The bioactivity-guided isolation of potentially active natural products has been widely utilized in pharmaceutical discovery. In this study, by screening fungal extracts against coxsackievirus B3 (CVB3), three new aspochalasins, templichalasins A‒C (1‒3), along with six known aspochalasins (4‒9) were isolated from an active extract derived from the endophytic fungus Aspergillus templicola LHWf045. Compound 1 features a unique 5/6/5/7/5 pentacyclic ring system, while compounds 2 and 3 possess unusual 5/6/6/7 tetracyclic skeletons. Their structures were characterized through extensive spectroscopic analyses, electronic circular dichroism (ECD) calculations, and single-crystal X-ray diffraction analysis. Additionally, we demonstrated that compound 4 can be readily converted into compounds 1‒3 under mild acidic conditions and proposed a plausible mechanism for this conversion. Bioactivity evaluation of compounds 1‒9 against CVB3 revealed the inhibitory effects of all compounds against the virus. Notably, compound 9 exhibited superior antiviral activity, surpassing the commercial drug ribavirin in selectivity index (SI) value.
Antiviral Agents/isolation & purification* ; Aspergillus/chemistry* ; Molecular Structure ; Enterovirus B, Human/drug effects* ; Endophytes/chemistry* ; Cytochalasins/isolation & purification* ; Drug Discovery ; Humans

This study explored the growth-promoting effect and mechanism of the endophytic bacterium Kocuria rosea on Rehmannia glutinosa, aiming to provide a scientific basis for the development of green bacterial fertilizer. R. glutinosa 'Jinjiu' was treated with K. rosea, and the shoot parameters including leaf length, leaf width, plant width, and stem diameter were measured every 15 days. After 120 days, the shoots and roots were harvested. The root indicators(root number, root length, root diameter, root fresh weight, root dry weight, root volume, and root vitality) and secondary metabolites(catalpol, rehmannioside A, rehmannioside D, verbascoside, and leonuride) were determined. The R. glutinosa growth-promoting mechanism of K. rosea was discussed from the effect of K. rosea on the nutrient element content in R. glutinosa and rhizosphere soil and the genome information of this plant. After application of K. rosea, the maximum increases in leaf length, leaf width, plant width, and stem diameter were 35.67%(60 d), 25.39%(45 d), 40.17%(60 d), and 113.85%(45 d), respectively. The root number, root length, root diameter, root volume, root fresh weight, root dry weight, and root viability increased by 41.71%, 45.10%, 48.61%, 94.34%, 101.55%, 147.61%, and 42.08%, respectively. In addition, the content of rehmannioside A and verbascoside in the root of R. glutinosa increased by 76.67% and 69.54%, respectively. K. rosea promoted the transformation of nitrogen(N), phosphorus(P), and potassium(K) in the rhizosphere soil into the available state. Compared with that in the control, the content of available N(54.60 mg·kg~(-1)), available P(1.83 μmol·g~(-1)), and available K(83.75 mg·kg~(-1)) in the treatment with K. rosea increased by 138.78%, 44.89%, and 14.34%, respectively. The content of N, P, and K in the treatment group increased by 293.22%, 202.63%, and 23.80% in the roots and by 23.60%, 107.23%, and 134.53% in the leaves of R. glutinosa, respectively. K. rosea carried the genes related to colonization(rbsB, efp, bcsA, and gmhC), N, P, and K metabolism(narG, narH, narI, nasA, nasB, GDH2, pyk, aceB, ackA, CS, ppa, ppk, ppk2, pstS, pstA, pstB, and pstC), and indole-3-acetic acid and zeatin synthesis(iaaH and miaA). Further studies showed that K. rosea could colonize the roots of R. glutinosa and secrete indole-3-acetic acid(3.85 μg·mL~(-1)) and zeatin(0.10 μg·mL~(-1)). In summary, K. rosea promotes the growth of R.ehmannia glutinosa by enhancing the nutrient uptake, which provides a theoretical basis for the development of plant growth-promoting microbial products.
Rehmannia/metabolism* ; Endophytes/metabolism* ; Plant Roots/growth & development* ; Micrococcaceae/genetics* ; Data Mining ; Plant Leaves/metabolism* ; Genomics ; Rhizosphere

