Culturable airborne microorganisms significantly impact air quality and human health in urban forest land. Their concentrations serve as key air quality indicators. Over a year, this study analyzed airborne microorganisms in six forest stands within the Zhuyu Bay Scenic Area, Yangzhou, Jiangsu Province, China, to assess concentration characteristics and seasonal variations. Results showed that bacterial concentrations peaked in spring and summer, while fungal concentrations were highest in March. Microbial levels remained elevated from April to June, with variations among forest stands. A correlation analysis linked humidity, temperature, negative ion concentration, particulate matter 2.5 (PM2.5), and air pressure to microorganism fluctuations. To further explore the impact mechanism of urban microclimate on air microorganism concentrations, this study confirmed a strong positive influence of climatic factors on microorganism concentrations, particularly temperature and humidity. In conclusion, this study identifies seasonal patterns and microclimate interactions that affect airborne microorganism concentrations in urban forests. Findings contribute to ecosystem assessment, urban ecological planning, and climate improvement strategies, supporting informed decision-making.