Integrated transcriptomics and metabolomics analyses of the effects of bagging treatment on carotenoid biosynthesis and regulation of Areca catechu L.

Introduction: Fresh Aareca nut fruit for fresh fruit chewing commonly found in green or dark green hues. Despite its economic significance, there is currently insufficient research on the study of color and luster of areca. And the areca nut fruits after bagging showed obvious color change from green to tender yellow. In the study, we tried to explain this interesting variation in exocarp color. Methods: Fruits were bagged (with a double-layered black interior and yellow exterior) 45 days after pollination and subsequently harvested 120 days after pollination. In this study, we examined the the chlorophyll and carotenoid content of pericarp exocarp, integrated transcriptomics and metabolomics to study the effects of bagging on the carotenoid pathway at the molecular level. Results: It was found that the chlorophyll and carotenoid content of bagged areca nut (YP) exocarp was significantly reduced. A total of 21 differentially expressed metabolites (DEMs) and 1784 differentially expressed genes (DEGs) were screened by transcriptomics and metabolomics. Three key genes in the carotenoid biosynthesis pathway as candidate genes for qPCR validation by co-analysis, which suggested their role in the regulation of pathways related to crtB, crtZ and CYP707A. Discussion: We described that light intensity may appear as a main factor influencing the noted shift from green to yellow and the ensuing reduction in carotenoid content after bagging.

Molecular mechanisms of macrophage immunomodulation mediated by Areca inflorescence polysaccharides based on RNA-seq analysis.

The elucidation of the immunomodulatory molecular mechanisms of polysaccharides has contributed to their further development and application. In this study, the effect of Areca inflorescence polysaccharide (AFP2a) on macrophage activation was confirmed and the detailed mechanisms were investigated based on a comprehensive transcriptional study and specific inhibitors. The results showed that AFP2a induced macrophage activation (M1 polarization), promoting macrophage proliferation, reactive oxygen species production, nitric oxide and cytokine release, and costimulatory molecule expression. RNA-seq analysis identified 5919 differentially expressed genes (DEGs). For DEGs, GO, KEGG, and Reactome enrichment analyses and PPI networks were conducted, elucidating that AFP2a activated macrophages mainly by triggering the Toll-like receptor cascade and corresponding adapter proteins (TIRAP and TRIF), thereby resulting in downstream NF-κB, TNF, and JAK-STAT signaling pathway expression. The inhibition assay revealed that TLR4 and TLR2 were essential for the recognition of AFP2a. This work provides an in-depth understanding of the immunoregulatory mechanism of AFP2a while offering a molecular basis for AFP2a to serve as a potential natural immunomodulator.

Phytohormones and candidate genes synergistically regulate fruitlet abscission in Areca catechu L.

BACKGROUND: The fruit population of most plants is under the control of a process named "physiological drop" to selectively abort some developing fruitlets. However, frequent fruitlet abscission severely restricts the yield of Areca catechu. To reveal the physiological and molecular variations in this process, we detected the variation of phytohormone levels in abscised and non-abscised fruitlets in A. catechu. RESULTS: The levels of gibberellin acid, jasmonic acid, salicylic acid, abscisic acid and zeatin were elevated, while the indole-3-acetic acid and indole-3-carboxaldehyde levels were declined in the "about-to-abscise" part (AB) of abscission zone (AZ) compared to the "non-abscised" part (CK). Then the differentially expressed genes (DEGs) between AB and CK were screened based on transcriptome data. DEGs involved in phytohormone synthesis, response and transportation were identified as key genes. Genes related to cell wall biosynthesis, degradation, loosening and modification, and critical processes during fruit abscission were identified as role players. In addition, genes encoding transcription factors, such as NAC, ERF, WRKY, MADS and Zinc Finger proteins, showed differentially expressed patterns between AB and CK, were also identified as candidates. CONCLUSIONS: These results unraveled a phytohormone signaling cross talk and key genes involved in the fruitlet abscission process in A. catechu. This study not only provides a theoretical basis for fruitlet abscission in A. catechu, but also identified many candidate genes or potential molecular markers for further breeding of fruit trees.

