Aiming at spectral detection of apple fluorescence canopy, the present paper carried out spectral detection tests under different weather conditions, different detection times, and different detection heights and angles to apple canopy in the two years of 2008 and 2009, so as to analyze impacts of these factors on apple canopy spectral characteristics and explore standardized spectral detection methods for apple fluorescence canopy. The results indicated the regularity in spectral reflectance of apple fluorescence canopy to a certain degree under different conditions, especially in the 760-1 350 nm near-infrared bands. The authors found that canopy spectral reflectance declined along with the decrease in sunshine and it is appropriate to detect canopy spectrum in sunny days with few clouds. In addition, spectral reflectance tended to be stable when the wind scale was below grade 2. The discrepancy of canopy spectra is small during the time period from 10:00 to 15:00 of a day compared to that of other times. For maintaining stable spectral curves, the height of detector to apple canopy needed to be adjusted to cover the whole canopy within the field of view according to detection angle of the detector. The vertical or approximately vertical detection was the best for canopy spectral reflectance acquisition. The standardization of technical methods of spectral detection for apple fluorescence canopy was proposed accordingly, which provided theoretical references for spectral detection and information extraction of apple tree canopy.