Isolation and screening of fungi for enhanced agarwood formation in Aquilaria sinensis trees.

Agarwood is a resinous heartwood of Aquilaria sinensis that is formed in response to mechanical wounding. In the present study pre-treatment of Aquilaria sinensis was carried out, and then the dominant fungi were isolated and purified from the surface and electroshock holes of trees. The isolated Trichoderma sp. and Neurospora sp. were then screened for resistance against benzyl acetone and then inoculated into healthy Aquilaria sinensis trees. After six months, the agarwood was collected for analysis. The chemical composition of incense was analyzed using gas chromatography-mass spectroscopy, and 82 chemical constituents were identified. Agarwood products formed by using Trichoderma sp. and Neurospora sp. consisted of 50.22% and 48.71% ether extracts, respectively, which surpassed the 10% threshold specified by the Chinese Pharmacopoeia. Similarly, relative aromatic contents in the two agarwood products were 30.1% and 32.86%, while proportions of sesquiterpene constituents were 10.21% and 11.19%, respectively. These two agarwood-specific chemical constituents accounted for a large proportion of the total chemical composition, which showed that the generated agarwood was of good quality. The results of the study revealed that both Trichoderma sp. and Neurospora sp. were able to effectively induce agarwood production in Aquilaria sinensis trees in 6 months. This study expands the library of fungi that promote the production of agarwood from Aquilaria sinensis trees.

Diaporchalasins A-E, New Cytochalasins from the Endophytic Fungus Diaporthe sp. BMX12 Isolated from Aquilaria sinensis.

Five new cytochalasins, diaporchalasins A-E (1-5), together with 14 known congeners (6-19) were isolated from the endophytic fungus Diaporthe sp. BMX12, which was isolated from the branches of Aquilaria sinensis. The structures of the new compounds were elucidated by extensive spectroscopic analyses including high-resolution electron spray ionization mass spectrometry (HR-ESI-MS) and nuclear magnetic resonance (NMR). Their absolute configurations were assigned by theoretical electronic circular dichroism (ECD) calculations. Compounds 11 and 12 featuring a keto carbonyl at C-21 displayed cytotoxicity toward K562, BEL-7402, SGC-7901, A549, and HeLa cell lines with IC50 values ranging from 4.4 to 47.4 µM.

Fusarium oxysporum infection-induced formation of agarwood (FOIFA): A rapid and efficient method for inducing the production of high quality agarwood.

Agarwood, a non-wood product from the endangered Aquilaria and Gyrinops tress, is highly prized for its use in fragrances and medicines. The special formation process of agarwood is closely related to external injury and fungal infection. In this study, we demonstrate that infection of Aquilaria sinensis by Fusarium oxysporum, a soilborne fungus that causes vascular wilt diseases in diverse plants, induces agarwood formation. Based on these findings, an efficient method, termed F. oxysporum infection-induced formation of agarwood (FOIFA), was developed for the rapid production of quality agarwood. The agarwood formed in response to F. oxysporum infection was similar in structure and chemical composition to wild agarwood according to TLC (Thin-layer chromatography), HPLC (high performance liquid chromatography), and GC-MS (gas chromatography-mass spectrometry) analyses, except that the contents of alcohol-soluble extract, chromones, and essential oils (mainly sesquiterpenes) were higher in the formed agarwood.

Isolation and Characterization of Bacteria and Fungi Associated with Agarwood Fermentation.

Agarwood oil, often called attar, is a valuable perfume retainer derived from Aquilaria malaccensis. It is obtained through a process of fermentation followed by distillation. The microbes involved in fermentation during agarwood processing have neither been documented nor their role in the process deciphered yet. The present study characterizes the fungi and bacteria engaged in the agarwood fermentation and assesses their colony forming units (CFU) in three categories of agarwood fermentation broth (AFB) collected from three small-scale industries of Hojai, Assam at varying time points. It was interesting to note that grade-A AFB contained the highest CFU count for both bacteria and fungi; the metabolite profile also revealed that the oil obtained from grade-A agarwood comprised of the highest number of compounds. The CFU count increased from 0 to 21 days during fermentation. A total of 105 bacteria and 9 fungi were isolated from 3 different grades of AFB. Shannon index (H' = 0.81) was observed maximum in grade-C AFB and fisher index (α = 2.6) observed maximum for grade-A AFB. The genus Bacillus with a Pi value of 0.61 exhibited dominance among isolated bacteria, while the genus Galactomyces was dominant among fungi with a Pi value of 0.43. The metabolite profiles of three grades of oil obtained after fermentation and one solvent extracted (S.E.) grade agarwood oil analyzed using GC-MS, which showed distinct differences among the oil. The outcomes of this study are expected to create new opportunities for improving oil production methods by modulating biochemical processes involved in fermentation.

