Abstract: The pathogenic fungi isolated from the decayed and non-symptom seeds of Chinese chestnut in Luotian, Hubei, China, identified into 11 species by morphological characteristics and phylogenetic characteristics based on ITS rDNA sequences. The results showed that those pathogenic fungi were Rhizopus stolonifer、Fusarium solani、Stachybotrys chartarum、Cryphonectria parasitica、Lasiodiplodia theobromae、Penicillium expansum、Aspergillus niger、Colletotrichum gloeosporioides、Cryptococcus laurentii、Cryptococcus luteolus、Candida albicans; the isolating rates of Cryphonectria parasitica and Penicillium expansum were higer than other isolated pathogenic fungi. Tablet confrontation test of 6 antagonistic bacteria strains showed that pathogenic fungi of Pseudomonas syringae and Pseudomonas fluorescens had remarkable antimicrobial activities to pathogenic fungi isolated from Chinese chestnut, Pseudomonas fluorescens showed higher resistant to pathogenic fungi TIS4 and ITS6 than Pseudomonas syringae.