The spectrum and physical-chemical parameters were measured on cotton leaves infected by aphid with different severity levels (SL) at main cotton growth periods. Meanwhile, the reflectance and physical-chemical parameters of cotton leaves infected by aphid were analyzed and compared in different cotton growth periods and varieties. The sensitivity wave bands of cotton leaves infected by aphid were confirmed, and the estimating models of leaves infected by aphid were established. The results showed that there was a significant difference in the spectrum and physical-chemical parameters of cotton leaves infected by aphid. The thickness, water and Chl.b increased, while Chla, Chl.a+b and Cars content decreased in leaves infected by aphid. Besides, in visible region, the reflectance of cotton leaves infected by aphid has shown going up first and then down in different cotton growth periods and varieties with increasing of SL. However, in NIR region, it has shown discrepancy in varieties. The 434-727 and 648 nm can be used as sensitive and optimal aphid-band for cotton leaves. Estimation models for leaves infected by aphid were all in significant correlation. Among all the models, the model of (R1 589-R648)/ (R1 589+R648) had the best estimation precision, RE was the smallest (0.128), and it was commended as best models to estimate SL of leaves infected by aphid. The study provides an experimental reference for monitoring spectrum of cotton infected by aphid with remote sensing in large areas.