Novel harmine derivatives as potent acetylcholinesterase and amyloid beta aggregation dual inhibitors for management of Alzheimer's disease.

In this study, a series of potential ligands for the treatment of AD were synthesised and characterised as novel harmine derivatives modified at position 9 with benzyl piperazinyl. In vitro studies revealed that the majority of the derivatives exhibited moderate to potent inhibition against hAChE and Aß1 - 42 aggregation. Notably, compounds 13 and 17d displayed potent drug - likeness and ADMET properties, demonstrating remarkable inhibitory activities towards AChE (IC50 = 58.76 nM and 89.38 nM, respectively) as well as Aß aggregation (IC50 = 9.31 µM and 13.82 µM, respectively). More importantly, compounds 13 and 17d showed exceptional neuroprotective effects against Aß1 - 42-induced SH - SY5Y damage, while maintaining low toxicity in SH - SY5Y cells. Further exploration of the mechanism through kinetic studies and molecular modelling confirmed that compound 13 could interact with both the CAS and the PAS of AChE. These findings suggested that harmine derivatives hold great potential as dual - targeted candidates for treating AD.

Modules in robust but low-efficiency phyllosphere fungal networks drive saponin accumulation in leaves of different Panax species.

BACKGROUND: The phyllosphere mycobiome plays a crucial role in plant fitness and ecosystem functions. The complex microbial ecological networks (MEN) formed by these fungi remain poorly understood, particularly with regard to their organization strategy and their contributions to plant secondary metabolites such as saponin. RESULTS: In this study, we constructed six MENs from leaf epiphytic and endophytic mycobiomes of three Panax species distributed in the northeast and southwest ends of mainland China. Hub nodes were absent in these MENs, which were significantly more complex, robust, and less efficient compared to random graphs (P < 0.05), indicating a hub-independent high-robustness strategy to maintain structural homeostasis. The important roles of specific MEN modules in shaping leaf saponin profiles of each Panax species were proved by multiple machine learning algorithms. Positive regulation modules (PRMs) of total saponin content were further identified, which exhibited more deterministic ecological assembly and comprised of highly connected nodes as well as higher proportion of plant-associated fungal guilds compared to other network members, indicating their tight links with host plant. The significant and direct effects (P < 0.05) of PRMs on total saponin accumulation were validated by well-fitted structural equation models (χ2 < 0.3, P > 0.5). Taxonomic analysis revealed that Pleosporales and Chaetothyriales were significantly overrepresented by positive regulation taxa (PRT) of total saponin content (FDR < 0.05). Across PRT identified in three Panax species, Epicoccum and Coniothyrium were conservatively present, while species-specific taxa such as Agaricales were also found, indicating the conservatism and specificity of plant-fungi interactions associated with leaf saponin accumulation in Panax genus. CONCLUSIONS: These findings provide a foundation for understanding mechanisms maintaining the steady state of phyllosphere mycobiome in healthy plant, and offer clues for engineering phyllosphere mycobiome to improve the accumulation of bioactive secondary metabolites on the basis of network modules.

Comprehensive analysis of NAC transcription factors in Scutellaria baicalensis and their response to exogenous ABA and GA3.

