[Qualitative and quantitative study of flavonoids in Notoginseng Radix et Rhizoma based on UPLC-Q-TOF-MS and HPLC-DAD].

The flavonoids in Panax notoginseng were qualitatively analyzed by ultra-high performance liquid chromatography-quadrupole-time of flight mass spectrometry(UPLC-Q-TOF-MS), and the content of three main flavonoids in P. notoginseng of different specifications and grades collected from different habitats was determined by HPLC-DAD. Flavonoids and anthocyanins were analyzed by UPLC-Q-TOF-MS/MS in the positive and negative ion modes, respectively. Twelve flavonoid glycosides and one anthocyanin glycoside in P. notoginseng were identified, but no flavonoid aglycones were detected. Among them, 12 compounds were identified in the underground part of P. notoginseng for the first time and eight compounds were first reported in this plant. Moreover, six and four compounds were identified in the Panax genus and the Araliaceae family for the first time, respectively. A method for simultaneous determination of three flavonoids in P. notoginseng was established by HPLC-DAD. The content of flavonoids in 721 P. notoginseng samples of 124 specifications and grades collected from 20 different habitats was simultaneously determined. Among three flavonoids determined, the content of quercetin-3-O-(2″-ß-D-xylosyl)-ß-D-galactoside was the highest with the average content in the tested samples of 161.0 µg·g~(-1). The content of compounds quercetin-3-O-hexosyl-hexoside and kaempferol-3-O-pentosyl-hexoside was relatively low, with the average content of 18.5 µg·g~(-1)(calculated as quercetin-3-O-sophoroside) and 49.4 µg·g~(-1)(calculated as kaempferol-3-O-sangbu diglycoside). There were significant differences in flavonoids content of samples from different production area. The content of flavonoids in spring P. notoginseng was significantly lower than that in winter P. notoginseng when the other influencing factors such as production areas, germplasm resources, and cultivation conditions were fixed. As for P. notoginseng of different specifications, the flavonoid content in the part connecting the taproot and the aboveground stem was significantly higher than that in other parts. The results of large-scale data showed that the flavonoid content gradually increased with the increase in the number of heads. There were significant differences between the flavonoid content in most specifications and grades, especially the 20-head P. notoginseng and countless head P. notoginseng, whose content was significantly lower and significantly higher than that of other specifications and grades, respectively. This study provides a scientific basis for the study of the effective components and quality control of P. notoginseng from the perspective of flavonoids.

[Effects of natural organic soil amendments on quality of Panax notoginseng].

The effects of four natural organic soil amendments on the quality and pesticide residues of Panax notoginseng were investigated through field experiments and the suitable dosage ratio of each soil amendment was selected to provide a new idea for the pollution-free cultivation of P. notoginseng. The four natural organic soil amendments used in this study were Jishibao, Jihuo, Fudujing, and omnipotent nutrients, which were produced by mixed fermentation of aboveground parts of different plants, biological waste residue, and biochar. During the experiments, only four soil amendments were applied to P. notoginseng instead of any pesticides and fertilizers. The experiment was designed as four factors and three levels. There were three dosage gradients(low, medium, and high) for Jishibao(A), Jihuo(B), Fudujing(C), and omnipotent nutrients(D). When the dosage of one soil amendment changed, the do-sage of the other soil amendments remained medium. There were 10 groups in addition to the soil amendment-free group as control(CK). The results showed that the four soil amendments could significantly improve the growth environment of P. notoginseng and increase the seedling survival rate and saponin content of P. notoginseng. The seedling survival rates of the treatment groups increased by 8.24%-30.05% as compared with the control group. Furthermore, the content of pesticide residues in P. notoginseng was too low to be detected, and that of heavy metals in P. notoginseng was far lower than the specified content in the Chinese Pharmacopoeia(2020). The optimal effect was achieved at medium dosage for all the soil amendments with the highest content of saponins, high seedling survival rate, and significantly reduced heavy metals, such as lead, cadmium, arsenic, and mercury.

