with the increase of nitrogen concentration, and increased with the increase of the concentration of phosphate fertilizer. Nitrogenous fertilizer, phosphate fertilizer in N-P=2∶3 ratio was more suitable for the accumulation of salvianolic acids, in N-P=1∶2 ratio was more suitable for the accumulation of tanshinone.

[Sequence analysis and identification of a chloroplast matK gene in Rhei Rhizoma from different botanical origins].

Rhei Rhizoma is a Chinese medicine with multiple botanical origins. There is a problem to identify it with conventional methods. To compare the characteristics of chloroplast matK gene sequences of different Rheum species and authenticate inspected species, the matK gene sequences of different species from different origins were amplified, cloned, and sequenced. Genomic DNA of Rheum plants was extracted using modified DNA extracted Kit and matK gene sequences were analyzed by ContingExpress, DNAman and MEGA5.0. The length of matK gene sequences of Rheum palmatum, R. tanguticum and R. officinale were 1 518 bp containing 57 variable loci. According to the mutation sites, R. palmatum, R. tanguticum and R. officinale were divided into different genotypes separately. Based on the established method according to the loci 587, 707, 838, we successfully identified the genuine Rheum species from its adulterants.

[A new natural product from root of Angelica dahurica cv. Qibaizhi].

OBJECTIVE: To study the chemical constituents of the root of Angelica dahurica cv. Qibaizhi. METHOD: The compounds were isolated and purified by the methods of solvent extraction and chromatographic technique, and their structures were identified on the basis of the analyses of spectral data. RESULT: Twenty-seven compounds were obtained and identified as isoimperatorin (1), alloisoimperatorin (2), oxypeucedanin hydrate (3), byakangelicin (4), alloimperatorin (5), beta-sitosterol (6), dibutylphthalate (7), 6-(3,3-dimethylallyloxy) -7-hydroxycoumarin {7-hydroxy-6-[ (3-methyl-2-butenyl) oxy] -coumarin, 8}, xanthotoxol (9), 5-hydroxy-8-methoxypsoralen ( 5-hydroxyxanthotoxin, 10), smyrindiol (11), umbelliferone (12), bergapten (13), imperatorin (14), phellopterin (15), isoimpinellin (16), 6-acyl-7-methoxycoumarin (17), psoralen (18), isoscopletin (19), scoparone (20), xanthotoxin (21), angelol A (22), oxypeucedanin ethanolate (oxypeucedanin hydrate-3"-ethyl ether, 23), dehydrogeijerin (pablohopin, 24) , ulopterol (25), heraclenol (26), and pabulenol (27), respectively. CONCLUSION: Compound 8 was a new natural product and the above-mentioned all the compounds except for compounds 1, 3 and 14 were isolated from the root of A. dahurica cv. Qibaizhi for the first time.

[Study on effect of different processing methods on seven main chemical components of wild and cultivated Paeonia lactiflora].

OBJECTIVE: To study on the effect of different processing methods on the contents of seven major constituents in wild and cultivated Paeonia lactiflora, gallic acid, catechin, albiflorin, paeoniflorin, pentagalloylglucose, benzoic acid and paeonol, in order to provide reference basis for different efficacy and formation mechanism of Paeonia Radix Rubra and Paeonia Radix Alba. METHOD: Wild and cultivated P. lactiflora were dealt with by four processing methods, direct drying, drying after boiling, drying after decorticating and boiling, and drying after boiling and decorticating. HPLC was use to simultaneously determine the contents of seven chemical constituents. RESULT: Wild P. lactiflora showed notable higher content of paeoniflorin and catechin than cultivated P. lactiflora, whereas cultivated P. lactiflora showed higher content of albiflorin than wild P. lactiflora. Both of them were less affected by process methods in above three constituents. Drying after boiling, drying after decorticating and boiling, and drying after boiling and decorticating methods reduced the content of benzoic acid and paeonol to trace in both wild and cultivated P. lactiflora. Clustering analysis results showed that all processing methods assembled wild and cultivated P. lactiflora in 2 groups. CONCLUSION: The content differences of Paeonia Radix Rubra and Paeonia Radix Alba are mainly caused by their own differences and less affected by processing methods.

[Comparative study of plant morphological characteristics and photosynthetic physiological characteristics of wild and cultivated Paeonia lactiflora].