Integrating Multiple Omics Identifies Phaeoacremonium rubrigenum Acting as Aquilaria sinensis Marker Fungus to Promote Agarwood Sesquiterpene Accumulation by Inducing Plant Host Phosphorylation.

The present study aimed to explore the factors that promote persistent agarwood accumulation. To this end, we first investigated the morphological changes and volatile compound distribution in five layers of "Guan Xiang" agarwood. The agarwood-normal transition layer (TL), an essential layer of persistent agarwood accumulation, showed clear metabolic differences by microscopy and GC-MS analysis. Microbiome analysis revealed that Phaeocremonium rubrigenum was the predominant biomarker fungus in the TL of "Guan Xiang" agarwood samples. Among the seven isolated fungi, P. rubrigenum exhibited a significantly heightened ability to induce the production in Aquilaria sinensis seedlings, especially for sesquiterpene. Tracing the proteome profile changes in P. rubrigenum-induced A. sinensis calli for 18 ds showed that the fungus-induced sesquiterpene biosynthesis increased mainly through the mevalonate (MVA) pathway. Specifically, the phosphorylation modification level, instead of the protein abundance of transcription factors (TFs), showed corresponding changes during sesquiterpene biosynthesis, thus indicating that induced phosphorylation is the key reason for enhanced sesquiterpene production. IMPORTANCE Agarwood is an expensive resinous portion derived from Aquilaria plants and has been widely used as medicine, incense, and perfume. The factors involved in steady agarwood accumulation remain elusive. Our current study suggests that as a TL marker fungus, P. rubrigenum could persistently promote agarwood sesquiterpene accumulation by inducing phosphorylation of the TFs-MVA network in A. sinensis. Moreover, our work provides strategies to improve agarwood industry management and sheds light on the potential molecular mechanisms of plant adaptation to native microbial conditions.

Production of Volatile Compounds by a Variety of Fungi in Artificially Inoculated and Naturally Infected Aquilaria malaccensis.

Aquilaria malaccensis, the resinous agarwood, is highly valued in the perfumery and medicinal industry. The formation of fragrant agarwood resin inconsistently by various fungi is still not clearly understood. The current study investigated the agarwood quality and fungal diversity in artificially inoculated and naturally infected A. malaccensis. The chemical analysis of volatile compounds of agarwood was performed using the Solid Phase Micro Extraction (SPME) method, and the identification of fungi was made through a morphological observation using a light microscope. Gas chromatography analysis revealed the presence of essential compounds related to high-quality agarwood, such as 4-phenyl-2-butanone, ß-selinene, α-bulnesene, and agarospirol in both artificially inoculated and naturally infected agarwood but with some differences in the abundance. Further studies on the fungi associated with agarwood volatile compounds formation showed a total of ten fungal group isolates, which were identified based on morphological and molecular studies. The study revealed that agarwood from both artificial and natural sources were naturally infected with Fusarium, Botryosphaeria, Aspergillus, Schizophyllum, Phanerochaete, Lasiodiplodia, Polyporales, and Ceriporia species. This study has offered a potential opportunity to research further the promising development of fungal strains for artificial inducement of high-quality agarwood formation from A. malaccensis trees.

Saprophytic Bacillus Accelerates the Release of Effective Components in Agarwood by Degrading Cellulose.

The value of Agarwood increases with time due to the gradual release of its major components, but the mechanism behind this remains unclear. Herein we reveal that the potential driving force of this process is the degradation of cellulose in Agarwood by its saprophytic Bacillus subtilis. We selected 10-year-old Agarwood from different places and then isolated the saprophytic bacteria. We confirmed these bacteria from different sources are all Bacillus and confirmed they can degrade cellulose, and the highest cellulase activity reached 0.22 U/mL. By co-cultivation of the bacterium and Agarwood powder, we found that three of the strains could release the effective components of Agarwood, while they had little effect in increasing the same components in living Aquilaria sinensis. Finally, we demonstrated that these saprophytic Bacillus subtilis have similar effects on Zanthoxylum bungeanum Maxim and Dalbergiaod orifera T. Chen, but not on Illicium verum Hook. f, Cinnamomum cassia Presl and Phellodendron chinense Schneid. In conclusion, our experiment revealed that the saprophytic Bacillus release the effective components of Agarwood by degrading cellulose, and we provide a promising way to accelerate this process by using this bacterial agent.

