{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,8]],"date-time":"2026-10-08T08:14:26Z","timestamp":1791447266134,"version":"4.3.3"},"reference-count":42,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2024,10,30]],"date-time":"2024-10-30T00:00:00Z","timestamp":1730246400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"KTO Karatay University Technology Transfer Office, R&amp;D Infrastructure"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["JoF"],"abstract":"<jats:p>Candida species, opportunistic pathogens that cause various infections, pose a significant threat due to their ability to form biofilms that resist antifungal treatments and immune responses. The increasing resistance of Candida spp. and the limited availability of effective treatments have prompted the research of natural compounds as alternative therapies. This study assessed the antifungal properties of RA against Candida species, focusing on its impact on C. albicans biofilms and the underlying mechanisms. The antifungal efficacy of RA was evaluated using the CLSI M27-A3 microdilution method on both fluconazole-susceptible and -resistant strains. Biofilm formation by C. albicans was assessed through a crystal violet assay, while its antibiofilm activity was analyzed using an MTT assay and field emission scanning electron microscopy (FESEM). Gene expression related to biofilm formation was studied using quantitative real-time PCR (qRT-PCR), and statistical analysis was performed with an ANOVA. Among the 28 Candida strains tested, RA exhibited minimum inhibitory concentration (MIC) values ranging from 160 to 1280 \u03bcg\/mL. At a 640 \u03bcg\/mL concentration, it significantly reduced the expression of genes associated with adhesion (ALS3, HWP1, and ECE1), hyphal development (UME6 and HGC1), and hyphal cAMP-dependent protein kinase regulators (CYR1, RAS1, and EFG1) in RAS1-cAMP-EFG1 pathway (p &lt; 0.05). FESEM analysis revealed a reduction in hyphal networks and disruptions on the cell surface. Our study is the first to demonstrate the effects of RA on C. albicans adhesion, hyphae development, and biofilm formation through gene expression analysis with findings supported by FESEM. This approach distinguishes our study from previous studies on the effect of RA on Candida. However, the high MIC values of RA limit its antifungal potential. Therefore, more extensive research using innovative methods is required to increase the antifungal effect of RA.<\/jats:p>","DOI":"10.3390\/jof10110751","type":"journal-article","created":{"date-parts":[[2024,10,30]],"date-time":"2024-10-30T09:11:24Z","timestamp":1730279484000},"page":"751","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["Rosmarinic Acid Exhibits Antifungal and Antibiofilm Activities Against Candida albicans: Insights into Gene Expression and Morphological Changes"],"prefix":"10.3390","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1522-6083","authenticated-orcid":false,"given":"Merve","family":"Aydin","sequence":"first","affiliation":[{"name":"Department of Medical Microbiology, Faculty of Medicine, KTO Karatay University, Konya 42020, Turkey"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nurhan","family":"Unusan","sequence":"additional","affiliation":[{"name":"Department of Nutrition and Dietetics, Faculty of Health Sciences, KTO Karatay University, Konya 42020, Turkey"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5004-5958","authenticated-orcid":false,"given":"Esra","family":"Sumlu","sequence":"additional","affiliation":[{"name":"Department of Medical Pharmacology, Faculty of Medicine, KTO Karatay University, Konya 42020, Turkey"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emine Nedime","family":"Korucu","sequence":"additional","affiliation":[{"name":"Department of Molecular Biology and Genetics, Faculty of Science, Necmettin Erbakan University, Konya 42090, Turkey"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,10,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Li, H., Miao, M.X., Jia, C.L., Cao, Y.B., Yan, T.H., Jiang, Y.Y., and Yang, F. (2022). Interactions between Candida albicans and the resident microbiota. Front. Microbiol., 13.","DOI":"10.3389\/fmicb.2022.930495"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1797","DOI":"10.1007\/s10096-020-03912-w","article-title":"Mechanism of Candida pathogenesis: Revisiting the vital drivers","volume":"39","author":"Mba","year":"2020","journal-title":"Eur. J. Clin. Microbiol. Infect. Dis."