OBJECTIVE: The aim of this study was to compare the plant morphological characteristics and photosynthetic physiological characteristics of Paeonia Radix Rubra and Paeonia Radix Alba, and find evidence of whether there are intraspecific variations of Paeonia lactiflora. METHOD: With wild and cultivated P. lactiflora of the roots as asexual reproduction materials for cultivation experiment, morphological index, photosynthetic parameters and photosynthetic pigment content were measured. RESULT: In the 9 of morphological characteristics between wild and cultivated P. lactiflora, the number of leaf, blade length, the width of leaf and the length of top leaf showed a significant difference. The variances within wild P. lactiflora group were big than those within cultivated P. lactiflora in 8 of morphological characteristics. In photosynthetic physiological characteristics, wild P. lactiflora were significantly higher than cultivated P. lactiflora in Max. net photosynthetic rate (P(max)), light compensation point (LCP), light saturation point(LSP), CO2 saturation point (CSP) and CO2 saturated P(n) (P(m)). And significantly lower than cultivated P. lactiflora in maximum net photosynthetic rate (P(max)), light compensation point (LCP), light in apparent quantum yield (AQY), CO2 compensation point (CCP), carboxylation efficiency (CE) and photosynthetic pigment content. CONCLUSION: Wild and cultivated P. lactiflora have significant difference in morphological characteristics of leaf and photosynthetic physiological characteristics.

[Separation of 1-phenyl-3-methyl-5-pyrazolone derivatives of traditional Chinese medicine oligosaccharides by capillary zone electrophoresis].

A simple and practical capillary zone electrophoretic (CZE) method has been developed for the separation of traditional Chinese medicine oligosaccharides according to their relative molecular masses (M(r)). To optimize the conditions of the method, the concentration and pH value of the running buffer, and the applied voltage were evaluated. The optimized conditions were as follows: 1-phenyl-3-methyl-5-pyrazolone (PMP) derivatives of the oligosaccharides were separated with 50 mmol/L phosphate buffer (pH 2.5) as running buffer and the voltage was 15 kV. The detection was performed with an ultraviolet detector at 245 nm. An uncoated fused-silica capillary of 50 microm i. d. and 38/48 cm length (effective length/total length) was employed, and a hydrodynamic pressure injection (10 cm x 2 s) was applied. In order to examine the practicability of the method for the analysis of actual traditional Chinese medicine oligosaccharide samples, a complex sample consisting of some usual monosaccharides and oligosaccharides degraded from beta-cyclodextrin was separated under the electrophoretic conditions. And then, the method was applied to the analysis of the controlled degradation products of Indigowoad Root polysaccharide. The results indicated that the oligosaccharide sample could fully be separated from low to high M(r). This method is efficient and practical. In addition, the separation hypothesis of PMP derivatives of monosaccharides and oligosaccharides in pH 2.5 running buffer is also discussed, which would be helpful for us to understand the unusual migration of the PMP derivatives of rhamnose and mannose.

[Genetic variability and interrelationships of mainly quantitative traits in Glycyrrhiza uralensis cultivated population].

OBJECTIVE: The main aim of the research was to evaluate genetic variability and interrelationships of mainly quantitative traits in 2-year population, and provide a basis for high-yield breeding of Glycyrrhiza uralensis. METHOD: Four genotype G. uralensis population were transplanting in four different environment using complete randomized block design with three replication, and the 10 quantitative traits, including plant height (PH), stem diameter (SD), tiller number (TN), taproot length (TRL), root length (RL), root diameter (RD), diameter of 20 cm below the root head (D20), taperingness (TR), lateral root number (LRN) and root fresh weight (RFW) were measured in field. RESULT: The difference among population for all evaluated traits were significant (P<0.05) through Duncan's multiple range tests, and the coefficient of variation of RFW and LRN were above 25%. The analysis of variance was used to evaluate the traits of four populations across to four different environment Genotype, environment and their interaction effect were significant (P<0.05) or highly significant (P<0.01) for mainly evaluated traits. Simple correlation between traits showed that PH, SD, LRN, RL, RD and D20 had highly significant (P<0.01) and positive correlation with RFW. Results of the path coefficient analyses showed that D20 had the greatest positive direct effect on RFW, followed by the traits of PH and RL. CONCLUSION: Selection for increased D20, RL and PH would be the best indirect selection traits for increasing root yield. Meanwhile, ample genetic variability exists in the G. uralensis 2-year population, it could be used for breeding improvement of root yield.