Aims: To evaluate the potential effect of matrine on reducing the growth of hypha and lowering the resistance of a fluconazole-resistant colony of Candida albicans. Methods and Results: Candida albicans SC5314 and a fluconazole-resistant C. albicans 215 were used. As for C. albicans SC5314, minimal inhibitory concentration (MIC80) and effective concentration (EC50) were determined, 1 mg ml(-1) matrine could inhibit nearly 80% of planktonic growth by inverted microscope, 2 mg ml(-1) matrine suppressed 50% of metabolic activity of biofilm by XTT assay, vanishing hypha could be observed on spider agar containing 2 mg ml(-1) matrine, the expressions of three hypha-related genes, namely ALS 3, SUN 41 and PBS 2, were suppressed by 29, 45 and 61% by 2 mg ml(-1) matrine. Also, matrine could lower the resistance of C. albicans 215, in either the free-floating form or the biofilm phenotype. Conclusions: Matrine had favourable antifungal potential and might be able to reverse the fluconazole resistance of clinical isolates at relatively high concentration. The anti-candidal performance of matrine could be tightly associated with yeast-to-hypha transition proved by spider agar test and qRT-PCR. Significance and Impact of Study: More efforts are needed to find new antifungal agents. Matrine could be a potential candidate to fight against Candida-related infections by regulating yeast-to-hypha transition.