Significant correlations have been found between the nitrate concentration in young corn (Zea maysL.) stalks and available soi 1 N and relative grain yield when N availability is a yield limiting factor. To successfully use nitrate tissue testing as an indicator of the N status of plants, the effects of factors other than soil or fertilizer N on the nitrate concentration in the tested tissue must be understood. Diurnal effects on stalk nitrate content were studied in a greenhouse experiment, in which stalk samples were collected every two hours for 24 hours, and in a field study, where plants in an N‐fertilizer response experiment were sampled at sunrise and at mid‐day. A significant diurnal cycle was observed for stalk nitrate content in the greenhouse, with a peak at 0800 h and a low at 1400 h, but no diurnal effect was found in the field study, possibly due to plant stress from other environmental factors. In a second greenhouse study, stalk samples from seedlings in direct and 66% shaded natural light were collected for two consecutive days and measured for nitrate content to ascertain the consequences of sampling on overcast days. Significant accumulations of nitrate occurred in stalks of plants sampled after two consecutive days of shading. Stalk samples from three standard hybrid corn trials in central Pennsylvania were measured for nitrate content to determine if hybrid differences existed. Hybrid differences in stalk nitrate were significant in only one of the three sites studied, and the variation in that one may have been due to differences in drought tolerance as well as nitrate metabolism. The results suggest that sampling strategies should be designed to minimize the effects of irradiance level on nitrate accumulation, but that differences in nitrate accumulation among corn varieties may not be a major concern in using nitrate tissue tests to predict N availability.