Biocontrol efficiency of native Bacillus sp. HC-9 on honeysuckle leaf spot caused by Alternaria alternata

Background: The growing emphasis on sustainable agriculture has intensified interest in plant growth stimulants derived from natural, non-toxic components. Among these innovations is the micronutrient formulation “Complex-Co,” a tartaric acid–based chelate containing vital elements such as iron (Fe), copper (Cu), zinc (Zn), and boron (B). These micronutrients play key roles in plant physiology, including enzyme function, photosynthesis, growth regulation, and stress tolerance. As a plant growth stimulant, “Complex-Co” holds promise for enhancing plant vigor and strengthening resistance against phytopathogenic and postharvest spoilage fungi, major contributors to reduced crop yields and quality deterioration. This study is the first to evaluate the antifungal potential of the multifunctional Complex-Co formulation, which combines copper, zinc, and iron with a biogenic nitrogen-containing tartaric acid derivative. Its unique composition enables simultaneous stimulation of plant growth and inhibition of key postharvest phytopathogens, highlighting its potential for sustainable crop protection and functional-food applications. Objective: This study aims to investigate the antifungal efficacy of the micronutrient-based plant growth promoter “Complex-Co” against selected phytopathogenic and postharvest spoilage fungi. The research evaluates explicitly the antifungal activity of the formulation at concentrations commonly used in standard plant nutrition practices. Furthermore, the study compares the effectiveness of “Complex-Co” with that of conventional copper-based antifungal treatments. Materials and Methods: To assess antifungal activity, fungal strains from the genera Alternaria, Aspergillus, Cladosporium, Rhizomucor, and Rhizopus were used as test organisms. Cultures were grown on potato dextrose agar (PDA) in 90 mm × 15 mm Petri dishes and incubated at 25 ± 1 °C. Once the fungal mycelia reached a diameter of 4-5cm, four sterile filter paper disks (6 mm in diameter) were placed approximately 5mm from the edge of the colony. The “Complex-Co” preparation was tested at 10-fold and 100-fold dilutions, alongside a similarly diluted copper-based antifungal formulation for comparison. Each disk was treated with 20 µL of the respective solution. Following treatment, the plates were incubated at 25°C, and fungal growth inhibition was monitored for 2 to 7 days. Results: The antifungal activity of “Complex-Co” was evaluated against five phytopathogenic fungi: Alternaria alternata, Aspergillus flavus, Cladosporium herbarum, Rhizomucor miehei, and Rhizopus oryzae. At a 1:100 dilution, which corresponds to a Cu²⁺ concentration of 0.125 g/L, inhibition was limited, with zones ≤ 8 mm. At a 1:10 dilution ([Cu²⁺]=1.25 g/L), antifungal activity increased markedly, particularly against Alternaria alternata (13 mm), Aspergillus flavus (15 mm), and Cladosporium herbarum (17 mm). Compared with the commercial fungicide “Avgust Ordan” at equivalent copper concentrations, “Complex-Co” showed comparable or superior inhibition against certain fungi. UV-Visible spectroscopy confirmed the presence of proteinaceous and amino acid components, suggesting their roles in copper chelation, solubility, stabilization, and enhanced antifungal activity. These results demonstrate a concentration-dependent antifungal activity and highlight the potential of “Complex-Co” as a multifunctional plant protection and nutrient formulation. Conclusion: Micronutrient “Complex-Co” is demonstrating its potential as a broad-spectrum, environmentally friendly alternative to chemical antifungal treatments. The formulation, which includes essential micronutrients, not only promotes plant growth but also enhances resistance to fungal pathogens. This can reduce reliance on synthetic pesticides, improve crop quality, and increase food security, offering a sustainable solution for both agricultural productivity and public health. Keywords: Chelated Micronutrient, Natural Tartaric Acid, Antifungal Activity, Sustainable Agriculture, Postharvest spoilage fungi.