Aims: Endophytic fungi are the remarkable category of host-associated fungal community that invades the intercellular regions of host tissues, benefiting their host while obtaining an advantage. Fungal endophytes have lately attracted prominence as a source of active secondary metabolites. This investigation aimed to identify fungal endophytes that reside inside the leaves and stems of Aquilaria malaccensis. Methodology and results: Healthy A. malaccensis stems and leaves samples were collected. Clean leaves and stems were cut to a size of 1 cm, followed by sterilization using 75% ethanol for 1 min, 3% sodium hypochlorite solution for 1 min, and finally, rinsing with sterile water 3 times for 1 min and drying with sterile paper. The sterile samples were put onto Potato Dextrose Agar (PDA) media containing chloramphenicol for 7-14 days until the mycelium grew for morphological identification under a light microscope. Five endophytic fungi were recovered from leaves, while nine endophytic fungi were obtained from stems. Using morphological approaches, nine of the endophytes had observed to produce conidia fungi, whereas the others did not. Neopestalotiopsis sp., Aspergillus sp., Arthrinium sp., Curvularia sp., Podospora sp., Mucor sp. and Verticillium sp. were identified as nine of the fourteen endophytes. Conclusion, significance and impact of study The number of endophytic fungi discovered in different organs varies. Not all endophytic fungi that grow can create sexual phages. Six genera of endophytic fungi were identified.
Endophytes ; Thymelaeaceae

Two new polyketides, lasobutone A(1) and lasobutone B(2), along with three known compounds, guignardianone C(3), guignardic acid(4), and 4-hydroxy-17R-methylincisterol(5), were isolated from the endophytic fungi Xylaria sp. by silica gel, MCI, and preparative HPLC, which was separated from the Chinese medicinal material Coptis chinensis and cultivated through solid fermentation with rice. Their structures were elucidated on the basis of spectroscopic methods, such as MS, NMR, IR, UV, and ECD. Compounds 2 and 4 showed inhibitory activities against the nitric oxide(NO) production in the LPS-induced macrophage RAW264.7 with IC_(50) values of 58.7 and 42.5 μmol·L~(-1) respectively, while compound 5 exhibited cytotoxic activities against HT-29 with IC_(50) value of 14.3 μmol·L~(-1).
Antineoplastic Agents ; Coptis chinensis ; Endophytes/chemistry* ; Fungi ; Polyketides/chemistry*

Aims: This study was aimed to screen and isolate soil and endophytic fungi with the ability to biosynthesize stable silver nanoparticles (SNPs) with antimicrobial and antiproliferative activities. Methodology and results: A total of 60 fungal isolates isolated from soil and plant samples were screened for their ability to biosynthesize SNPs. Among which, 21 isolates have supported the biosynthesis of SNPs. Furthermore, the endophytic isolate PRR2.1 synthesized highly thermostable SNPs with long shelf life. The PRR2.1 isolate was identified as Albifimbria verrucaria by morphological and molecular means. The synthesis of SNPs was initially monitored by UV-Vis spectroscopy. Further characterization by transmission electron microscopy, X-ray diffraction and dynamic light scattering revealed well-dispersed spherical crystalline in nature SNPs with a mean size of 14 nm and zeta potential of –24.47 mV. Fourier transform infrared spectroscopy showed the presence of biomolecules adsorbed on the surface of biosynthesized SNPs responsible for their synthesis and stability. The mycosynthesized SNPs exhibited stronger antifungal activity against pathogenic strains of Aspergillus niger, A. flavus, A. fumigatus and Candida albicans with respect to its antibacterial activity against Escherichia coli, Staphylococcus aureus, Bacillus cereus and Klebsiella pneumoniae compared to standard antifungal itraconazole and antibiotic cefadroxil with mostly consistent minimum inhibitory concentration of 5.31 μg/mL. The biosynthesized SNPs demonstrated dose-dependent in vitro antiproliferative activity against cancerous HeLa cell line with IC50 value of 2.52 μg/mL and less cytotoxic activity against WI-38 (normal human lung fibroblasts) cell line with CC50 value of 10.2 μg/mL. Conclusion, significance and impact of study These results show the potential of endophytic fungi biosynthesized SNPs as possible biofriendly, safe and efficient antimicrobial agents with promising antiproliferative activity and low cytotoxicity, which can be furtherly implemented in various biomedical and biotechnological applications.
Silver ; Nanoparticles ; Soil Microbiology ; Endophytes ; Anti-Infective Agents ;