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Barantsevich, N., and Barantsevich, E. (2022). Diagnosis and Treatment of Invasive Candidiasis. Antibiotics, 11.","DOI":"10.3390\/antibiotics11060718"},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Salazar, S.B., Sim\u00f5es, R.S., Pedro, N.A., Pinheiro, M.J., Carvalho, M.F.N.N., and Mira, N.P. (2020). An Overview on Conventional and Non-Conventional Therapeutic Approaches for the Treatment of Candidiasis and Underlying Resistance Mechanisms in Clinical Strains. J. Fungi, 6.","DOI":"10.3390\/jof6010023"},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Hern\u00e1ndez-Pab\u00f3n, J.C., Tabares, B., Gil, \u00d3., Lugo-S\u00e1nchez, C., Santana, A., Bar\u00f3n, A., and Firacative, C. (2024). Candida Non-albicans and Non-auris Causing Invasive Candidiasis in a Fourth-Level Hospital in Colombia: Epidemiology, Antifungal Susceptibility, and Genetic Diversity. J. Fungi, 10.","DOI":"10.3390\/jof10050326"},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Whaley, S.G., Berkow, E.L., Rybak, J.M., Nishimoto, A.T., Barker, K.S., and Rogers, P.D. (2017). Azole Antifungal Resistance in Candida albicans and Emerging Non-albicans Candida Species. Front. Microbiol., 7.","DOI":"10.3389\/fmicb.2016.02173"},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Kitaya, S., Kanamori, H., Katori, Y., and Tokuda, K. (2023). Clinical Features and Outcomes of Persistent Candidemia Caused by Candida albicans versus Non-albicans Candida Species: A Focus on Antifungal Resistance and Follow-Up Blood Cultures. Microorganisms, 11.","DOI":"10.3390\/microorganisms11040928"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Arastehfar, A., Gabald\u00f3n, T., Garcia-Rubio, R., Jenks, J.D., Hoenigl, M., Salzer, H.J.F., Ilkit, M., Lass-Fl\u00f6rl, C., and Perlin, D.S. (2020). Drug-Resistant Fungi: An Emerging Challenge Threatening Our Limited Antifungal Armamentarium. Antibiotics, 9.","DOI":"10.3390\/antibiotics9120877"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"2527","DOI":"10.2174\/1568026619666191025152412","article-title":"New and Promising Chemotherapeutics for Emerging Infections Involving Drug-resistant Non-albicans Candida Species","volume":"19","author":"Silva","year":"2019","journal-title":"Curr. Top. Med. Chem."},{"key":"ref_10","doi-asserted-by":"crossref","unstructured":"Chow, E.W.L., Pang, L.M., and Wang, Y. (2021). From Jekyll to Hyde: The Yeast-Hyphal Transition of Candida albicans. Pathogens, 10.","DOI":"10.20944\/preprints202106.0386.v1"},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Watchaputi, K., Jayasekara, L.A.C.B., Ratanakhanokchai, K., and Soontorngun, N. (2023). Inhibition of cell cycle-dependent hyphal and biofilm formation by a novel cytochalasin 19,20-epoxycytochalasin Q in Candida albicans. Sci. Rep., 13.","DOI":"10.1038\/s41598-023-36191-4"},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Poon, Y., and Hui, M. (2023). Inhibitory effect of lactobacilli supernatants on biofilm and filamentation of Candida albicans, Candida tropicalis, and Candida parapsilosis. Front. Microbiol., 14.","DOI":"10.3389\/fmicb.2023.1105949"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Malinovsk\u00e1, Z., \u010conkov\u00e1, E., and V\u00e1czi, P. (2023). Biofilm Formation in Medically Important Candida Species. J. Fungi, 9.","DOI":"10.3390\/jof9100955"},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Kaur, J., and Nobile, C.J. (2023). Antifungal drug-resistance mechanisms in Candida biofilms. Curr. Opin. Microbiol., 71.","DOI":"10.1016\/j.mib.2022.102237"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Czajka, K.M., Venkataraman, K., Brabant-Kirwan, D., Santi, S.A., Verschoor, C., Appanna, V.D., Singh, R., Saunders, D.P., and Tharmalingam, S. (2023). Molecular Mechanisms Associated with Antifungal Resistance in Pathogenic Candida Species. Cells, 12.","DOI":"10.3390\/cells12222655"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"299","DOI":"10.1080\/14787210.2017.1268530","article-title":"Future therapies targeted towards eliminating Candida biofilms and associated infections","volume":"15","author":"Bandara","year":"2017","journal-title":"Expert. Rev. Anti-Infect. Ther."