SMRT- and Illumina-based RNA-seq analyses unveil the ginsinoside biosynthesis and transcriptomic complexity in Panax notoginseng.

Panax notoginseng is one of the most widely used traditional Chinese herbs with particularly valued roots. Triterpenoid saponins are mainly specialized secondary metabolites, which medically act as bioactive components. Knowledge of the ginsenoside biosynthesis in P. notoginseng, which is of great importance in the industrial biosynthesis and genetic breeding program, remains largely undetermined. Here we combined single molecular real time (SMRT) and Second-Generation Sequencing (SGS) technologies to generate a widespread transcriptome atlas of P. notoginseng. We mapped 2,383 full-length non-chimeric (FLNC) reads to adjacently annotated genes, corrected 1,925 mis-annotated genes and merged into 927 new genes. We identified 8,111 novel transcript isoforms that have improved the annotation of the current genome assembly, of which we found 2,664 novel lncRNAs. We characterized more alternative splicing (AS) events from SMRT reads (20,015 AS in 6,324 genes) than Illumina reads (18,498 AS in 9,550 genes), which contained a number of AS events associated with the ginsenoside biosynthesis. The comprehensive transcriptome landscape reveals that the ginsenoside biosynthesis predominantly occurs in flowers compared to leaves and roots, substantiated by levels of gene expression, which is supported by tissue-specific abundance of isoforms in flowers compared to roots and rhizomes. Comparative metabolic analyses further show that a total of 17 characteristic ginsenosides increasingly accumulated, and roots contained the most ginsenosides with variable contents, which are extraordinarily abundant in roots of the three-year old plants. We observed that roots were rich in protopanaxatriol- and protopanaxadiol-type saponins, whereas protopanaxadiol-type saponins predominated in aerial parts (leaves, stems and flowers). The obtained results will greatly enhance our understanding about the ginsenoside biosynthetic machinery in the genus Panax.

[Effects of light intensity on photosynthetic capacity and light energy allocation in Panax notoginseng.] / å ‰å¼ºå¯¹ä¸‰ä¸ƒå ‰åˆèƒ½åŠ›åŠèƒ½é‡åˆ†é çš„å½±å“.

To explore the effects of light intensity on photosynthetic characteristics and light adaptation of the shade-demanding species Panax notoginseng, the responses of photosynthesis to photosynthetic photon flux density, CO2 and sunflecks in the two-year-old Panax notoginseng were investigated under different levels of light intensity (29.8%, 9.6%, 5.0%, 1.4% and 0.2% of full sunlight). Meanwhile, chlorophyll a fluorescence parameter and light energy partitioning were also recorded and calculated in the responsive process. P. notoginseng grown under 29.8% full sunlight (FL) had a lower apparent quantum yield (AQY), potential photochemical quantum yield (Fv/Fm) and potential photochemical activity (Fv/Fo), however, it had a higher maximum net photosynthetic rate (Pn max), maximum electron transport rate (Jmax), F/Fm', electron transport rate (ETR), photochemical quenching (qP) and the proportion of light energy allocated to photochemistry dissipation (ΦPS2), but the non-photochemical quenching (NPQ) was not the highest. There were no significant differences in Pn max, light compensation point (LCP), light saturation point (LSP), dark respiration rate (Rd) among 9.6%FL and 5.0% FL treatments, but these treatments had relatively higher values of NPQ, AQY, carboxylation efficiency (CE), maximum carboxylation rate (Vc max), Fv/Fm and Fv/Fo. In addition, the Pn max, CE, Vc max, Jmax, ETR,F/Fm', qP, NPQ and ΦPSII decreased with the decrease in light intensity from 5.0%FL to 0.2%FL, and the proportion of light energy allocated to fluorescent dissipation (Φf,d) were increased. Under 500 µmol·m-2·s-1 light-flecks inducting, the ΦPSⅡof P. notoginseng increased slowly with the extension of time except for the treatment of less than 5.0%FL, and under the circumstance of 1.4%FL and 0.2%FL, ΦPSⅡreached significantly a perfect result, moreover, Φf,dincreased rapidly. These results suggested that the enhancement in photosynthetic electron transport to use the light energy and the moderate photoinhibition of PS2 might avoid the irreversible oxidative damage of photosynthetic organization in P. notoginseng under high levels of light intensity. Moderate shading was beneficial to maintain its higher non-photochemical quenching ability. However, its photosynthetic capacity depressed and the proportion of light energy allocated to non-photochemical pathway increased obviously in excessive shading, and it easily caused a light oxidative damage.