The NAC is a plant-specific family of transcription factor that plays important roles in various biological processes. Scutellaria baicalensis Georgi, belongs to the Lamiaceae family and has been widely used as a traditional herb with a wide range of pharmacological activities, including antitumor, heat-clearing, and detoxifying functions. However, no study on the NAC family in S. baicalensis has been conducted to date. In the present study, we identified 56 SbNAC genes using genomic and transcriptome analyses. These 56 SbNACs were unevenly distributed across nine chromosomes and were phylogenetically divided into six clusters. Cis-element analysis identified plant growth and development-, phytohormone-, light-, and stress-responsive elements were present in SbNAC genes promoter regions. Protein-protein interaction analysis was performed using Arabidopsis homologous proteins. Potential transcription factors, including bHLH, ERF, MYB, WRKY, and bZIP, were identified and constructed a regulatory network with SbNAC genes. The expression of 12 flavonoid biosynthetic genes was significantly upregulated with abscisic acid (ABA) and gibberellin (GA3) treatments. Eight SbNAC genes (SbNAC9/32/33/40/42/43/48/50) also exhibited notable variation with two phytohormone treatments, among which SbNAC9 and SbNAC43 showed the most significant variation and deserved further study. Additionally, SbNAC44 displayed a positive correlation with C4H3, PAL5, OMT3, and OMT6, while SbNAC25 had negatively correlated with OMT2, CHI, F6H2, and FNSII-2. This study constitutes the first analysis of SbNAC genes and lays the basis foundation for further functional studies of SbNAC genes family members, while it may also facilitate the genetic improvement of plants and breeding of elite S. baicalensis varieties.

Dynamic analysis of physiological indices and transcriptome profiling revealing the mechanisms of the allelopathic effects of phenolic acids on Pinellia ternata.

Pinellia ternata (Thunb.) is a famous traditional Chinese medicine with high medicinal value, but its culture is strongly hindered by the continuous cropping obstacles (CCO) which are tightly associated with allelopathic effects. Deciphering the response mechanisms of P. ternata to allelochemicals is critical for overcoming the CCO. Here, we elucidate the response of P. ternata to phenolic acids treatment via physiological indices, cellular approaches, and transcriptome analysis. Phenolic acids showed a significant effect on the growth of P. ternata seedlings, similar to the phenotype of continuous cropping. Cellular analysis demonstrated that phenolic acids remarkably induced root cell death. Physiological analysis revealed that phenolic acids induced the overaccumulated of H2O2 and O 2 - in root cells. However, two exogenous antioxidants (L-ascorbic acid and ß-gentiobiose) aid in the scavenging of over-accumulated H2O2 and O 2 - by promoting the antioxidant enzyme activity such as superoxide dismutase (SOD), ascorbate peroxidase (APX), and catalase (CAT). Transcriptome analysis demonstrated that differentially expressed genes (DEGs) related to the cell wall degeneration and reactive oxygen species (ROS) metabolism were upregulated by phenolic acid treatment. In addition, downregulated DEGs involved in sucrose and starch metabolism and phenylpropanoid biosynthesis pathways decreased the key metabolites contents. Taken together, phenolic acids caused root cell death by inducing the overaccumulation of H2O2 and O 2 - , and L-ascorbic acid and ß-gentiobiose effectively alleviated ROS stress. The present study elucidates the underlying mechanism of the allelopathic effect of phenolic acids, offers valuable information for further understanding the mechanism of CCO, and could contribute to improving guidance for further P. ternata production.

Transcriptome and HPLC Analysis Reveal the Regulatory Mechanisms of Aurantio-Obtusin in Space Environment-Induced Senna obtusifolia Lines.

Senna obtusifolia is a famous medicinal plant that is widely used in Asian countries. Its seed plays an important role in the treatment of many diseases because it contains various anthraquinones and flavonoids. Our previous studies have indicated that three space environment-induced S. obtusifolia lines (SP-lines) i.e., QC10, QC29, and QC46, have higher seed yield and aurantio-obtusin (AO) content. However, the underlying mechanism of higher AO content in SP-lines is still unknown. Herein, transcriptome sequencing and HPLC were employed to analyze the differences between SP-lines and ground control (GC3) and elucidate the regulatory mechanisms of AO accumulation in SP-lines. The results show that 4002 differentially expressed genes (DEGs) were identified in SP-lines versus (vs.) GC3. DEGs in the QC10 vs. GC3, QC29 vs. GC3, and QC46 vs. GC3 comparisons were classified into 28, 36, and 81 GO terms and involved in 63, 74, and 107 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. KEGG pathway and gene expression analysis revealed that DEGs involved in anthraquinone pathways were significantly elevated in QC10 and QC46. Integrating the results of GO annotation, KEGG enrichment, and gene expression analysis, we propose that the elevated genes such as DAHPS, DHQS, and MenB enhance the metabolic flux in the anthraquinone pathway and promote AO content in QC10 and QC46. Taken together, this study elucidated the mechanism of AO content in SP-lines and provides valuable genetic information for S. obtusifolia. In addition, to the best of our knowledge, this study presents the first transcriptome analysis of environment-induced medicinal plants and paves the way to select elite S. obtusifolia varieties in the future.