Structural characterization of polysaccharide fractions in areca (Areca catechu L.) inflorescence and study of its immunological enhancement activity in vitro and in vivo.

To obtain the structure-function relationship of the polysaccharides derived from areca (Areca catechu L.) inflorescences in the aspect of its immunomodulatory ability, the plant-based polysaccharide was isolated and purified on column chromatography. The purity, primary structure and immune activity of four polysaccharide fractions (AFP, AFP1, AFP2 and AFP2a) were characterized comprehensively. The main chain of AFP2a was confirmed to be composed of â†’ 3,6)-ß-D-Galp-(1→, with branch chains linked to the O-3 position on the main chain. The immunomodulatory activity of the polysaccharides was evaluated using the RAW264.7 cells and immunosuppression mice model. It was observed that AFP2a enabled greater NO release (49.72 µmol/L) than other fractions, significantly promoted the phagocytic activity of macrophages, and improved splenocyte proliferation and T lymphocyte phenotype in mice. The present results may shine a light on a new research direction in immunoenhancers and provide a theoretical foundation for the development and application of areca inflorescence.

Areca catechu L. ameliorates chronic unpredictable mild stress-induced depression behavior in rats by the promotion of the BDNF signaling pathway.

OBJECTIVES: In this study, we have investigated the anti-depressant effects of the fruit Areca catechu L. (ACL) and elucidated its potential underlying mechanism using a rat model of chronic unpredictable mild stress (CUMS). METHODS: CUMS was induced in rats to establish a depression animal model for 28 days. According to the baseline sucrose preference, the male rats were divided into 6 different groups. They were treated with paroxetine hydrochloride, ACL, and water once a day until the behavioral tests were performed. The levels of corticosterone (CORT), malondialdehyde (MDA), catalase (CAT), and total superoxide dismutase (T-SOD) in serum were detected using a commercial kit, and the concentrations of 5-hydroxytryptamine (5-HT) and dopamine (DA) monoamine neurotransmitters in the brain tissues were detected by liquid chromatography-tandem mass spectrometry. doublecortin (DCX) expression in the hippocampal dentate gyrus (DG) was determined by immunofluorescence, and the relative abundance of brain-derived neurotrophic factor (BDNF), TrkB, PI3K, p-AKT/AKT, PSD-95, and p-GSK-3ß/GSK-3ß of brain tissues were assayed by western blot. RESULTS: ACL markedly increased sucrose preference, decreased the immobility time, and shortened the feeding latency of CUMS-induced rats. CUMS induction resulted in marked changes in the contents of the monoamine neurotransmitters (5-HT and DA) in the hippocampus and cortex of brain tissues and the levels of CORT, MDA, CAT, and T-SOD in serum, whereas ACL administration alleviated these considerable changes. ACL promoted DCX expression in DG and increased the protein levels of BDNF, TrkB, PI3K, p-AKT/AKT, PSD-95, and p-GSK-3ß/GSK-3ß in the brains of CUMS-induced rats. CONCLUSIONS: Our results indicated that ACL may improve depression-like behaviors in CUMS-induced rats by decreasing the hyperfunction and oxidative stress of the hypothalamic-pituitary-adrenal axis, stimulating hippocampal neurogenesis, and activating the BDNF signaling pathway.

Integrated Transcriptomic and Metabolomic Analyses Reveal the Molecular and Metabolic Basis of Flavonoids in Areca catechu L.

Areca catechu L., of the Arecaceae family, is widely distributed in tropical Asia. In A. catechu, the extracts and compounds, including flavonoids, have various pharmacological activities. Although there are many studies of flavonoids, the molecular mechanism of their biosynthesis and regulation remains unclear in A. catechu. In this study, 331 metabolites were identified from the root, stem, and leaf of A. catechu using untargeted metabolomics, including 107 flavonoids, 71 lipids, 44 amino acids and derivatives, and 33 alkaloids. The transcriptome analysis identified 6119 differentially expressed genes, and some were enriched in the flavonoid pathway. To analyze the biosynthetic mechanism of the metabolic differences in A. catechu tissues, 36 genes were identified through combined transcriptomic and metabolomic analysis, in which glycosyltransferase genes Acat_15g017010 and Acat_16g013670 were annotated as being involved in the glycosylation of kaempferol and chrysin by their expression and in vitro activities. Flavonoid biosynthesis could be regulated by the transcription factors, AcMYB5 and AcMYB194. This study laid a foundation for further research on the flavonoid biosynthetic pathway of A. catechu.