Microbial interactions, which regulate the dynamics of eco- and agrosystems, can be harnessed to enhance antagonism against phytopathogenic fungi in agriculture. This study tests the hypothesis that plant growth-promoting rhizobacteria (PGPR) can also be potential biological control agents (BCAs). Antifungal activity assays against potentially phytopathogenic fungi were caried out using cultures and cell-free filtrates of nine PGPR strains previously isolated from agricultural soils. Cultures of Bacillus sp. BS36 inhibited the growth of Alternaria sp. AF12 and Fusarium sp. AF68 by 74 and 65%, respectively. Cell-free filtrates of the same strain also inhibited the growth of both fungi by 54 and 14%, respectively. Furthermore, the co-cultivation of Bacillus sp. BS36 with Pseudomonas sp. BS95 and the target fungi improved their antifungal activity. A subsequent metabolomic analysis using Fourier-transform ion cyclotron resonance mass spectrometry (FTICR-MS) identified fengycin- and surfactin-like lipopeptides (LPs) in the Bacillus sp. BS36 cell-free filtrates, which could explain their antifungal activity. The co-production of multiple families of LPs by Bacillus sp. BS36 is an interesting feature with potential practical applications. These results highlight the potential of the PGPR strain Bacillus sp. BS36 to work as a BCA and the need for more integrative approaches to develop biocontrol tools more accessible and adoptable by farmers.

BACKGROUNDEssential oils (EOs) extracted from eight culinary herbs and spices, some endemic to Macaronesia flora (Cedronella canariensis, Clinopodium ascendens and Laurus novocanariensis) and others common in Mediterranean cuisine (Cinnamomum burmannii, Ocimum gratissimum, Origanum vulgare subsp. virens, Syzygium aromaticum and Thymus vulgaris) were evaluated via direct‐contact bioassays, both in vitro and in vivo, against three species of phytopathogenic fungi (Alternaria alternata, Botrytis cinerea, and Fusarium oxysporum) and the obligatory biotrophic fungus Oidium farinosum. Carvacrol‐rich Origanum vulgare was selected to study the influence of isomerism on bioactivity, because Thymus vulgaris is rich in thymol, a structural isomer of carvacrol.RESULTSFive EOs exhibited strong to moderate antifungal activity and were further screened at lower concentrations to assess their toxicity and growth inhibition thresholds. Cedronella canariensis showed no antifungal activity, whereas Laurus novocanariensis demonstrated only weak activity against Botrytis cinerea. This fungus was more susceptible to the tested EO than the other phytopathogenic fungi. Origanum vulgare was slightly more effective than Thymus vulgaris, for all three phytopathogenic fungi. Ocimum gratissimum, Syzygium aromaticum, and Cinnamomum burmannii significantly reduced powdery mildew severity, whereas Origanum vulgare and Thymus vulgaris showed moderate activity.CONCLUSIONPhenylpropanoid‐rich EOs, such as Cinnamomum burmannii, Ocimum gratissimum, and S. aromaticum, exhibited the strongest antifungal activity in both bioassays with phytopathogenic and obligate biotrophic fungi. Our results confirm the higher antifungal activity of the phenylpropanoid class of EOs. © 2025 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Plant roots are associated with diverse communities of endophytic bacteria which do not exert adverse effects. The diversity of bacterial endophytes associated with ginseng roots cultivated in three different areas in Korea was investigated. Sixty-three colonies were isolated from the interior of ginseng roots. Phylogenetic analysis based on 16S rRNA gene sequences showed that the isolates belonged to three major phylogenetic groups: the high G+C Gram-positive bacteria (HGCGPB), low G+C Gram-positive bacteria (LGCGPB), and the Proteobacteria. The dominant species at the three different ginseng growing areas were: HGCGPB at Ganghwa (55.0%), LGCGPB at Geumsan (45.5%), and Proteobacteria at Jinan (61.9%). Most cellulase-, xylanase-, and pectinase-producing colonies among the isolates belong to the LGCGPB group, except for Pectobacterium carotovora which belonged to the Proteobacteria. The 13 isolates belonging to LGCGPB and Proteobacteria were assessed for their antifungal activity against phytopathogenic fungi such as Rhizoctonia solani. Among them, Paenibacillus polymyxa GS01, Bacillus sp. GS07, and Pseudomonas poae JA01 show potential activity as biocontrol agents against phytopathogenic fungi. Finally, most of the low G+C Gram-positive bacteria with antifungal activity against phytopathogenic microorganisms showed cellulolytic enzyme activity while some Proteobacteria with the antifungal activity and the high G+C Gram-positive bacteria did not show any cellulolytic activity.