Endophytic bacterial community of Stellera chamaejasme L. and its role in improving host plants' competitiveness in grasslands.

Stellera chamaejasme has become a problematic weed in northern and south-western grasslands of China. To evaluate a possible role of endophytes in its strong competitive capacity, the endophytic bacterial community of S. chamaejasme was investigated by culture-dependent and independent methods, and the growth-promoting traits of some culturable isolates as well as the benefit of endophyte ST3CS3 (Brevundimonas sp.) on host plants growth were studied. The results showed that 823 OTUs were generated with a 97% similarity level in the culture-independent study. They were classified into 29 phyla, 61 classes, 147 orders, 237 families and 440 genera. Among them, Pseudomonas and Ralstonia were the most dominant genera in belowground parts (G) (64.25%) and aboveground parts (S) (26.54%) respectively. The diversity and species richness of endophytes in S were significantly higher than that of G (P < 0.001, t-test). Contrary to this, the number of culturable bacteria in S was a little lower than that of G (P > 0.05, t-test). Totally, 176 isolates belonging to 30 morphotypes were obtained in the culture-dependent study. Among them, Acinetobacter was the most dominant genus in G (51.30%), then followed by Pseudomonas (6.09%) and Brevundimonas (6.09%), while Lysinibacillus (21.31%) was the most dominant genus in S, followed by Pseudomonas (11.48%). Growth-promoting trait tests indicated that 93.65% of the tested isolates (63) exhibited nitrogen-fixing, IAA-synthesizing, phosphorus or potassium solubilizing capacity, in which 77.97% belonged to Proteobacteria, a phylum found to contain more active isolates. Pot experiments demonstrated that endophyte ST3CS3 can significantly improve host plants growth and increase its nitrogen and chlorophyll content (P < 0.01, t-test). Therefore, we suggest that strong competitiveness of S. chamaejasme may in part be due to possession of high ratios of plant growth-promoting proteobacterial endophytes such as Pseudomonas, Acinetobacter and Brevundimonas.

Aromatic Polyketides from a Symbiotic Strain Aspergillus fumigatus D and Characterization of Their Biosynthetic Gene D8.t287.

The chemical investigation of one symbiotic strain, Aspergillus fumigatus D, from the coastal plant Edgeworthia chrysantha Lindl led to the isolation of eight compounds (1-8), which were respectively identified as rubrofusarin B (1), alternariol 9-O-methyl ether (2), fonsecinone D (3), asperpyrone A (4), asperpyrone D (5), fonsecinone B (6), fonsecinone A (7), and aurasperone A (8) by a combination of spectroscopic methods (1D NMR and ESI-MS) as well as by comparison with the literature data. An antimicrobial assay showed that these aromatic polyketides exhibited no remarkable inhibitory effect on Escherichia coli, Staphyloccocus aureus and Candida albicans. The genomic feature of strain D was analyzed, as well as its biosynthetic gene clusters, using antibiotics and Secondary Metabolite Analysis Shell 5.1.2 (antiSMASH). Plausible biosynthetic pathways for dimeric naphtho-γ-pyrones 3-8 were first proposed in this work. A non-reducing polyketide synthase (PKS) gene D8.t287 responsible for the biosynthesis of these aromatic polyketides 1-8 was identified and characterized by target gene knockout experiment and UPLC-MS analysis.

Five new 2-(2-phenylethyl)chromone derivatives from agarwood.