Aims: The objective of this study was to analyze the genome of endophytic actinomycete associated with orchids and evaluate its plant hormone activities, including phytohormone, siderophore, ammonia production, zinc and phosphate solubilization. Methodology and results: Strain DR1-2 isolated from the roots of the Thai orchid, Dendrobium christyanum Rchb.f., was closely related to Pseudonocardia alni DSM 44104T, P. antarctica DSM 44749T and P. carboxydivorans Y8T (99.93-100% similarity) based 16S rRNA gene sequence. This strain exhibited IAA production (294.10 ± 12.17 μg/mL), phosphate solubilization (2.20 ± 0.08 solubilization Index, SI), positive for siderophore production and ammonia production (36.99 ± 2.24 μg/mL). It showed a maximum IAA of 489.73 ± 8.90 μg/mL, when optimized using 0.5% Ltryptophan, pH 6 and incubated at 30 °C for 7 days. The IAA of strain enhanced the root length, shoot length, number of roots and fresh weight of rice seedlings (Oryza sativa L. cv. RD49). The draft genome of strain DR1-2 was 6,077,423 bp in 23 contigs with G+C content of 74.6%. The average nucleotide identity-Blast (ANIb) and average nucleotide identity-MUMmer (ANIm) values of strain DR1-2 and related type strains were 95.81 to 97.25% and the digital DNA-DNA hybridization (dDDH) values were 72.60 to 74.00%, respectively. Genomic analysis of strain DR1-2 revealed that the gene encodes the enzyme involved in the phytohormones biosynthesis and gene clusters involved in the biosynthesis of bioactive metabolites. Conclusion, significance and impact of study Endophytic actinomycete, Pseudonocardia strain DR1-2 from Thai orchid, D. christyanum Rchb.f., exhibited significant IAA production and affected the growth of the plant, which was the potential source of plant hormones for agricultural applications.
Endophytes ; Actinobacteria ; Pseudonocardia

Aims: Endophytic fungi are a diverse group of microorganisms that stay asymptomatically in the healthy tissues of the host. Many fungal endophytes are associated with the tea plant (Camellia sinensis) and the pathogens of the tea plant have the potential to grow as endophytes or act as latent pathogens during the initial growth of the plant. The present study aimed at screening tea endophytic fungi with the potential for control of a few critical phytopathogens, Fusarium sp., Lasiodiplodia theobromae, Pestalotiopsis sp. and Sclerotinia sclerotiorum while evaluating the efficiency of growth inhibition of these phytopathogens by the endophytic fungal isolates using in vitro assays. Methodology and results: Five endophytic fungal strains; Colletotrichum gloeosporioides, C. siamense, Daldinia eschscholtzii, Pseudopestalotiopsis chinensis and Phyllosticta capitalensis isolated from leaves of C. sinensis in Sri Lanka were evaluated for growth inhibition against plant pathogens; Fusarium sp., L. theobromae, Pestalotiopsis sp. and S. sclerotiorum using the dual culture assay and volatile compound-mediated inhibition assay. All the fungal endophytes used in this study exhibited antifungal activity against Fusarium sp., Pestalotiopsis sp. and S. sclerotiorum in the dual culture assay on PDA. Daldinia eschscholtzii (67.30%) and C. gloeosporioides (61.54%) showed strong antagonistic activity against S. sclerotiorum, while P. capitalensis (42.30%) demonstrated moderate activity. All the endophytic fungal strains showed moderate antifungal activities against Fusarium sp. The percentage growth inhibitions of Pestalotiopsis sp. by all the endophytic isolates tested were below 25.00%. In the volatile compound-mediated inhibition assay, none of the endophytic isolates showed visible inhibition against the phytopathogens used. Conclusion, significance and impact of study In this study, the fungal endophytes which showed potential antagonistic activity against the tested phytopathogens can be used to develop commercialized products of fungal biocontrol agents (BCAs) for controlling plant fungal diseases.
Endophytes ; Tea