[DNA marker-assisted selection of medicinal plants (â ) .Breeding research of disease-resistant cultivars of Panax notoginseng].

DNA marker-assisted selection of medicinal plants is based on the DNA polymorphism, selects the DNA sequences related to the phenotypes such as high yields, superior quality, stress-resistance and so on according to the technologies of molecular hybridization, polymerase chain reaction and high-throughput sequencing, and assists the breeding of new cultivars. This study bred the first disease-resistant cultivar of notoginseng "Miaoxiang Kangqi 1" using the technology of DNA marker-assisted selection of medicinal plants and systematic breeding. The disease-resistant cultivar of notoginseng contained 12 special SNPs based on the analysis of Restriction-site Associated DNA Sequencing (RAD-Seq). Among the SNP (record_519688) was related to the root rot-resistant characteristics, which indicated this SNP could serve as genetic markers of disease-resistant cultivars and assist the systematic breeding. Compared to the conventional cultivated cultivars, the incidence rate of root-rot and rust-rot in notoginseng seedlings decreased by 83.6% and 71.8%, respectively. The incidence rate of root-rot respectively declined by 43.6% and 62.9% in notoginseng cultivation for 2 and 3 years compared with those of the conventional cultivated cultivars. Additionally, the potential disease-resistant groups were screened based on the relative SNP, and this model enlarged the target groups and advanced the breeding efficiency. DNA marker-assisted selection of medicinal plants accelerated the breeding and promotion of new cultivars, and guaranteed the healthy development of Chinese medicinal materials industry.

[DNA marker-assisted selection of medicinal plants (â ¢)Evaluation of disease resistance of "Miaoxiang Kangqi 1" --a new cultivar of Panax notoginseng].

DNA marked-assisted selection of medicinal plants accelerated the breeding and promotion of new cultivars, and guaranteed the healthy development of Chinese medicinal materials industry. The first disease-resistant cultivar of notoginseng, namely "Miaoxiang Kangqi 1", served as the object of study. We evaluated the Kangqi's resistance of seeds, seedlings and root against the pathological bacteria (Fusarum oxysporum) of root rot. Compared to the traditional cultivars, the disease index of notoginseng seeds declined by 52.0% after inoculation for seven days; the death rate of seedlings and disease index of root respectively decreased by 72.1% and 62.4% after inoculation for 25 days. Additionally, the growth inhibition ratio of notoginseng seeds and seedlings declined after inoculation. The seeds, seedlings and roots of "Miaoxiang Kangqi 1" showed significantly resistant to root rot. The evaluation of disease-resistance of Kangqi provided the basis for the popularization of new cultivar and guaranteed the favoring conduct of notoginseng pollution-free cultivation.

Stability-increasing effects of anthocyanin glycosyl acylation.

This review comprehensively summarizes the existing knowledge regarding the chemical implications of anthocyanin glycosyl acylation, the effects of acylation on the stability of acylated anthocyanins and the corresponding mechanisms. Anthocyanin glycosyl acylation commonly refers to the phenomenon in which the hydroxyl groups of anthocyanin glycosyls are esterified by aliphatic or aromatic acids, which is synthetically represented by the acylation sites as well as the types and numbers of acyl groups. Generally, glycosyl acylation increases the in vitro and in vivo chemical stability of acylated anthocyanins, and the mechanisms primarily involve physicochemical, stereochemical, photochemical, biochemical or environmental aspects under specific conditions. Additionally, the acylation sites as well as the types and numbers of acyl groups influence the stability of acylated anthocyanins to different degrees. This review could provide insight into the optimization of the stability of anthocyanins as well as the application of suitable anthocyanins in food, pharmaceutical and cosmetic industries.