Genome-Wide Association Analysis of Fruit Shape-Related Traits in Areca catechu.

The areca palm (Areca catechu L.) is one of the most economically important palm trees in tropical areas. To inform areca breeding programs, it is critical to characterize the genetic bases of the mechanisms that regulate areca fruit shape and to identify candidate genes related to fruit-shape traits. However, few previous studies have mined candidate genes associated with areca fruit shape. Here, the fruits produced by 137 areca germplasms were divided into three categories (spherical, oval, and columnar) based on the fruit shape index. A total of 45,094 high-quality single-nucleotide polymorphisms (SNPs) were identified across the 137 areca cultivars. Phylogenetic analysis clustered the areca cultivars into four subgroups. A genome-wide association study that used a mixed linear model identified the 200 loci that were the most significantly associated with fruit-shape traits in the germplasms. In addition, 86 candidate genes associated with areca fruit-shape traits were further mined. Among the proteins encoded by these candidate genes were UDP-glucosyltransferase 85A2, the ABA-responsive element binding factor GBF4, E3 ubiquitin-protein ligase SIAH1, and LRR receptor-like serine/threonine-protein kinase ERECTA. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis showed that the gene that encoded UDP-glycosyltransferase, UGT85A2, was significantly upregulated in columnar fruits as compared to spherical and oval fruits. The identification of molecular markers that are closely related to fruit-shape traits not only provides genetic data for areca breeding, but it also provides new insights into the shape formation mechanisms of drupes.

Integrated metabolomic and transcriptomic analysis revealed the flavonoid biosynthesis and regulation in Areca catechu.

INTRODUCTION: Flavonoids are active substances in many herbal medicines, and Areca catechu fruit (AF), an important component in traditional Chinese medicine (TCM), is rich in flavonoids. Different parts of AF, Pericarpium Arecae (PA) and Semen Arecae (SA), have different medicinal effects in prescription of TCM. OBJECTIVE: To understand flavonoid biosynthesis and regulation in AF. METHODOLOGY: The metabolomic based on liquid chromatography-tandem mass spectrometry (LC-MS/MS) and the transcriptome based on high-throughput sequencing technology were combined to comprehensively analyse PA and SA. RESULTS: From the metabolite dataset, we found that 148 flavonoids showed significant differences between PA and SA. From the transcriptomic dataset, we identified 30 genes related to the flavonoid biosynthesis pathway which were differentially expressed genes in PA and SA. The genes encoding the key enzymes in the flavonoid biosynthesis pathway, chalcone synthase and chalcone isomerase (AcCHS4/6/7 and AcCHI1/2/3), were significantly higher expressed in SA than in PA, reflecting the high flavonoid concentration in SA. CONCLUSIONS: Taken together, our research acquired the key genes, including AcCHS4/6/7 and AcCHI1/2/3, which regulated the accumulation of flavonol in AF. This new evidence may reveal different medicinal effects of PA and SA. This study lays a foundation for investigating the biosynthesis and regulation of flavonoid biosynthesis in areca and provides the reference for the production and consumption of betel nut.