Honey is thought to act against microbes and regulates microbiota balance, and this is mainly attributed to the enzymatic production of hydrogen peroxide, high osmolarity, and nonperoxidase factors, for example, lysozyme and botanical sources of nectar, while the effect of honey's probiotic is recently considered. The study of honey as source of beneficial microbes is understudied. The purpose of this study was to screen for the beneficial microorganisms in honey with antagonistic property against important pathogens and the mechanism of antimicrobial activity and thus play a beneficial role as probiotics. The results showed that one out of the fourteen bacterial isolates had antimicrobial activity and was identified as Bacillus Sp. A2 by 16S rRNA sequence and morphology. Antimicrobial activity of the isolate against C. albicans, E. coli, and S. aureus was confirmed by Agar well diffusion and liquid coculture assays, and the propagation of those microbes was significantly inhibited after treatment with the isolate Bacillus sp. A2 (p < .05) in comparison with untreated negative control and positive control (fluconazole, chloramphenicol, L. plantarum). The morphological changes including the distorted shape with indentations and leakages (SEM), damaged cell membrane, and cell wall with the disintegration and attachment of the Bacillus sp. A2 (TEM) in treated C. albicans were observed. Meanwhile, reactive oxygen species accumulation and decreased mitochondrial membrane potential were detected in treated C. albicans. These results revealed that the isolate Bacillus sp. A2 from honey has significant antimicrobial activity (p < .05) against C. albicans in comparison with untreated negative control and positive control L. plantarum, which depends on the accumulation of reactive oxygen species, mitochondrial damage, and the cell apoptosis. We concluded that the Bacillus sp. A2 possess the antimicrobial property, which may contribute to regulation of host's microbiota as a beneficial microbe or probiotic.

New bacterial strains of Bacillus licheniformis and sp were isolated from the rhizosphere of potato, in front of the sea (Bejaia, Algeria). Strains were identified by phenotypic analyses: Gram s taining, mobility, catalase, oxidase, form of colony and cell morphology. Metabolic characters such carbon source, sugars, organic acids, aerobic/anaerobic respiratory system are revealed by API system 50CHB/E. The 16SrRNA gene was isolated and sequenced (1500 bp), and the phylogenic tree was established (bootstrap method). The first strain (RBA-E23) showed a very high similarity to Bacillu s licheniformis (DQ082997.1) (98%) and B. licheniformis (AF372616.1), but the second (RBA-E32) is classified as the out-gro up. Strains are halotolerants 300 and 400 mM/l NaCl, respectively. They used glycine betaine and proline as synthetic osmoprotectants, and marine algal extracts of Ulva lactucaas natural osmoprotectant. The two strains produce: high amount of auxine-phytohormone (IAA: 78 and 101 mg/l); different enzymes such as: cellulase, chitinase, protease, lipase, amylase and urease; siderophores and solubilize ino rganic phosphorus. The study of antifungal strains activity toward nine phyto pathogenic fungi Verticillium dahlia, Fusarium oxysporum, Botryotinia fuckiliana, Phytophthor a cinamomi, Phytophthora cactorum, Colletotrichum acutatum, Botrytis cinerea, Aspergillus Niger and Aspergillus flavus, shows an interesting inhibition activity with variable percentage of Growth inhibition (PGI %) against phytopathogens. Both of bacteria displayed more benefic characters, while the enzymes can be involved in the organic matter decomposition; utilization of the phytohormone-IAA as simulator of vegetable growth. Siderophores and phosphate solubilization are interesting in the fe rtilization of soil. Finally, strains could have potential application against phytopathogenic fungi.

