Cryptomeria japonica D. Don is a coniferous tree species widely grown in southern China for its high ornamental value. Recently, during disease surveys in China, a symptom of dieback occurred on C. japonica in Nanjing, Jiangsu Province, China. A total of 130 trees were surveyed and more than 90% showed the same symptom. The crowns of affected trees were brown when viewing from a distance, and the bark showed no difference from the healthy ones. In this study, 157 isolates were isolated from the 3 affected plants of C. japonica, and based on the living culture on PDA, the fungal isolates were preliminarily divided into 6 groups. Thirteen representative isolates were selected for the pathogenicity test, and seven of them showed obvious pathogenicity on C. japonica, causing stem basal canker. These isolates were identified based on comparisons of the DNA sequences of the internal transcribed spacer regions (ITS), partial translation elongation factor 1-alpha (tef1), β-tubulin (tub2), and DNA-directed RNA polymerase II subunit (rpb2) and combined with their morphological characteristics. Results showed that these seven isolates belong to two taxa in Neofusicoccum, including a species new to science. The new species, Neofusicoccum cryptomeriae, was hereby described and illustrated. The other species was N. parvum. Both species were pathogens of stem basal canker of Cryptomeria japonica.