[Contents and the Correlations of the Total Anthocyanins and Saponins of the Vegetative Organs of Panax notoginseng with Purple and Green Aerial Stems at Different Growth Stages].

Objective: To study the accumulation characteristics of total anthocyanins and saponins in the vegetative organ of Panax notoginseng with purple and green aerial stems at different growth stages. Methods: The contents of the total anthocyanins and saponins of the leaves, aerial stems, rhizomes and the adventitious roots of one-, two- and three-year-old Panax notoginseng with purple and green aerial stems were determined by spectrophotometry. Results: From one-year-old to three-year-old Panax notoginseng, the total anthocyanin contents (TACs) of the leaves, aerial stems and all vegetative organs and the percentages of the aerial stems TACs to those of all vegetative organs of purple aerial stemmed Panax notoginseng plants, the percentages of the total saponin contents (TSCs) of the leaves and aerial stems to those of all vegetative organs of the purple and green aerial stemmed plants and the percentages of the leaves TACs to those of vegetative organs of the green aerial stemmed plants all decreased. The TACs of the leaves and all vegetative organs of the green aerial stemmed plants, the TACs of the rhizomes and their adventitious roots, the vegetative organ TSCs and the percentages of the TSCs of the rhizomes and their adventitious roots to those of all vegetative organs of the purple and green aerial stemmed plants, and the percentages of the TACs of the rhizomes and their adventitious roots to those of all vegetative organs of the purple aerial stemmed plants were increased. The increasing rates of the TACs of the aerial stems and all vegetative organs and the TSCs of the aerial stems, rhizomes and their adventitious roots and all vegetative organs of the purple aerial stemmed plants were always higher than those of the green aeria stemmed plants, but the increasing rates of the leaves TSCs were lower than those of the green aerial stemmed plants, and the content changes of the vegetative organ TACs and TSCs of the purple and green aerial stemmed plants exhibited different correlations. Additionally, the three-years-old plants, the TSCs of the vegetative organs of the purple aerial stemmed plants were all higher than those of the green aerial stemmed plants, but the TACs of the leaves and rhizomes and their adventitious roots were lower than those of the green aerial stemmed plants. Conclusion: The saponin-accumulating capacity of the vegetative organs of purple aerial stemmed Panax notoginseng plants is stronger than that of green aerial stemmed Panax notoginseng.

[Analysis of global ecology of Panax notoginseng in suitability and quality].

In this study, the Geographic Information System for Global Medicinal Plants(GMPGIS) was used to assess the global production and ecological adaptation of Panax notoginseng. Based on climate factors and soil types of P. notoginseng from 326 sampling sites, which cover both traditional and current major producing regions, as well as on the results of the ecological similarity computing analysis, we obtained the maximum ecological similarity areas for P. notoginseng worldwide. The results indicated that China was the most suitable ecological and cultivated area globally for P. notoginseng, accounting for more than 70% of the total cultivated area in the world. The United States, Brazil, Portugal, and other 22 countries also had a small amount of potential suitable producing area. China has eight potential suitable producing provinces, including Yunnan, Fujian, Guangxi, Guizhou, etc. The prediction is consistent with the new district of P. notoginseng reported in recent years, which verifies the accuracy of the prediction of GMPGIS. We conducted a literature analysis on resource regeneration and quality ecology on P. notoginseng, and summarized the cultivation, wild tending models, and effects of environmental factors on the quality of P. notoginseng. The results provide scientific basis for selection of P. notoginseng, as well as the introduction, cultivation, and production of P. notoginseng worldwide.

Bioinformatics analysis of the squalene synthase gene and the amino acid sequence in ginseng species.

