Outbreaks of leaf spot disease occurred in Rehmannia glutinosa fields in Henan Province, China, in 2019, with the incidence ranging from 20% to 40%. R. glutinosa plants with diseased leaves were collected, and 25 isolates were obtained. Pathogenicity tests, morphological observations, and phylogenetic analyses were conducted to identify the pathogens, and the biological characteristics and control agents of the pathogens were studied. Five isolates of pathogenic fungi were isolated. Three isolates were identified as Fusarium equiseti, which is a new pathogen causing R. glutinosa leaf disease; the other two isolates were identified as Fusarium acuminatum. The mycelia of F. equiseti grew fastest on Czapek medium, and the optimal temperature and pH were 25 °C and 10.0, respectively. The mycelia of F. equiseti grew from 5 °C t o 35 °C, and the lethal temperature was 55 °C. The optimal carbon and nitrogen sources were soluble starch and peptone, respectively. Eight fungicides had inhibitory effects on the mycelial growth of F. equiseti and F. acuminatum. Prochloraz had higher activities against F. equiseti and F. acuminatum, with EC₅₀ values of 0.139 mg·L⁻¹ and 0.123 mg·L⁻¹, respectively. These results provide useful information that will aid the development of management strategies to control leaf diseases of R. glutinosa caused by F. equiseti and F. acuminatum.