In order to reveal the distribution and population characteristics of endophytic fungi from Zanthoxylum nitidum and the antibacterial potential,this study performed molecular identification and analyzed the genetic diversity and antibacterial activity of endophytic fungi from Z. nitidum in Guangxi. Through culture and molecular identification,35 strains,belonging to 15 genera,12 families,10 orders,4 classes,and 2 phyla,were isolated from various tissues of Z. nitidum,of which Colletotrichum and Fusarium were the dominant genera,respectively accounting for 20% of total strains. The diversity of endophytic fungi was significantly different among roots,stems,and leaves,as manifested by the significantly higher Shannon index( H') in stems( 1. 678) than in roots( 0. 882 1) and leaves( 0. 515 4). The antimicrobial activity analysis showed that 14. 28% of endophytic fungi inhibited at least one indicator pathogen. Among them,Fusarium sp. ZN-34 and Fusarium sp. ZN-26 separately demonstrated the strongest inhibitory effect on Escherichia coli and Staphylococcus aureus. In general,Fusarium sp. ZN-26 and Phialemoniopsis plurioloculosa ZN-35 were advantageous in suppressing the two bacteria owing to the broad spectrum and strong efficacy. In summary,Z. nitidum in Guangxi boasts rich endophytic fungi with the majority showing strong antibacterial activity,which can be used as candidates for the extraction and separation of basic antibacterial substances and the development of natural antibacterial agents.
Anti-Bacterial Agents/pharmacology* ; Anti-Infective Agents ; China ; Colletotrichum ; Endophytes/genetics* ; Fungi/genetics* ; Genetic Variation ; Humans ; Microbial Sensitivity Tests ; Zanthoxylum

The research on endophytes of medicinal plants mainly relies on the traditional culture and isolation methods. Because of their functions such as promoting host growth, improving stress resistance, promoting the accumulation of medicinal active ingredients or directly producing medicinal active ingredients, the endophytes of medicinal plants have gradually attracted wide attention. However, it was found that the strains isolated by traditional methods were not the true dominant endophytes of medicinal plants by comparing the results of traditional culture isolation with high-throughput sequencing. The blind and random nature of traditional methods leads to the lack of standards in terms of medium selection, culture time and interaction between species. On the contrary, high-throughput sequencing technology is an emerging molecular biology technology developed in recent decades. Due to its high resolution level and indepen-dent culture, it can be used for thorough analysis of the community structure and diversity of environmental microorganisms. Therefore, we proposed the strategy of using high-throughput sequencing technology to guide the traditional culture and isolation of endophytes from medicinal plants. Firstly, the endophytic structure and diversity of medicinal plants were completely clear by high-throughput sequencing technology, and the dominant endophytes of the host were unequivocal. Then according to the characteristics of each dominant endophytes design or query suitable medium for its growth to culture and isolation. Finally, the function of the isolates was studied. This method can prevent researchers from missing out on the important functional strains of the host, expand the research scope of endophytes of medicinal plants, and facilitate the in-depth excavation and utilization of endophytes of medicinal plants.
Endophytes/genetics* ; High-Throughput Nucleotide Sequencing ; Plants, Medicinal ; Research Design

The endophytes of medicinal plants play an important role in promoting the quality formation of the host. Therefore, this paper made a review of endophytes of medicinal plant Atractylodes lancea. According to previous studies, A. lancea boasts endophytes, such as fungi, bacteria, and actinomycetes, among which the beneficial microorganisms help the growth and development of A. lancea. There is a close interaction between the volatile oil of A. lancea and endophytes. Different endophytes vary in regulating the composition and content of the volatile oil of A. lancea, which might contribute to the quality formation of A. lancea. However, the information of the endophytic flora of A. lancea obtained by traditional culture and isolation is not enough to reflect the real situation of the endophytes of A. lancea. Little is known about the endophytes of A. lancea from different chemical types and different habitats, which is not conducive to the study of the ecological relationship between A. lancea and endophytes and limits the development and utilization of the endophytes. Therefore, at the end of this paper, the authors put forward suggestions for future research on endophytes in A. lancea, including:(1)mining the core endophyte resources of A. lancea by combining high-throughput sequencing with traditional culture and isolation;(2)exploring the relationship between the diversity of endophytes and chemical types of A. lancea;(3)strengthening the application of endophytes in A. lancea cultivation, in order to facilitate the cultivation efficiency and quality of A. lancea.
Atractylodes ; Endophytes ; Fungi ; Oils, Volatile ; Plants, Medicinal