The smut Ustilago esculenta is an endophytic fungus that invades Zizania latifolia (Jiaobai) and plays an important role in inducing the host stem to form a swollen gall. The aims of this study were to investigate isolation, identification and inoculation methods of U. esculenta from Jiaobai. The spore suspension culturing method was the best method of isolating U. esculenta, and yeast-like fungal strains and septate hyphal strains were obtained. The universal primers ITS1 and ITS4 amplified a ribosomal DNA spacer element from all tested strains, while specific primers for Lam16A gene generated a 442 bp product only with septate hyphae strains. De novo transcriptome assembly results showed that the yeast-like fungal strains did not match Ustilago, and genomic analysis suggested the septate hyphae strains were highly similar to known smut fungi. Therefore, the septate hyphae strains were identified as U. esculenta, which indicated that the ITS primers were not species-specific but that Lam16A-specific primers can distinguish U. esculenta accurately and easily. In vitro, U. esculenta grew quickly on PDA medium at 28 °C in darkness. Furthermore, an inoculation method was established to produce swollen stems by injecting a spore suspension of U. esculenta at OD₆₀₀ = 2.5 into the basal culm internode of male Jiaobai.

黑穗病Ustilago esculenta是一种内生真菌，侵入茭白（Zizania latifolia），在诱导寄主茎形成肿瘿中起重要作用。本研究的目的是研究来自胶白的美国香茅的分离、鉴定和接种方法。孢子悬浮培养法是分离紫菜的最佳方法，获得了酵母样真菌菌株和有隔菌丝菌株。通用引物 ITS1 和 ITS4 从所有测试菌株中扩增出一个核糖体 DNA 间隔元件，而Lam16A基因的特异性引物仅产生 442 bp 的产物。从头开始转录组组装结果显示酵母样真菌菌株与黑粉菌不匹配，基因组分析表明有隔膜菌丝菌株与已知的黑穗病真菌高度相似。因此，有隔的菌丝菌株被鉴定为U.星虫，这表明ITS引物为未物种特异性但Lam16A特异性引物可以区分U.星虫准确且容易地。在体外，U. esculent a 在 PDA 培养基上在 28 °C 的黑暗中快速生长。此外，建立了一种接种方法，通过将OD ₆₀₀  = 2.5的U. esculenta孢子悬浮液注入雄性交白的基部秆节间来产生肿胀的茎。