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"e01719-20","DOI":"10.1128\/AAC.01719-20","article-title":"The Future of Antifungal Drug Therapy: Novel Compounds and Targets","volume":"65","author":"Dasilva","year":"2021","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"557","DOI":"10.1038\/s41579-022-00720-1","article-title":"Tackling the emerging threat of antifungal resistance to human health","volume":"20","author":"Fisher","year":"2022","journal-title":"Nat. Rev. Microbiol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"399","DOI":"10.1007\/s13237-022-00405-3","article-title":"Plant-derived natural products for drug discovery: Current approaches and prospects","volume":"65","author":"Nasim","year":"2022","journal-title":"Nucleus"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Guan, H., Luo, W., Bao, B., Cao, Y., Cheng, F., Yu, S., Fan, Q., Zhang, L., Wu, Q., and Shan, M. (2022). A Comprehensive Review of Rosmarinic Acid: From Phytochemistry to Pharmacology and Its New Insight. Molecules, 27.","DOI":"10.3390\/molecules27103292"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Nadeem, M., Imran, M., Gondal, T.A., Imran, A., Shahbaz, M., Amir, R.M., Sajid, M.W., Qaisrani, T.B., Atif, M., and Hussain, G. (2019). Therapeutic potential of rosmarinic acid: A comprehensive review. Appl. Sci., 9.","DOI":"10.3390\/app9153139"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"182","DOI":"10.1016\/j.tifs.2021.03.015","article-title":"Rosmarinic acid-From bench to valuable applications in food industry","volume":"117","author":"Marchev","year":"2021","journal-title":"Trends Food Sci. Technol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"205","DOI":"10.1007\/s12272-022-01378-2","article-title":"Biomedical features and therapeutic potential of rosmarinic acid","volume":"45","author":"Noor","year":"2022","journal-title":"Arch. Pharm. Res."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"358","DOI":"10.5455\/jice.20160906035020","article-title":"Antibacterial synergy between rosmarinic acid and antibiotics against methicillin-resistant Staphylococcus aureus","volume":"5","author":"Ekambaram","year":"2016","journal-title":"J. Intercult. Ethnopharmacol."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Kernou, O.N., Azzouz, Z., Madani, K., and Rijo, P. (2023). Application of Rosmarinic Acid with Its Derivatives in the Treatment of Microbial Pathogens. Molecules, 28.","DOI":"10.3390\/molecules28104243"},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Cheah, H.L., Lim, V., and Sandai, D. (2014). Inhibitors of the glyoxylate cycle enzyme ICL1 in Candida albicans for potential use as antifungal agents. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0095951"},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Bittner Fialov\u00e1, S., Kello, M., \u010coma, M., Slobodn\u00edkov\u00e1, L., Drobn\u00e1, E., Holkov\u00e1, I., Garajov\u00e1, M., Mrva, M., Zachar, V., and Luk\u00e1\u010d, M. (2019). Derivatization of Rosmarinic Acid Enhances its in vitro Antitumor, Antimicrobial and Antiprotozoal Properties. Molecules, 24.","DOI":"10.3390\/molecules24061078"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"521","DOI":"10.1016\/j.sajb.2021.11.050","article-title":"Rosmarinic acid\u2013Modes of antimicrobial and antibiofilm activities of common plant polyphenol","volume":"146","author":"Ivanov","year":"2022","journal-title":"S. Afr. J. Bot."},{"key":"ref_29","unstructured":"Clinical Laboratory Standards Institute (2012). Reference Method for Broth Dilution Antifungal Susceptibility Testing of Yeasts, Fourth Informational Supplement, CLSI."