Negative oxygen concentration has become an important indicator to measure air quality. Choosing 17 national meteorological stations in Hubei on behalf of the observation stations with the same environment and using the equipment in line with national and industry standards, the spatialtemporal characteristics of the air negative oxygen ion from March to May in 2014 and its relationship with environmental and meteorological factors are analyzed following the definition of negative oxygen ion. The results show that negative oxygen concentration in the western Hubei is higher than that in middleeast, and the average concentration of the Shennongjia forest district is the highest, but it is low in Wuhan, the central of the middleeast. Over the span of a whole day, it is high early in the morning, but low during the day, and then it has a trend of rising in the evening. It is also reveals that the concentration correlates positively with altitude and vegetation coverage, and negatively with small particles in the air. However, the concentration has poor correlates with temperature, air pressure, relative humidity, speed, sunshine duration, and other meteorological factors. Besides, the precipitation and thunderstorm is favorable to the increase of negative oxygen ions.