Areca catechu L.ameliorates chronic unpredictable mil stress-induced depression behavior in rats by the promotion of the BDNF signaling pathway / 中国药理学与毒理学杂志

OBJECTIVE To investigated the anti-de-pressant effects of the fruit Areca catechu L.(ACL)and elucidated its potential underlying mechanism using a rat model of chronic unpredictable mild stress(CUMS).METHODS CUMS was induced in rats to establish a depression animal model for 28 d.According to the baseline sucrose preference,the male rats were divided into six different groups.They were treated with parox-etine hydrochloride,ACL,and water once a day until the behavioral tests were performed.The levels of corticoste-rone(CORT),malondialdehyde(MDA),catalase(CAT),and total superoxide dismutase(T-SOD)in serum were de-tected using a commercial kit,and the concentrations of 5-hydroxytryptamine(5-HT)and dopamine(DA)mono-amine neurotransmitters in the brain tissues were detect-ed by liquid chromatography-tandem mass spectrometry.Doublecortin(DCX)expression in the hippocampal den-tate gyrus(DG)was determined by immunofluorescence,and the relative abundance of brain-derived neurotrophic factor(BDNF),TrkB,PI3K,p-Akt/Akt,PSD-95,and p-GSK-3β/GSK-3β of brain tissues were assayed by West-ern blotting.RESULTS ACL markedly increased sucrose preference,decreased the immobility time,and short-ened the feeding latency of CUMS-induced rats.CUMS induction resulted in marked changes in the contents of the monoamine neurotransmitters(5-HT and DA)in the hippocampus and cortex of brain tissues and the levels of CORT,MDA,CAT,and T-SOD in serum,whereas ACL administration alleviated these considerable changes.ACL promoted DCX expression in DG and increased the protein levels of BDNF,TrkB,PI3K,p-Akt/Akt,PSD-95,and p-GSK-3β/GSK-3β in the brains of CUMS-induced rats.CONCLUSION Our results indicated that ACL may improve depression-like behaviors in CUMS-induced rats by decreasing the hyperfunction and oxidative stress of the hypothalamic-pituitary-adrenal axis,stimulating hippo-campal neurogenesis,and activating the BDNF signaling pathway.

Advances in chemical constituents of Areca catechu L.and their effects on nervous system / 中国药理学与毒理学杂志

Areca nut is the dried and mature seed of Areca catechu L.in the palm family,which is a medicinal and food resource.It tastes bitter,spicy and warm.It has the functions of killing insects,eliminating accumulation,activating qi and nourishing water.It contains alkaloids,polyphenols,polysaccharides,fatty acids,amino acids,flavonoids,triterpenes and steroids and other chemical components.It has a certain protective effect on the ner-vous system,and mainly includes:① Arecoline con-tained in areca nut can stimulate the sympathetic nerve and simultaneously stimulate M receptor and N receptor,which can be used for the treatment of Alzheimer's dis-ease.② Antidepressant effect.Arecoline can easily pass through the blood-brain barrier and stimulate nerve cell receptors,thus promoting the body's excitement.Total phenols increase the content of monoamine transmitters in the brain by decreasing the content of monoamine oxi-dase,thus exerting therapeutic effect on depression.③ Antioxidant effect.Polyphenols and flavonoids can reduce oxidative damage of nervous system,and their antioxidant capacity is related to their content.④ Anti-inflammatory effect.Areca polysaccharides can exert anti-inflammatory activity by inhibiting the production of NO in macrophages induced by lipopolysaccharide.⑤Improved digestion.Areca nut water extract has a pro-moting effect on gastrointestinal motility,which is not only related to M choline receptor,but also probably related to α-adrenalin receptor.⑥Other effects.Such as anti-throm-bosis,anti-atherosclerosis,hypoglycemia,blood lipid reg-ulation,anti-tumor,anti-allergy,bacteriostasis,etc.This review summarized the chemical composition and phar-macological action of Areca catechu in recent years.It was found that the mechanism of action of Areca cate-chu L.is still unclear and further research is needed to provide basis for scientific utilization of Areca catechu.

Anticholinesterase activity of Areca Catechu: In Vitro and in silico green synthesis approach in search for therapeutic agents against Alzheimer's disease.