Agarwood has been used as an incense and in traditional medicines as aphrodisiac, sedative, cardiotonic, and carminative. In this study, five new 2-(2-phenylethyl)chromones (2, 13-16) and eleven known compounds (1, 3-12) were isolated from the agarwood. The structures of the new compounds were determined by 1H-, 13C-, and two-dimensional NMR together with electronic circular dichroism (ECD) spectroscopy. All isolated compounds were evaluated for the phosphodiesterase (PDE) 3A and 5A1 inhibitory activity by the fluorescence polarization method. Dimeric 2-(2-phenylehyl)chromones (13, 14, 16) had potent inhibitory activity to PDE 5A1 with IC50 values of micro molar range (13: 4.2 µM, 14: 7.9 µM, 16: 4.3 µM), whereas they had weak activity to PDE 3A. In contrast, compound (15), which has a phenylpropionic acid moiety instead of the 2-(2-phenylethyl)chromone moiety in the dimers, showed moderate inhibition of both PDE 3A (IC50: 42.6 µM) and PDE 5A1 (IC50: 15.1 µM).

Mycolicibacterium stellerae sp. nov., a rapidly growing scotochromogenic strain isolated from Stellera chamaejasme.

A polyphasic study was undertaken to establish the taxonomic provenance of a rapidly growing Mycolicibacterium strain, CECT 8783T, recovered from the plant Stellera chamaejasme L. in Yunnan Province, China. Phylogenetic analyses based upon 16S rRNA and whole-genome sequences showed that the strain formed a distinct branch within the evolutionary radiation of the genus Mycolicibacterium. The strain was most closely related to Mycolicibacterium moriokaense DSM 44221T with 98.4 % 16S rRNA gene sequence similarity, but was distinguished readily from this taxon by a combination of chemotaxonomic and phenotypic features and by low average nucleotide identity and digital DNA-DNA hybridization values of 79.5 and 21.1 %, respectively. Consequently, the strain is considered, to represent a novel species of Mycolicibacterium for which the name Mycolicibacterium stellerae sp. nov is proposed; the type strain is I10A-01893T (=CECT 8783T=KCTC 19843T=DSM 45590T).

Evaluation of three different artificial agarwood-inducing methods from Aquilaria sinensis using antimicrobial activity.

The aim of this paper was to evaluate the effect of three different approaches for artificially inducing the formation of agarwood over time in young Aquilaria sinensis trees using antimicrobial activity. The antimicrobial activity was determined by a two-fold serial dilution method and minimum inhibitory concentration (MIC) against a panel of microorganisms (two bacterial strains, Staphylococcus aureus and anti-methicillin-resistant Staphylococcus aureus, and seven fungal strains: Penicillium melinii, Penicillium adametzi, Penicillium urticae, Penicillium notatum, Paecilomyces varioti, Mucor saturninus Hagem and Aspergillus niger). The results showed that artificial agarwood obtained by comprehensive stimulated method (formic acid plus fungal inoculation) and extended longer inducing time have better antimicrobial activity, which is similar to the result of chemical analysis. Therefore, it is a beneficial exploration to the first use of antimicrobial activity to evaluate artificial agarwood obtained by different producing methods and different culture time.

Purpurolide A, 5/5/5 Spirocyclic Sesquiterpene Lactone in Nature from the Endophytic Fungus Penicillium purpurogenum.

Purpurolide A (1), an unprecedent sesquiterpene lactone with a rarely encountered 5/5/5 spirocyclic skeleton, along with two new 6/4/5/5 tetracyclic sesquiterpene lactones (2 and 3), were isolated from the cultures of the endophytic fungus Penicillium purpurogenum IMM003. The structures and absolute configurations of 1-3 were established by spectroscopic analysis, single-crystal X-ray diffraction, and calculations of the 13C NMR and ECD data. Compounds 1-3 showed significant inhibitory activity against pancreatic lipase.

New secondary metabolites from the endophytic fungus Fusarium sp. HP-2 isolated from "Qi-Nan" agarwood.

A novel pyrone derivative (1) bearing two fused five-member rings, together with two new naphthalenone derivatives (2, 3), as well as two known compounds (4, 5) were obtained from the endophytic fungus Fusarium sp. HP-2, which was isolated from "Qi-Nan" agarwood. The structures of the new compounds were elucidated by analysis of 1D and 2D NMR, and by HRESIMS spectra, as well as by comparison with the literature. Bioactivity results indicated that compound 3 showed weak acetylcholinesterase inhibitory activity.

Trunk surface agarwood-inducing technique with Rigidoporus vinctus: An efficient novel method for agarwood production.