Nitrate Increases Cadmium Accumulation in Sweet Sorghum for Improving Phytoextraction Efficiency Rather Than Ammonium.

Sweet sorghum has potential for phytoextraction of cadmium (Cd) owning to its large biomass and relatively high Cd tolerance. Nitrogen affects both growth and Cd concentrations in plants. However, different forms of nitrogen effects on Cd accumulation in sweet sorghum to improve efficiency of Cd phytoremediation is still elusive. In this study, nitrate substantially promoted both dry weight and Cd concentrations in leaves, stems + sheaths and roots of sweet sorghum when compared with ammonium. As a result, Cd accumulation in nitrate-supplied sweet sorghum was around 3.7-fold of that in ammonium-supplied plants under unbuffered pH condition, while the fold was about 2.2 under buffered pH condition. We speculated pH values and Cd species in the growth medium to some extent contributed to increased Cd accumulation as affected by nitrate. Net photosynthesis rate and Fv/Fm of nitrate-treated plants under Cd stress were higher than that of ammonium-treated plants when the pH was unbuffered. Responses of antioxidant capacity in roots to Cd stress with nitrate application were stronger than that with ammonium supplementation. Taken together, nitrate is more suitable than ammonium for Cd phytoextraction by using sweet sorghum, which is able to enhance at least double efficiency of phytoextraction.

Pinellia ternata (Thunb.) Breit: A review of its germplasm resources, genetic diversity and active components.

ETHNOPHARMACOLOGICAL RELEVANCE: The medicinal plant Pinellia ternata has been widely used in China, Korea, and Japan and has been demonstrated to be highly effective for treating cough, vomiting, infection, and inflammatory diseases. Modern pharmacological investigations have demonstrated its multiple activities, such as antitussive, expectorant, antiemetic, antitumor, antibacterial, and sedative-hypnotic activities. AIM OF THE REVIEW: This review aims to summarize the information about the biological traits, genetic diversity, active components, and continuous cropping obstacle of P. ternata in order to improve its use. MATERIALS AND METHODS: In this review, the relevant literature was gathered by using Pinellia ternata, genetic diversity, active components, and continuous cropping obstacle as the keywords from Google Scholar, PubMed, Springer Link, the Wiley online library, SciFinder, SCOPUS, Baidu Scholar, China national knowledge infrastructure (CNKI), and WANFANF DATA (up to April 2020). RESULTS: P. ternata is the most widely used herb in the Pinellia genus to treat several diseases. The genetic diversity of P. ternata has been extensively studied, and its high genetic diversity level in China has been demonstrated. Modern pharmacological research has indicated that amino acids, alkaloids, and polysaccharides are the main active components supporting P. ternata's medicinal effects. However, an efficient method for determining its active components is still unavailable. The method used to evaluate Pinelliae Rhizoma (PR) quality standards should be further optimized. The continuous cropping obstacle has a significant effect on the quantity and quality of P. ternata. The underlying mechanism of the continuous cropping obstacle needs to be further explored. CONCLUSIONS: P. ternata has emerged as a valuable source of traditional medicine. Some uses of P. ternata in medicine have been validated by pharmacological investigations. However, a more efficient analytical method should be established to evaluate the quality of PR based on multiple quality markers. Furthermore, high-performance liquid chromatography (HPLC) and DNA barcoding should be introduced to identify the authenticity of PR. In addition, the genes involved in the metabolic synthesis pathways of the main active components, population genetic relationships, the quality control of processed PR, and the continuous cropping obstacle need to be further elucidated. We hope this review will allow for better utilization of this valuable herb.