For many years, the primary focus has been on finding effective treatments for Alzheimer's disease (AD), which has led to the identification of promising therapeutic targets. The necessity for AD stage-dependent optimal settings necessitated a herbal therapy strategy. The plant species Areca Catechu L. (AC) was selected based on the traditional uses against CNS-related diseases. AC leaf extract were prepared using a Soxhlet extraction method and hydroxyapatite nanoparticles (HAp-NPs) were synthesized from the same (AC-HAp-NPs). Powder X-ray diffractometer (XRD), scanning electron microscopy (SEM), transmission electron microscopy (TEM), selected area electron diffraction (SAED) and fourier transform infrared spectroscopy (FTIR) were used to confirm the structure and morphology of the as-prepared AC-HAp-NPs. The crystalline character of the AC-HAp-NPs was visible in the XRD pattern. The synthesized material was found to be nanoflake, with an average diameter of 15-20 nm, according to SEM analysis. The TEM and SAED pictures also revealed the form and size of AC-HAp-NPs. In vitro anti-acetylcholinesterase and butyrylcholinesterase (AChE and BChE) activities of hydroxyapatite nanoparticles produced from an AC leaf extract was tested in this study. When compared to control, AC-HAp-NPs had higher anti-AChE and BChE activity. The anti-acetylcholinesterase action of phytoconstituents generated from AC leaf extract was mediated by 4AQD and 4EY7, according to a mechanistic study conducted utilizing in silico research. The global and local descriptors, which are the underpinnings of Conceptual Density Functional Theory (CDFT), have been predicted through the MN12SX/Def2TZVP/H2O model chemistry to help in the comprehension of the chemical reactivity properties of the five ligands considered in this study. The CDFT experiments are supplemented by the calculation of several useful calculated pharmacokinetics indices, their expected biological targets connected to the bioavailability of the five ligands in order to further the goal of studying their bioactivity.

Genome-Wide Identification of the DOF Gene Family Involved in Fruitlet Abscission in Areca catechu L.

Fruitlet abscission frequently occurs in Areca catechu L. and causes considerable production loss. However, the inducement mechanism of fruitlet abscission remains mysterious. In this study, we observed that the cell architecture in the abscission zone (AZ) was distinct with surrounding tissues, and varied obviously before and after abscission. Transcriptome analysis of the "about-to-abscise" and "non-abscised" AZs were performed in A. catechu, and the genes encoding the plant-specific DOF (DNA-binding with one finger) transcription factors showed a uniform up-regulation in AZ, suggesting a role of the DOF transcription in A. catechu fruitlet abscission. In total, 36 members of the DOF gene family distributed in 13 chromosomes were identified from the A. catechu genome. The 36 AcDOF genes were classified into nine subgroups based on phylogenic analysis. Six of them showed an AZ-specific expression pattern, and their expression levels varied according to the abscission process. In total, nine types of phytohormone response cis-elements and five types of abiotic stress related cis-elements were identified in the promoter regions of the AcDOF genes. In addition, histochemical staining showed that lignin accumulation of vascular bundles in AZ was significantly lower than that in pedicel and mesocarp, indicating the specific characteristics of the cell architecture in AZ. Our data suggests that the DOF transcription factors might play a role in fruitlet abscission regulation in A. catechu.

Genome-wide investigation of ZINC-IRON PERMEASE (ZIP) genes in Areca catechu and potential roles of ZIPs in Fe and Zn uptake and transport.

Iron (Fe) and Zinc (Zn) are essential nutrient elements for plant growth and development. Here, we observed the effects of Fe and Zn deficiency in seedlings of Areca catechu L. (areca palm), one of the most cultured palm trees in tropic regions. Results revealed that Fe deficiency causes strong chlorosis with the significantly decreased chlorophyll biosynthesis level and photosynthetic activities in the top third young leaf (L3) of seedlings. Zn deficiency caused light chlorosis in all three young leaves which slightly decreased chlorophyll biosynthesis and photosynthetic activities. Analysis of the Fe and Zn concentration in leaves and roots indicated that absorption and distribution of these two ions share cooperative pathways, since Zn deficiency caused Fe increasing, and vice versa. Therefore, we focused on the ZINC-IRON PERMEASE (ZIP) genes in areca trees. From the whole-genome data set we obtained, 6 ZIP genes were classified, and a phylogenetic tree was constructed with other 38 ZIP genes from model plants to find their potential functions. We also analyzed the expression pattern of AcZIP1-6 genes under Zn and Fe deficiency by transcriptomic approaches. With these results, we constructed an expression atlas of AcZIP1-6 genes in leaves and roots of areca seedlings with the dynamic expression levels under Fe and Zn deficient conditions. In conclusion, we provide evidence to understand the absorption and transport of nutrient elements, Fe and Zn, in the tropic agricultural plant A. catechu.

