Mangrove endophytic fungi can produce impressive quantities of metabolites with promising antioxidant activities that may be useful to humans as novel physiological agents. In this study, we investigated the phylogenetic diversity and antioxidant potential of 46 fungal endophytes derived from the mangrove species Rhizophora stylosa and R. mucronata from the South China Sea. The fungal isolates were identified using a combination of morphological characteristics and phylogenetic analysis of the internal transcribed spacer (ITS) sequences. Seventeen genera belonging to 8 taxonomic orders of Ascomycota were discovered, specifically, Botryosphaeriales, Capnodiales, Diaporthales, Eurotiales, Glomerellales, Hypocreales, Pleosporales, and Xylariales. The most abundant fungal orders included Xylariales (35.49%) and Diaporthales (27.61%), which were predominantly represented by the culturable species Pestalotiopsis sp. (34.54%) and Diaporthe sp. (18.62%). The stems showed more frequent colonization and species diversity than the roots, leaves, hypocotyls, and flower tissues of the host plant. The antioxidant activities of all the isolated fungal extracts on four different culture media were assessed using improved 2,2′-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonicacid) (ABTS) methods. A relatively high proportion (84.8%) of the isolates displayed antioxidant capacity (%RSA > 50%). Further research also provided the first evidence that HQD-6 could produce flufuran as a significant radical scavenger with IC₅₀ values of 34.85±1.56 and 9.75±0.58 μg/mL, respectively. Our findings suggest that the utilization of a biotope such as that of the endophytic fungal community thriving on the mangrove plants R. stylosa and R. mucronata may be suitable for use as a sustainable resource for natural antioxidants.

Chemical examination of Chinese mangrove Rhizophora mucronata endophytic Pestalotiopsis sp., yielded 11 known metabolites with various structure types, including demethylincisterol A₃ (1), dankasterone B (2), (22E, 24R)-ergosta-7,9(11), 22-triene-3β, 5α, 6α-triol (3), ergosta-5,7,22-trien-3-ol (4), 5, 8-epidioxy-5, 8-ergosta-6, 22E-dien-3-ol (5), stigmastan-3-one (6), stigmast-4-en-3-one (7), stigmast-4-en-6 -ol-3-one (8), flufuran (9), (2-cis, 4-trans)-abscisic acid (10), similanpyrone B (11). Their structures were unambiguously elucidated on the basis of extensive NMR spectroscopic and mass spectrometric analyses. Compounds 1, 4, 6–9 showed significant in vitro cytotoxicity against the human cancer cell lines Hela, A549 and HepG, of which compound 1 was the most potential with IC₅₀ values reaching nM degree ranging from 0.17 to 14.16 nM. Flow cytometric investigation demonstrated that compound 1 mainly inhibited cell cycle at G₀/G₁ phase in a dose-dependent manner with a significant induction of apoptosis on the three tested cell lines. The involvement of the mitochondria in compound 1 induced apoptosis was investigated using MMP. We suggested that R. mucronata endophytic Pestalotiopsis sp. contained a potential anticancer compound demethylincisterol A₃.

Mangrove endophytic fungi are tolerant to numerous stresses and are inevitably capable of exhibiting excellent biological activity by producing impressive numbers of metabolites with special biological functions, based on previous work on the biological potential of mangrove-derived endophytic fungi. To obtain marked antimicrobial and cytotoxic fermentation products of culturable endophytic fungi from mangrove forests, our research evaluated the antimicrobial and cytotoxic activities of crude extracts of endophytic fungi from Rhizophora stylosa and Rhizophora mucronata. Forty-six fungal isolates were cultured on four different media, namely, dextrose agar (PDA), Czapek’s agar (CZA), rice medium (RM) and grain medium (GM) and harvested by ethyl acetate solvent at 40 days. The extracts were tested for antimicrobial activity by the microdilution method against the gram-negative bacteria Pseudomonas adaceae (PA), gram-positive bacteria Enterococcus faecalis (EF), methicillin-resistant Staphylococcus aureus (MRSA) and pathogenic fungus Monilia albicans (MA). The cytotoxic activity of the extracts was evaluated by MTT assay using A549 human lung cancer cells, HeLa human cervical carcinoma cells, and HepG2 human hepatocellular cells. The results showed that rice medium could promote the secretion of antimicrobial and antitumour secondary metabolites of endophytic fungi in comparison with other cultivation media. Seventeen strains (68%) from R. stylosa exhibited inhibitory effects on indicators, especially N. protearum HHL46, which could inhibit the growth of four microbes with MIC values reaching 0.0625 mg/mL. Fifteen strains (71.4%) from R. mucronata displayed activities against human pathogenic microbes; in particular, Pestalotiopsis sp. HQD6 and N. protearum HQD5 could resist the growth of four microbes with MIC values ranging from 0.015 to 1 mg/mL. In the cytotoxicity assay, the extracts of 10 strains (40%), 9 strains (40%) and 13 strains (52%) of R. stylosa and 13 strains (61.9%), 10 strains (47.6%) and 10 strains (47.6%) of R. mucronata displayed cytotoxicity against A549, HeLa and HepG2 cancer cells with cell viability values ≤ 50%. Neopestalotiopsis protearum HHL46, Phomopsis longicolla HHL50, Botryosphaeria fusispora HQD83, Fusarium verticillioides HQD48 and Pestalotiopsis sp. HQD6 displayed significant antitumour activity with IC₅₀ values below 20 μg/mL. These results highlighted the antimicrobial and antitumour potential of endophytic fungi from R. stylosa and R. mucronata and the possibility of exploiting their antimicrobial and cytotoxic agents.

This study was designed to investigate the antioxidant properties of the extracts and subfractions of various polarities from Clerodendrum cyrtophyllum Turcz leaves and the related phenolic compound profiles. The ethyl acetate fraction (EAF) showed the most potent radical-scavenging activity for DPPH radicals, ABTS radicals, and superoxide anion (O₂^·-) radicals as well as the highest reducing power of the fractions tested; the n-butyl alcohol fraction (BAF) was the most effective in scavenging hydroxyl radical (OH^·), and the dichloromethane fraction (DMF) exhibited the highest ferrous ion chelating activity. Twelve phenolic components were identified from the EAF of C. cyrtophyllum. Additionally, acteoside (1) was found to be a major component (0.803 g, 0.54%) and show DPPH and ABTS radical scavenging activities with IC₅₀ values of 79.65±3.4 and 23.00±1.5 μg/ml, indicating it is principally responsible for the significant total antioxidant effect of C. cyrtophyllum. Our work offers a theoretical basis for further utilization of C. cyrtophyllum as a potential source of natural, green antioxidants derived from plants.

Mangrove endophytic fungi were proved to be a prolific resources for bioactive secondary metabolites. Two new polyketides, pestalotiopin B (1) and pestalotiopyrone N (2) were obtained from the ethyl acetate extracts of the rice solid cultures of the mangrove endophytic fungus Pestalotiopsis sp. HQD-6. The structures of 1 and 2 were determined by detailed interpretation of NMR spectroscopic analysis and HR-ESIMS. All of the isolates were evaluated for their antimicrobial, antioxidant and cytotoxic activities. Graphical abstract