SmGRAS1 and SmGRAS2 Regulate the Biosynthesis of Tanshinones and Phenolic Acids in Salvia miltiorrhiza.

Salvia miltiorrhiza is one of the most widely used traditional Chinese medicinal plants because of its excellent performance in treating heart diseases. Tanshinones and phenolic acids are two important classes of effective metabolites, and their biosynthesis has attracted widespread interest. Here, we functionally characterized SmGRAS1 and SmGRAS2, two GRAS family transcription factors from S. miltiorrhiza. SmGRAS1/2 were highly expressed in the root periderm, where tanshinones mainly accumulated in S. miltiorrhiza. Overexpression of SmGRAS1/2 upregulated tanshinones accumulation and downregulated GA, phenolic acids contents, and root biomass. However, antisense expression of SmGRAS1/2 reduced the tanshinones accumulation and increased the GA, phenolic acids contents, and root biomass. The expression patterns of biosynthesis genes were consistent with the changes in compounds accumulation. GA treatment increased tanshinones, phenolic acids, and GA contents in the overexpression lines, and restored the root growth inhibited by overexpressing SmGRAS1/2. Subsequently, yeast one-hybrid, dual-luciferase, and electrophoretic mobility shift assays (EMSA) showed SmGRAS1 promoted tanshinones biosynthesis by directly binding to the GARE motif in the SmKSL1 promoter and activating its expression. Yeast two-hybrid assays showed SmGRAS1 interacted physically with SmGRAS2. Taken together, the results revealed that SmGRAS1/2 acted as repressors in root growth and phenolic acids biosynthesis but as positive regulators in tanshinones biosynthesis. Overall, our findings revealed the potential value of SmGRAS1/2 in genetically engineering changes in secondary metabolism.

Physiological, transcriptional, and metabolic alterations in spaceflight-subjected Senna obtusifolia.

Senna obtusifolia is a widely used medicinal herb in Asian countries. To select elite cultivars, S. obtusifolia seeds were carried by "ShenZhou Ⅷ" recoverable satellite to space. Three spaceflight-subjected lines (SP-lines), namely QC10, QC29, QC46, and their ground control line (GC-line) were cultivated on the ground. Previous studies demonstrated that biological traits and secondary metabolites are different between SP-lines and GC-line. Here, we combined physiological, transcriptional, and metabolic studies to compare the differences between SP-lines and GC-line. The results showed that activities of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), ascorbate peroxidase (APX), and monodehydroascorbate reductase (MDHAR) were dramatically increased in SP-lines as compared to that of GC-line. Transcript levels of SOD, POD, CAT, APX, and MDHAR were significantly up-regulated in SP-lines. Malondialdehyde (MDA) and hydrogen peroxide (H2O2) contents decreased in SP-lines. Seed yields of QC29 and QC46 were considerably higher than that of GC-line. Besides, QC29 had significantly higher aurantio-obtusin content. Pearson correlation coefficient analysis revealed positive relationships between POD and aurantio-obtusin, as well as APX and aurantio-obtusin. In conclusion, SP-lines have higher antioxidant gene expression level and antioxidant enzyme activity as compared to that of GC-line. With higher seed yield and aurantio-obtusin content, QC29 can be used to breed elite S. obtusifolia cultivars. This study provides a new insight in SP-lines and paves the way to breed elite S. obtusifolia cultivars in the future.

Remediation of deterioration in microbial structure in continuous Pinellia ternata cropping soil by crop rotation.