Iron Deficiency Leads to Chlorosis Through Impacting Chlorophyll Synthesis and Nitrogen Metabolism in Areca catechu L.

Deficiency of certain elements can cause leaf chlorosis in Areca catechu L. trees, which causes considerable production loss. The linkage between nutrient deficiency and chlorosis phenomenon and physiological defect in A. catechu remains unclear. Here, we found that low iron supply is a determinant for chlorosis of A. catechu seedling, and excessive iron supply resulted in dark green leaves. We also observed morphological characters of A. catechu seedlings under different iron levels and compared their fresh weight, chlorophyll contents, chloroplast structures and photosynthetic activities. Results showed that iron deficiency directly caused chloroplast degeneration and reduced chlorophyll synthesis in chlorosis leaves, while excessive iron treatment can increase chlorophyll contents, chloroplasts sizes, and inflated starch granules. However, both excessive and deficient of iron decreases fresh weight and photosynthetic rate in A. catechu seedlings. Therefore, we applied transcriptomic and metabolomic approaches to understand the effect of different iron supply to A. catechu seedlings. The genes involved in nitrogen assimilation pathway, such as NR (nitrate reductase) and GOGAT (glutamate synthase), were significantly down-regulated under both iron deficiency and excessive iron. Moreover, the accumulation of organic acids and flavonoids indicated a potential way for A. catechu to endure iron deficiency. On the other hand, the up-regulation of POD-related genes was assumed to be a defense strategy against the excessive iron toxicity. Our data demonstrated that A. catechu is an iron-sensitive species, therefore the precise control of iron level is believed to be the key point for A. catechu cultivation.

First Report of Grammothele lineata Causing Stem Rot of Areca catechu in Hainan Province, China.

Areca catechu L. (areca) belongs to the Arecaceae family, which is composed of 181 genera and 2,600 species (Christenhusz and Byng 2016), is cultivated extensively in Southern and Southeastern Asia (Peng et al. 2015). Areca has a long history for its important economic and medicinal benefits and is one of the most important commercial crops in Hainan province, China. In recent years, root rot and stem rot diseases have occurred, causing areca plants to wither and even die. The serious symptoms mainly appeared in the Hainan province (Li et al. 2006). In March 2018, the rotten tissues of the diseased plants were observed to become brittle, brown, and even black from the stem base to the root; the outer leaves turned yellow, dry, and dropping in areca plantations of Qionghai county. The disease can spread from individual plants to the whole plantation in one to two years, with the characteristics of large-scale occurrence and rapid transmission, causing huge economic losses. Diseased tissues (5 × 5 mm) were disinfected with 75% ethanol for 30 s, 1% HgCl2 for 1 min, washed in sterile water, placed on potato dextrose agar (PDA) medium and incubated at 28°C (Gao et al. 2019). Pure isolates were obtained by transferring the mycelium around the diseased tissues to PDA several times. The colonies were white and cottony after culturing for 7 days. The reverse side of the colony was yellowish white. Basidiospores were hyaline, thin-walled, smooth, 1.7-1.8 x 1.6-1.7 µm (n=30) in size and circular or ellipse in shape, in addition to a dimitic hyphal system (Das et al. 2017). For molecular identification, the genomic DNA of the isolate was extracted using the thermolysis method (Zhang et al. 2010). The ribosomal internal transcribed spacer (ITS) region was amplified using the primer pairs ITS1/ITS4 (White et al. 1990), and the amplified DNA fragments were sequenced. The obtained ITS sequence (GenBank accession No. MW534416) had 99.36% identity with the reference sequence (GenBank accession No. KX013157) of Grammothele lineata Berk. & M.A. Curtis. A phylogenetic tree was constructed with software MEGA7 using the neighbor-joining method, showing that the isolate was grouped in the same clade as G. lineata. To fulfil Koch's postulates, a pathogenicity test was performed using the stems of 6-month-old healthy areca seedlings. Stem surfaces were sterilized with 70% ethanol for 30 s, rinsed three times with sterile water, and gently stabbed with a sterile needle, and then inoculated with a 1-cm-diameter colonized PDA disk from a 7-day culture on wounds, moistened with wet cotton, and wrapped with a fresh plastic wrap. Plants inoculated with sterile PDA medium plugs were used as a control. The inoculation assay was carried out twice, with five plants in both control and treatment in each test. After 20 days, the stems of the plants inoculated with the pathogen exhibited rotten symptoms, and the leaves began to become yellow and shrunken, while the control plants had only the surface of the stems discolored slightly and the inner tissue was undamaged. The fungus was re-isolated from the infected stems. Based on the morphological observations and ITS sequence analysis, the isolate was identified as G. lineata. As far as we know, this is the first report of G. lineata causing the stem rot of areca in China.