The cDNA sequence, their structure, physical properties, signal peptide, hydrophobicity, hydrophilicity, subcellular localization domain of transmembrane domain and evolutionary relationship of encoded amino acid sequences were analyzed in squalene synthase of 9 species of ginseng plant using bioinformatics methods on GenBank. The results showed that the averaged similarity of squalene synthase cDNA sequence structure in Ginseng species was 96.245%, the similarity of the amino acid encoding sequence was 95.5%. The secondary structure prediction results showed that the amino acid sequence of 9 squalene synthase had α helix and random coil as the main components. After the phylogenetic analysis in 9 kinds of ginseng species, we found that they can be divided into two subfamilies. The analysis showed that plants, animals, yeasts belonged to different species, the homology was high within plant species and animal species. By analyzing the ginseng species squalene synthase and their encoding gene bioinformatics features, we can provide the theoretical reference for the squalene synthase gene cloning and the genetic manipulation.

Structure-activity relationships of anthocyanidin glycosylation.

This paper summarizes the main achievements about the structure-activity relationships of anthocyanidin glycosylation. Anthocyanidin glycosylation is the essential step of anthocyanin biosynthesis and also the prerequisite of the further modifications of anthocyanins, which is jointly characterized by the glycosylation site, the type and number of the glycosyl as well as the glycosidic bond type. It generally enhances the stability, results in the hypsochromic effect and blueing, decreases the bioavailability and anticancer activity, and decreases, increases, or does not change the antioxidant activity of the anthocyanidins or anthocyanins, which is synergetically determined by the glycosylation site and the type and number of the glycosyl. Thereinto, in nature, the blue hues caused by the glycosylation may also be reinforced by the formation of the anthocyanic vacuolar inclusions. This review could provide a reference for the research of the structure-optimizing and function-exploiting of anthocyanins.

[Contents of total anthocyanins and total saponins as well as composition of saponin monomers of Purple and Green Notoginseng Radix et Rhizoma].

OBJECTIVE: The contents of total anthocyanins and total saponins as well as the composition of saponin monomers of Purple and Green Notoginseng Radix et Rhizoma were studied to compare the medicinal quality and commercial values. METHODS: Three-year-old Notoginseng Radix et Rhizoma was selected as the research materials. The contents of total anthocyanins and total saponins were determined by spectrophotometry. The compositions of saponin monomers were monitored by HPLC. The significance of content differences was determined by variance analysis. RESULTS: The contents of total anthocyanins and total saponins of Purple Notoginseng Radix et Rhizomawere about 204.85% and 33.86% higher than those of Green Notoginseng Radix et Rhizoma respectively. The Purple and Green Notoginseng Radix et Rhizoma both contained five saponin monomers whose contents were as follows: ginsenoside Rg1 > ginsenoside Rb1 > notoginsenoside R1 > ginsenoside Rd > ginsenoside Re. The contents of notoginsenoside R1, ginsenoside Rd and ginsenoside Re of Purple Notoginseng Radix et Rhizoma were about 16.03%, 10.83% and 5.39% higher than those of Green Notoginseng Radix et Rhizoma respectively. However, the contents of ginsenoside Rg1 and ginsenoside Rb1 of Green Notoginseng Radix et Rhizoma were about 0.93% and 3.33% higher than those of Purple Notoginseng Radix et Rhizoma respectively. With respect to Green Notoginseng Radix et Rhizoma, the increase of the total anthocyanins in Purple Notoginseng Radix et Rhizoma reached a significant level, but the increases of total saponins, notoginsenoside R1, ginsenoside Rd and ginsenoside Re and the decreases of ginsenoside Rg1 and ginsenoside Rb1 did not. CONCLUSION: The total anthocyanins accumulation in Notoginseng Radix et Rhizoma implies the content increases of the total saponins, notoginsenoside R1, ginsenoside Rd and ginsenoside Re, and the slight decreases of ginsenoside Rg1 and ginsenoside Rb1 contents; but the type and relative contents of saponin monomers remain unchanged. The medicinal quality and commercial value of Purple Notoginseng Radix et Rhizoma are higher than those of Green Notoginseng Radix et Rhizoma.