We evaluated the antagonistic activity of 23 strains of Bacillus spр. against phytopathogenic fungi Fusarium oxysporum, F. culmorum, F. moniliforme, Cladosporium herbarum, Alternaria alternata and Aspergillus niger. The antagonistic activity was tested by agar diffusion (the method of blocks). For determining the influence of bacteria on barley plants , ardent seeds were treated by cultural liquid (dilution 1 : 10) for 2 hours and germinated in Petri dishes on moist filter paper. The fungistatic effect of Bacillus sрp. separately and in combination with entomopathogens (in equal ratio) was determined by the level of inhibition of the fungi Fusarium spp. on a solid nutrient medium with 5% of the culture liquid. Insecticidal activity of microorganisms was determined in the model experiments by the percentage of death of the caterpillar Archips podana Scop. Strains of Bacillus sp. KMB-3 and Bacillus sp. KMB-6 inhibited the growth of all test cultures (zones of growth inhibition 11.4–30.6 and 11.5–29.4 mm, respectively). We established the absence of antagonism between selected strains and entomopathogenic bacteria Bacillus thuringiensis IMB-7186, fungi Beauveria bassiana IMB-F-100043. We found that treatment of barley seeds with culture liquids of Bacillus sp. KMB-3 and Bacillus sp. KMB-6 didn’t have a negative effect on the morphometric indices and dry weight of seedlings. We established that the highest percentage of growth inhibition of F. culmorum IMB-F-50716 was provided by a complex of Bacillus sp. KMB-3, B. bassiana IMB-F-100043 and B. thuringiensis IMB-7186, whose action was at the same level as the action of monoculture Bacillus sp. KMB-3 (85.4% and 84.7%, respectively). The highest percentage inhibition of growth of F. oxysporum ІМВ-F-54201 was provided by a complex of strains of Bacillus sp. KMB-3 and B. bassiana IMB-F-100043, whose effect was slightly inferior to that of the monoculture Bacillus sp. KMB-3 (68.4% and 75.1%, respectively). The insecticidal activity of complexes Bacillus sp. KMB-3, B. bassiana IMB-F-100043, B. thuringiensis IMB-7186 or Bacillus sp. KMB-6, B. bassiana IMB-F-100043, B. thuringiensis IMB-7186 insignificantly differed from that of the complex entomopathogens B. bassiana IMB-F-100043 and B. thuringiensis IMB-7186 (71.1%, 73.3% death versus 80.0%). The selected microbial complexes can be considered as promising for the development of a preparation for the protection of plants against fungal diseases and harmful insects.

Endophytes are potential source of various novel compounds that help to promote plant growth, eliminate plant pathogens, and enable the plant to resist stress-like conditions. The present study aimed at selecting stress-tolerant bacterial endophytes with plant growth-promoting ability from Adhatoda vasica. Salt-tolerant bacterial endophytes were isolated on nutrient agar with 2.5% NaCl from the leaves of Adhatoda vasica collected from Manipur, India. The isolates were screened for stress tolerance, plant growth-promoting traits, antagonism against fungal phytopathogens and plant growth promotion. Sixteen morphologically distinct salt-tolerant bacterial endophytes were isolated from Adhatoda vasica. All bacterial endophytes showed auxin production while phosphate solubilization was shown by 81% isolates, siderophore production by 75%, ACC deaminase by 43.8% isolates, HCN by 50% isolates, and ammonia by 62.5% isolates. Four bacterial isolates showed antagonistic activity against all the test fungal phytopathogens Fusarium verticillioides (MTCC 3322), Curvularia lunata (MTCC 283), and Alternaria alternata (MTCC 1362). Dendrogram generated based on stress tolerance of the bacterial isolates against salinity, temperature, pH, and calcium salts showed 3 clusters and two independent branches. Two bacterial isolates identified based on phenotypic features and 16S rRNA gene sequencing as Bacillus thuringiensis A1B3 and Bacillus sp. A1B6 significantly increased the growth parameters of pea and maize in comparison to uninoculated control in pots under natural conditions. The attributes of stress tolerance, antagonism against fungal pathogens, and plant growth promotion indicated the potential of Bacillus thuringiensis A1B3 and Bacillus sp. A1B6 to be used as microbial inoculant in agriculture under stressed environment.

The aim of the present study was to focus on the growth promotion and wilt diseases protection of tomato plants by Bacillus sp. LBF-01.The plant growth promotion (both in vitro and in vivo) and diseases protection mechanism were studied following standard protocols. This bacterial strain showed positive PGP and antifungal traits against the wilt pathogen Fusarium sp. The seed bacterization with Bacillus sp. LBF-01 improved the germination rate of tomato seeds by 20% compared to the control whereas the vigor index was increased by 854.026%.The percentage of disease protection by Bacillus sp. LBF-01 against root-rot disease inoculated was inversely proportional to the doses of fungal spores inoculum. Under in vivo condition, the PGP bacteria enhanced the seedling growth in nursery as well as field condition. Application of Bacillus sp. LBF-01 enhanced the activity of defense related enzymes in root and leaves. Bacillus sp. LBF-01 treatment had significantly enhanced total chlorophyll and carotenoids contents of leaves by 77.78 and 52.5% over untreated control plants. It is evident from the present study that the application of the Bacillus sp. LBF-01 bacterial strain in tomato soil improved the potentiality of the plant by systemic resistance and control the wilt diseases for tomato production.