Pinellia ternata is a traditional Chinese herb that suffers from continuous cropping (CC), which significantly decreases both yield and quality. The influence of CC on the microbiome in P. ternata rhizosphere and the effects of remediation on microbiota by rotational cropping (CR) were assessed by Illumina high-throughput sequencing technology. CC tends to decrease the α-diversities as a function of cultivation time, whereas CR tends to increase them. Differentially abundant analysis showed that microbial structure was important in maintaining the health status of P. ternata rhizosphere. Results suggested that CC soils were mainly enriched for Pseudomonas, Rhizobium, and Streptophyta operational taxonomic units (OTUs), while the CR soils were mainly enriched for Rhizobium, Pseudomonas, Flavobacterium, Sphingomonas, Rhizobacter, and Arthrobacter OTUs. On the basis of the community dissimilarities, we grouped all sample replicates into three post hoc clusters in which soils were defined as healthy, health-suppressed, and health-depressed soils. The three soil types represented different soil physicochemical properties. The activities of the microbiome features, including ammonia oxidizer, sulfate reducer, nitrite reducer, dehalogenation, xylan degrader, sulfide oxidizer, nitrogen fixation, atrazine metabolism, chitin degradation, degraded aromatic hydrocarbons, and chlorophenol degradation, were also considerably different among the three soils.

Identification of seeds based on molecular markers and secondary metabolites in Senna obtusifolia and Senna occidentalis.

BACKGROUND: Senna obtusifolia and Senna occidentalis (Leguminosae), whose seeds have similar appearance and chemical constituents, are easily confused in using their seeds. To elucidate the similarities and differences between S. obtusifolia seeds and S. occidentalis seeds, three molecular markers and high performance liquid chromatography (HPLC) were employed to evaluate the seeds characteristics of these two medicinal herbs. RESULTS: The results showed that selected 3 ISSR and 7 SCoT primers could distinguish S. obtusifolia seeds from S. occidentalis seeds based on the specific band and UPGMA dendrogram. ITS2 sequence indicated that the intra-specific similarity of 20 S. obtusifolia and 16 S. occidentalis was 99.79 and 100.0%, respectively, while the inter-specific similarity between S . obtusifolia and S. occidentalis was 89.58%. Although phylogenetic analysis revealed that these two species had a close relationship, they were assigned to different branches. HPLC fingerprint results showed that seeds of S. obtusifolia and S. occidentalis shared some secondary metabolites, but aurantio-obtusin was not detected in S. occidentalis seeds which could differentiate S. obtusifolia seeds from S. occidentalis seeds. CONCLUSIONS: The present study not only compared the seeds characters of S. obtusifolia and S. occidentalis from molecular and secondary metabolites levels, but also provided a convenient method to identify S. obtusifolia seeds and S. occidentalis seeds effectively.

Spaceflight-induced variation on biological traits and effective components of Cassia obtusifolia.

The dry seeds of Cassia obtusifolia were carried by the "ShenZhou 8" satellite and sowed after landing. Based on our pri- or study on SP1, the characteristics of plants growth, physiological index and content of effective components were examined. The results showed that the QC10, QC29 strains matured 5 d earlier compared with control. The plant height, across diameter and ground diameter of QC10, QC29, QC46 strains was superior to the control at whole growth period. The branch number increased ranging from 4 to 11 and the number of pods reached 321, 313,281, respectively, which was dramatically higher than the control (246). The yield of QC10, QC29, QC46 strains increased noticeably from 31.4 to 63.2 g. The 1000-seed-weight of QC10, QC29, QC46 strains was 25.86, 25.88, 24.06 g, while the control was 23.69 g. Compared to the control, the mass fraction of chlorophyll was enhanced 1.098, 1.016, 0.297 mg. There was no significant difference in aurantio-obtusin and chrysophanol content of seeds. Through two years research, three high-yield mutant strains were obtained. This study indicates that spaceflight-induced mutants could provide new germplasm for C. obtusifolia breeding and offers the theoretical basis for further utilization of spaceflight-induced mutation to breed high-quality C. obtusifolia strains.