Only when Aquilaria spp. or Gyrinops spp. trees are wounded, due to insect attack, or microbial invasion, agarwood can be successfully induced. In the present study, a fungus which can induce agarwood formation efficiently was isolated and a suitable method for its application to induce agarwood formation was developed. Rigidoporus vinctus was isolated from the inner layers from infectious A. sinensis trees. When the fermentation liquid of fungi inoculated back to A. sinensis tree, agarwood was found to be induced. In addition, a novel method called trunk surface agarwood-inducing technique (Agar-Sit) was developed to produce agarwood with R. vinctus. The alcohol soluble extract content of the agarwood, up to 38.9%, far higher than the requirement (10%) in Chinese Pharmacopoeia and the six characteristic compounds of agarwood used as Chinese Medicinal Materials were all detected. Their relative percentages of the sesquiterpenes in the essential oil were 22.76%. This is the first report of the Agar-Sit and also the application of R. vinctus in agarwood induction. According to the results, when the combination of Agar-Sit and R. vinctus is used agarwood can be induced with high yield and good quality.

Monitoring the Chemical Profile in Agarwood Formation within One Year and Speculating on the Biosynthesis of 2-(2-Phenylethyl)Chromones.

Agarwood is highly valued for its uses as incense, perfume, and medicine. However, systematic analyses of dynamic changes of secondary metabolites during the process of agarwood formation have not yet been reported. In this study, agarwood was produced by transfusing the agarwood inducer into the trunk of Aquilaria sinensis, and changing patterns of chemical constituents, especially 2-(2-phenylethyl)chromones (PECs), in wood samples collected from the 1st to 12th month, were analyzed by GC-EI-MS and UPLC-ESI-MS/MS methods. Aromatic compounds, steroids, fatty acids/esters, sesquiterpenoids, and PECs were detected by GC-MS, in which PECs were the major constituents. Following this, UPLC-MS was used for further comprehensive analysis of PECs, from which we found that 2-(2-phenylethyl)chromones of flindersia type (FTPECs) were the most abundant, while PECs with epoxidated chromone moiety were detected with limited numbers and relatively low content. Speculation on the formation of major FTPECs was fully elucidated in our context. The key step of FTPECs biosynthesis is possibly catalyzed by type III polyketide synthases (PKSs) which condensate dihydro-cinnamoyl-CoA analogues and malonyl-CoA with 2-hydroxy-benzoyl-CoA to produce 2-(2-phenyethyl)chromone scaffold, or with 2,5-dihydroxybenzoyl-CoA to form FTPECS with 6-hydroxy group, which may serve as precursors for further reactions catalyzed by hydroxylase or O-methyltransferase (OMT) to produce FTPECs with diverse substitution patterns. It is the first report that systematically analyzed dynamic changes of secondary metabolites during the process of agarwood formation and fully discussed the biosynthetic pathway of PECs.

Secondary metabolites from the Colletotrichum gloeosporioides A12, an endophytic fungus derived from Aquilaria sinensis.

Two new cyclohexene derivatives colletotricones A and B (1 and 2) and a new thiazole derivative colletotricole A (5), along with six known natural metabolites were isolated from the extract of Colletotrichum gloeosporioides A12, an endophytic fungus derived from Aquilaria sinensis. Among them, the colletotricones A and B possess a cyclohexenone skeleton, whereas the colletotricole A is a thiazole derivative. Their structures were fully assigned with the aid of extensive spectroscopic analysis and data from the literature. Moreover, cytotoxic activity in vitro of compounds 1 and 3-9 were evaluated against MCF-7, NCI-H460, HepG-2 and SF-268 tumour cell lines. The new compound 1 exhibited growth inhibitory activity against all the four tumour cell lines with IC50 values ranging from 15.7 to 46.8 µM.

Filimonas aquilariae sp. nov., isolated from agarwood chips.