As a continuous flow investigation of novel pesticides from natural quinolizidine alkaloids, the chemical compositions of the seeds of Sophora alopecuroides were thoroughly researched. Fifteen new aloperine-type alkaloids (1-15) as well as six known aloperine-type alkaloids (16-21) were obtained from the extract of S. alopecuroides. The structures of 1-21 were confirmed via HRESIMS, NMR, UV, IR, ECD calculations, and X-ray diffraction. The antiviral activities of 1-21 against tobacco mosaic virus (TMV) were detected following the improved method of half-leaf. Compared with ningnanmycin (protective: 69.7% and curative: 64.3%), 15 exhibited excellent protective (71.7%) and curative (64.6%) activities against TMV. Further biological studies illustrated that 15 significantly inhibited the transcription of the TMV-CP gene and increased the activities of polyphenol oxidase (PPO), peroxidase (POD), superoxide dismutase (SOD), and phenylalanine ammonia-lyase (PAL). The antifungal activities of 1-21 against Phytophythora capsica, Botrytis cinerea, Alternaria alternata, and Gibberella zeae were screened according to a mycelial inhibition test. Compound 13 displayed excellent antifungal activity against B. cinerea (EC50: 7.38 μg/mL). Moreover, in vitro antifungal mechanism studies displayed that 13 causes accumulation of reactive oxygen species and finally leads to mycelia cell membrane damage and cell death in vitro.

This study was carried out in order to develop a biological control of anthracnose of red pepper caused by fungal pathogens. In particular, this study focuses on the Colletotrichum species, which includes important fungal pathogens causing a great deal of damage to red pepper. Antagonistic bacteria were isolated from the soil of pepper fields, which were then tested for biocontrol activity against the Colletotrichum gloeosporioides anthracnose pathogen of pepper. Based on the 16S rRNA sequence analysis, the isolated bacterial strain CS-52 was identical to Bacillus sp. The culture broth of Bacillus sp. CS-52 had antifungal activity toward the hyphae and spores of C. gloeosporioides. Moreover, the substances with antifungal activity were optimized when Bacillus sp. CS-52 was grown aerobically in a medium composed of 0.5% glucose, 0.7% K2HPO4, 0.2% KH2PO4, 0.3% NH4NO3, 0.01% MnSO4·7H2O, and 0.15% yeast extract at 30℃. The inhibition of spore formation resulting from cellulase, siderophores, and indole-3-acetic acid (IAA), were produced at 24 h, 48 h, and 72 h, respectively. Bacillus sp. CS-52 also exhibited its potent fungicidal activity against anthracnose in an in vivo test, at a level of 70% when compared to chemical fungicides. These results identified substances with antifungal activity produced by Bacillus sp. CS-52 for the biological control of major plant pathogens in red pepper. Further studies will investigate the synergistic effect promoting better growth and antifungal activity by the formulation of substances with antifungal activity.

Bacillus sp. BSp.3/aM, a beneficial rhizobacteria, was analyzed for the ability to improve plant health of chili by suppressing anthracnose disease. In the dual culture assay, the bacterium Bacillus sp. BSp.3/aM was found inhibitory to Colletotrichum capsica (6 mm). Further, upon seed priming, it reduced the seed-borne incidence of C. capsici (2%) and improved seedling vigor (1374 ± 7.15 vigor index) and germination (98 ± 0.57 %) of chili seedlings. Under greenhouse conditions, seed priming resulted in reducing the anthracnose disease incidence up to 20%. Induction of resistance against invading pathogen is through enhancing the activities of defense-related enzymes and higher accumulation of phenolic compounds in the host plant. The activity of phenylalanine ammonia-lyase (PAL; 95 units) was more at 48 hpi; peroxidase (POX; 6.49 units) at 24 hpi; polyphenol oxidase (PPO; 5.81 units) at 24 hpi and lipoxygenase (LOX; 9.9units) at 24 hpi. Maximum accumulation of the phenolics and chitinase accumulation was observed in BSp.3/aM + pathogen treated seedlings 120 hpi (94.7 μg/g tissue) and at 96 hpi (9.36 units), respectively. Thus, increased activities of defense-related enzymes (PAL, POX, PPO, LOX, and chitinase) correlated well with the decreased anthracnose incidence. Induced systemic resistance (ISR) mediated by PGPR was due to the upregulation of defense-related enzymes and by the accumulation of phenolic compounds.