Comparative anthelmintic efficacy of Arundo donax, Areca catechu, and Ferula assa-foetida against Haemonchus contortus / Eficácia anthelmíntica comparativa de Arundo donax, Areca catechu e Ferula assa-foetida contra Haemonchus contortus

Abstract In the present study, anthelmintic activities of Arundo (A.) donax L., Areca (Ar.) catechu L., and Ferula (F.) assa-foetida L. were determined. Leaves of A. donax L., latex of F. assa-foetida L. and seeds of Ar. catechu L. in different solvent fractions were subjected to in vitro (egg hatch assay; EHA, and adult motility assay; AMA) and in vivo (faecal egg count reduction test; FECRT) tests of anthelmintic activity using Haemonchus contortus model. In the AMA, crude aqueous methanol extracts (CAME) and ethyl acetate fractions of F. assa-foetida at 10 hr post-treatment showed maximum mortality of H. contortus at 12.5-50 mg mL-1. In the EHA, CAME of F. assa-foetida was identified as a potent ovicide based on its low LC50 (16.9 µg mL-1), followed in order by Ar. catechu and A. donax. Results from the FECRT also showed the extract of F. assa-foetida L. to be more effective than those of Ar. catechu L. and A. donax L., against the gastrointestinal parasitic nematodes. Chloroform and ethyl acetate fractions showed better anthelmintic activities against the adult worms in vitro, while CAME of these plants were better than their crude powders in vivo. It is recommended to document and investigate indigenous knowledge of possible medicinal plants to plan scientific trials that may justify their endorsement.

Resumo No presente estudo, as atividades anti-helmínticas de Arundo (A.) donax L., Areca (Ar.) Catechu L. e Ferula (F.) assa-foetida L. foram determinadas. Folhas de A. donax L., látex de F. assa-foetida L. e sementes de Ar. catechu L. em diferentes frações de solvente foram submetidos a testes in vitro (teste de eclosão de ovos, EHA e ensaio de motilidade em adultos, AMA); e in vivo (teste de redução da contagem de ovos fecais, FECRT) de atividade anti-helmíntica, usando-se Haemonchus contortus. Na AMA, extratos aquosos brutos de metanol (CAME) e frações de acetato de etila de F. assa-foetida. Dez horas pós-tratamento, apresentaram mortalidade máxima de H. contortus em 12,5-50 mg mL-1. No EHA, CAME de F. assa-foetida foi identificado como um ovicida potente baseado em seu baixo LC50 (16,9 µg mL-1), seguido em ordem por Ar. catechu e A. donax. Os resultados do FECRT também mostraram que o extrato de F. assa-foetida L. é mais eficaz do que o de Ar. catechu L. e A. donax L., contra nematoides parasitas gastrointestinais. As frações clorofórmio e acetato de etila mostraram melhores atividades anti-helmínticas contra vermes adultos in vitro, enquanto o CAME dessas plantas foi melhor do que o pó bruto in vivo. Recomenda-se documentar e investigar o conhecimento indígena de possíveis plantas medicinais para planejar ensaios científicos que possam justificar seu endosso.