A polyphasic approach was used to characterize a Gram-staining negative bacterium (designated strain CC-YHH650T) isolated from agarwood chips. Strain CC-YHH650T was aerobic and rod-shaped, able to grow at 15-37 °C (optimal 30 °Ð¡), pH 6.0-8.0 (optimal 7.0) and 0-1 % (w/v) NaCl (optimal 0 %). Phylogenetic analysis based on 16S rRNA genes revealed that strain CC-YHH650T shared highest sequence similarities with Filimonas lacunae (97.5 %), F. zeae (97.4 %), F. endophytica (97.3 %) and F. aurantiibacter (93.0 %), and lower sequence similarity with other genera (less than 93.0 %). The levels of DNA-DNA relatedness between strain CC-YTH209T, F. lacunae, F. endophytica and F. zeae were estimated to be 18.3, 6.1, 24.7 % (the reciprocal values were 9.8, 8.8, 18.3 %). The major fatty acids were iso-C15 : 0, iso-C15 : 1 G, C16 : 0 3-OH, iso-C17 : 0 3-OH and C16 : 1ω7c/C16 : 1ω6c. The polar lipid profile contained phosphatidylethanolamine, an unidentified phospholipid, two unidentified aminophospholipids, three unidentified aminolipids and three unidentified lipids. The DNA G+C content was 46.6 mol% and the predominant quinone was menaquinone-7 (MK-7). The major polyamine was sym-homospermidine. Based on the distinct phylogenetic, phenotypic and chemotaxonomic traits together with results of comparative 16S rRNA gene sequence analysis, strain CC-YHH650T is considered to represent a novel species of the genus Filimonas, for which the name Filimonas aquilariae sp. nov. is proposed. The type strain is CC-YHH650T (=BCRC 80935T=JCM 31197T).

Agarwood Formation Induced by Fermentation Liquid of Lasiodiplodia theobromae, the Dominating Fungus in Wounded Wood of Aquilaria sinensis.

Agarwood is broadly used in incense and medicine. Traditionally, agarwood formation is induced by wounding the trunks and branches of some species of Aquilaria spp., including A. sinensis. As recently evidenced, some fungi or their fermentation liquid may have the potential of inducing agarwood formation. The present study aimed to analyze the fungi isolated from an agarwood-producing A. sinensis tree and subsequently identify the fungi capable of promoting agarwood formation. We identified a total of 110 fungi isolates based on their morphological characteristics and rDNA ITS sequences. These isolates came from four different layers (namely the decomposing layer, agarwood layer, transition layer, and normal layer) near the agarwood formation site of the trunk. According to the experimental results, most of them belonged to Dothideomycetes (81.82%), while the others to Sordariomycetes (13.64%) or Eurotiomycetes (4.55%). Of note, 88 isolates were shown belonging to the species of Lasiodiplodia theobromae that are most frequently isolated from different layers. In addition, when the fermentation liquid of two isolates of L. theobromae (AF4 and AF12) and one isolate of Fusarium solani (AF21) was inoculated into the A. sinensis wood using the Agar-Wit technique, promoted agarwood formation was observed; however, the effect of AF21 did not keep stable in the later test, while AF4 and AF12 still functioned 1 year later. This study may lay a foundation for exploring the underlying mechanism of agarwood formation as well as fungi application in agarwood production.

Fungal Endophytes: an Alternative Source for Production of Volatile Compounds from Agarwood Oil of Aquilaria subintegra.

Fungal endophytes are microorganisms that are well-known for producing a diverse array of secondary metabolites. Recent studies have uncovered the bioprospecting potential of several plant endophytic fungi. Here, we demonstrate the presence of highly bioactive fungal endophytic species in Aquilaria subintegra, a fragrant wood plant collected from Thailand. Thirty-three fungal endophytic strains were isolated and further identified to genus level based on morphological characteristics. These genera included Colletotrichum, Pestalotiopsis, Fusarium, Russula, Arthrinium, Diaporthe and Cladosporium. All strains were cultured on potato dextrose broth for 30 days prior to partitioning with ethyl acetate. The volatile compounds of all extracts were investigated by gas chromatography-mass spectrometry (GC-MS). Four strains-Arthrinium sp. MFLUCC16-0042, Colletotrichum sp. MFLUCC16-0047, Colletotrichum sp. MFLUCC16-0048 and Diaporthe sp. MFLUCC16-0051-produced a broad spectrum of volatile compounds, including ß-agarofuran, α-agarofuran, δ-eudesmol, oxo-agarospirol, and ß-dihydro agarofuran. These compounds are especially important, because they greatly resemble those originating from the host-produced agarwood oil. Our findings demonstrate the potential of endophytic fungi to produce bioactive compounds with applications in perfumery and cosmetic industries. Antioxidant activity of all extracts was also evaluated by using 2,2-diphenyl-2-picrylhydrazyl radical scavenging assays. The ethyl acetate extract of Diaporthe sp. MFLUCC16-0051 demonstrated superior antioxidant capacity, which was comparable to that of the gallic acid standard. Our results indicate that the MFLUCC16-0051 strain is a resource of natural antioxidant with potential medicinal applications.