The abuse of chemical fungicides not only leads to toxic residues and resistance in plant pathogenic fungi, but also causes environmental pollution and side effects on in humans and animals. Based on the antifungal activities of berberine, seven different types of berberine derivatives (A1-G1) were synthesized, and their antifungal activities against six plant pathogenic fungi were evaluated (Rhizoctonia solani, Botrytis cinerea, Fusarium graminearum, Phytophthora capsici, Sclerotinia sclerotiorum, and Magnaporthe oryzae). The results for antifungal activities in vitro showed that berberine derivative E1 displayed good antifungal activity against R. solani with a median effective concentration (EC50 ) of 1.77 μg ml-1 , and berberine derivatives F1 and G1 demonstrated broad-spectrum antifungal activities with EC50 values ranging from 4.43 to 42.23 μg ml-1 against six plant pathogenic fungi. Berberine derivatives (E2-E29, F2-F18, and G2-G9) were further synthesized to investigate the structure-activity relationship (SAR), and compound E20 displayed significant antifungal activity against R. solani with an EC50 value of 0.065 μg ml-1 . Preliminary mechanism studies showed that E20 could cause mycelial shrinkage, cell membrane damage, mitochondrial abnormalities and the accumulation of harmful reactive oxygen species, resulting in cell death in R. solani. Moreover, in vivo experimental results showed that the protective effect of E20 was 97.31% at 5μg ml-1 , which was better than that of the positive control thifluzamide (50.13% at 5μg ml-1 ). Berberine derivative E20 merits further development as a new drug candidate with selective and excellent antifungal activity against R. solani. © 2022 Society of Chemical Industry.

The gummy stem blight caused by Didymella bryoniae (Auersw.) is a major and destructive disease of watermelon in Vietnam. Bacillus species have been used to control plant diseases as biological control agents. This study was carried out at the Department of Crop Production and Plant Protection, Phu Yen province, Vietnam during 2020 – 2021 to determine fungal inhibition of D. bryoniae DB-03 in in vitro and disease suppression of gummy stem blight under greenhouse conditions by Bacillus sp. strains namely S1A1, S1F3, S13E3, S18F11, and S20D12, which were isolated from stem-base of groundnut in central Vietnam. Results revealed that, all the tested strains of Bacillus sp. showed good inhibitory effect on the hyphal growth of D. bryoniae strain DB-03 in potato dextrose broth. Among the strains, Bacillus sp. S20F12 had highest inhibition of the mycelia growth of D. bryoniae strain DB-03 with an antagonistic efficiency of 60.7% at 10 days after fungal inoculation. Bacillus sp. reduced gummy stem blight under greenhouse conditions. At 21 days after fungal inoculation, disease incidences were low in the plants treated with Bacillus sp. S18F11 and S20D12 by 60.0% and 63.3%, respectively. The results would contribute to the knowledge of antagonistic activities of the Bacillus to optimize the biological control programme against D. bryoniae.

Galangin, a flavonoid extracted from the root of the Alpinia officinarum Hence, has been shown to have anticancer properties against several types of cancer cells. However, the influence of galangin on human renal cancer cells remains to be elucidated. In the present study, proliferation of 786-0 and Caki-1 cells was suppressed following exposure to various doses of galangin. Cell invasion and wound healing assays were used to observe the effect of galangin on invasion and migration. The results demonstrated that Galangin inhibited cell invasion by suppressing the epithelial mesenchymal transition (EMT), with an increase in the expression of E-cadherin and decreased expression levels of N-cadherin and vimentin. The apoptosis induced by galangin was analyzed by flow cytometry. The results revealed that galangin induced apoptosis in a dose-dependent manner. The accumulation of reactive oxygen species (ROS) is an important contributing factor for the apoptosis of various types of cancer cell. The dichlorofluorescein-diacetate method was used to determine the level of ROS. Galangin induced the accumulation of intracellular ROS and malondialdehyde, and decreased the activities of total antioxidant and superoxide dismutase in renal cell carcinoma cells. Galangin exerted an antiproliferative effect and inhibited renal cell carcinoma invasion by suppressing the EMT. This treatment also induced apoptosis, accompanied by the production of ROS. Therefore, the present data suggested that galangin may have beneficial effects by preventing renal cell carcinoma growth, inhibiting cell invasion via the EMT and inducing cell apoptosis.