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Sumlu, E., Aydin, M., Korucu, E.N., Alyar, S., and Nsangou, A.M. (2024). Artemisinin May Disrupt Hyphae Formation by Suppressing Biofilm-Related Genes of Candida albicans: In Vitro and In Silico Approaches. Antibiotics, 13.","DOI":"10.3390\/antibiotics13040310"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"891","DOI":"10.1111\/j.1600-0463.2007.apm_630.x","article-title":"Quantification of biofilm in microtiterplates: Overview of testing conditions and practical recommendations for assessment of biofilm production by staphylococci","volume":"115","author":"Stepanovic","year":"2007","journal-title":"APMIS"},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Lobiuc, A., Pav\u0103l, N.E., Mangalagiu, I.I., Gheorghi\u021b\u0103, R., Teliban, G.C., Am\u0103riuc\u0103i-Mantu, D., and Stoleru, V. (2023). Future Antimicrobials: Natural and Functionalized Phenolics. Molecules, 28.","DOI":"10.3390\/molecules28031114"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"427","DOI":"10.1111\/lam.12868","article-title":"Key role of hydrogen peroxide in antimicrobial activity of spring, Honeydew maquis and chestnut grove Corsican honeys on Pseudomonas aeruginosa DNA","volume":"66","author":"Poli","year":"2018","journal-title":"Lett. Appl. Microbiol."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"2518","DOI":"10.1002\/ptr.8148","article-title":"Insights into the anti-Candida albicans properties of natural phytochemicals: An in vitro and in vivo investigation","volume":"38","author":"Yue","year":"2024","journal-title":"Phytother. Res."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"223","DOI":"10.1080\/10715760500473834","article-title":"Antioxidant and antimicrobial activities of rosemary extracts linked to their polyphenol composition","volume":"40","author":"Moreno","year":"2006","journal-title":"Free Radic. Res."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Wu, J., Wu, D., Zhao, Y., Si, Y., Mei, L., Shao, J., Wang, T., Yan, G., and Wang, C. (2020). Sodium New Houttuyfonate Inhibits Candida albicans Biofilm Formation by Inhibiting the Ras1-cAMP-Efg1 Pathway Revealed by RNA-seq. Front. Microbiol., 11.","DOI":"10.3389\/fmicb.2020.02075"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Wu, Y., Sun, A., Chen, F., Zhao, Y., Zhu, X., Zhang, T., Ni, G., and Wang, R. (2024). Synthesis, structure-activity relationship and biological evaluation of indole derivatives as anti-Candida albicans agents. Bioorg. Chem., 146.","DOI":"10.1016\/j.bioorg.2024.107293"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"707","DOI":"10.1016\/j.tim.2014.09.001","article-title":"Candida albicans hyphal initiation and elongation","volume":"22","author":"Lu","year":"2014","journal-title":"Trends Microbiol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"jkad151","DOI":"10.1093\/g3journal\/jkad151","article-title":"Strain variation in gene expression impact of hyphal cyclin Hgc1 in Candida albicans","volume":"13","author":"Sharma","year":"2023","journal-title":"G3"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1320","DOI":"10.1128\/EC.00046-10","article-title":"Candida albicans Ume6, a filament-specific transcriptional regulator, directs hyphal growth via a pathway involving Hgc1 cyclin-related protein","volume":"9","author":"Carlisle","year":"2010","journal-title":"Eukaryot. Cell"},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"McCall, A.D., Pathirana, R.U., Prabhakar, A., Cullen, P.J., and Edgerton, M. (2019). Candida albicans biofilm development is governed by cooperative attachment and adhesion maintenance proteins. NPJ Biofilms Microbiomes, 5.","DOI":"10.1038\/s41522-019-0094-5"},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Garbe, E., Thielemann, N., Hohner, S., Kumar, A., Vylkova, S., Kurzai, O., and Martin, R. (2023). Functional analysis of the Candida albicans ECE1 Promoter. Microbiol. Spectr., 11.","DOI":"10.1128\/spectrum.00253-23"}],"container-title":["Journal of Fungi"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2309-608X\/10\/11\/751\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T16:24:19Z","timestamp":1760113459000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2309-608X\/10\/11\/751"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,10,30]]},"references-count":42,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2024,11]]}},"alternative-id":["jof10110751"],"URL":"https:\/\/doi.org\/10.3390\/jof10110751","relation":{},"ISSN":["2309-608X"],"issn-type":[{"value":"2309-608X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,10,30]]}}}