Prevalence of Yellow Leaf Disease (YLD) and its Associated Areca Palm Velarivirus 1 (APV1) in Betel Palm (Areca catechu) Plantations in Hainan, China.

Yellow leaf disease (YLD) is an economically important disease affecting betel palm in several countries, the cause of which remains unclear despite associations with putative agents, including phytoplasmas. In this study, we screened the potential casual agents associated with YLD in Hainan, China using next-generation sequencing and revealed the association of areca palm velarivirus 1 (APV1) with the YLD-affected palm. The complete genome of the APV1-WNY isolate was determined to be 17,546 nucleotides in length, approximately 1.5 kb longer than the previously reported APV1_HN genome. Transmission electron microscopy showed that APV1 particles are flexuous and filamentous, a typical morphology of species in the Closteroviridae family. Comparison of symptomatic and symptomless tree populations showed a strong association between APV1 and YLD. APV1 was detected in Pseudococcus sp. mealybugs sampled from YLD-affected trees in many locations, suggesting that mealybugs are a potential transmission vector for APV1. Although further studies are needed to confirm a causal relationship, these results provide timely information for the prevention and management of YLD associated with APV1.

Discussion and consideration on pharmacology and toxicology of Areca catechu and its rational application / 中草药

Areca catechu ranks the first of China's precious "four southern medicines", with a history of more than 1 800 years of medicinal use and extensive clinical application. The pharmacological effects of A. catechu mainly focus on insect repellent, anti-tumor, anti-aging, anti-atherosclerosis and the effects on digestive system and nervous system. In recent years, safety problems such as oral cancer caused by consumption of A. catechu have been frequently reported, and the main chemical components of both medicinal and edible A. catechu are arecoline, so the clinical safety of medicinal A. catechu has been questioned. Based on the chemical composition, this paper analyzes the pharmacological and toxicological characteristics of A. catechu, discusses the points for attention in clinical application, in order to provide a reference basis for the clinical safe and rational use of A. catechu.

Research progress on chemical constituents and pharmacological activities of Areca catechu / 中草药

Areca catechu is a traditional Chinese medicine and an important medicine and food homologous variety, it’s widely distributed in tropical and subtropical provinces in southern China and other countries in South Asia and Southeast Asia. The fruitage of A. catechu is not only a kind of great fruit widely eaten to welcome guests, but also its peels and seeds can be used as medicines. Arecae Semen is one of the four precious “Southern Medicine” in China. Modern studies have shown that A. catechu contains alkaloids, tannins, flavonoids, terpenoids and other chemical constituents. It has multiple activities in promoting digestion, lowering blood pressure and anti-depressant, anti-oxidation, anti-inflammatory, anti-parasitic and bacteriostatic activities etc. In this paper, the research progress of chemical constituents and pharmacological activities of A. catechu in recent years was summarized, which provided research basis for the edible safety, research and development of traditional Areca products.

Screening and Identification of the Metabolites in Rat Plasma and Urine after Oral Administration of Areca catechu L. Nut Extract by Ultra-High-Pressure Liquid Chromatography Coupled with Linear Ion Trap-Orbitrap Tandem Mass Spectrometry.

Areca catechu L. nut, a well-known toxic traditional herbal medicine, has been widely used to treat various diseases in China and many other Asian countries for centuries. However, to date the in vivo absorption and metabolism of its multiple bioactive or toxic components still remain unclear. In this study, liquid chromatography coupled with tandem mass spectrometry was used to analyze the major components and their metabolites in rat plasma and urine after oral administration of Areca catechu L. nut extract (ACNE). A total of 12 compounds, including 6 alkaloids, 3 tannins and 3 amino acids, were confirmed or tentatively identified from ACNE. In vivo, 40 constituents, including 8 prototypes and 32 metabolites were identified in rat plasma and urine samples. In summary, this study showed an insight into the metabolism of ACNE in vivo, which may provide helpful chemical information for better understanding of the toxicological and pharmacological profiles of ACNE.