Abstract Background The characteristics of light source have an important influence on the measurement performance of canopy reflectance spectrometer. The size of the effective irradiation area and the uniformity of the light intensity distribution in the irradiation area determine the ability of the spectrometer to express the group characteristics of the measured objects. Methods In this paper, an evaluation method was proposed to theoretically analyze the influence of the light intensity distribution characteristics of the light source irradiation area on the measurement results. The light intensity distribution feature vector and the reflectance feature vector of the measured object were constructed to design reflectance difference coefficient, which could effectively evaluate the measurement performance of the canopy reflectance spectrometer. By using self-design light intensity distribution test system and GreenSeeker RT100, the evaluation method was applied to evaluate the measurement results. Results The evaluation results showed that the vegetation indices based on the arithmetic average reflectance of the measured object could be obtained theoretically only when the light intensity distribution of the light source detected by the spectrometer was uniform, which could fully express the group characteristics of the object. When the light intensity distribution of the active light source was not uniform, the measure value was difficult to fully express the group characteristics of the object. And the measured object reflectance was merely the weighted average value based on the light intensity distribution characteristics. Conclusions According to the research results of this paper, sunlight is the most ideal detection light source. If the passive light source spectrometer can improve the measurement method to adapt to the change of sunlight intensity, its measurement performance will be better than any active-light spectrometer.