Chlorophyll content is an important index to evaluate the photosynthetic efficiency in rice. Realtime and nondestructive monitoring of chlorophyll content are very significant to rice growth diagnosis. The quantitative relationship between hyperspectral index and chlorophyll content was analyzed using the rice P88S (green leaf) and Huang 1S (yellow leaf) as the test material. Canopy reflectance spectra and quantitative models of chlorophyll content in rice leaves were constructed. The results showed that the canopy spectral reflectance of different leaf color of rice changed with the growth of the plant. There was a highly significant positive correlation between the chlorophyll content and the first order differential spectrum in the visible wavelength range of 502711 nm for the P88S and 487716 nm for the Huang 1S. P88S RVI (363，675) and Huang 1S DVI ′(639，680) were used as spectral parameters to construct the estimation model with chlorophyll content, which was the best fitting effect. The method of hyperspectral spectroscopy combined with first order differential spectroscopy was feasible to estimate the chlorophyll content in rice.