Pecan (Carya illinoinensis) is an important woody oil crop. 'Pawnee', 'Mahan', 'Jinhua', 'Zhongshan25' are all fine varieties that have been promoted to cultivate in large scale in China. The chromosome numbers and karyotypes of four pecan cultivars were studied by applying traditional squash technique. The results showed that the chromosome number of all tested cultivars was 32. The karyotypes of four kinds of Carya illinoinensis were 2B. The results indicated that the karyotype formula of 'Pawnee' was 2n=32=12sm+18m+2T, chromosome composition of relative length was 2S+16M1+12M2+2L; and the karyotype formula of 'Mahan' was 2n=32=16sm+14m+2T, chromosome composition of relative length was 6S+12M1+8M2+6L; as to 'Jinhua', the karyotype formula was 2n=32=10sm+20m+2T, the chromosome composition of relative length was 4S+12M1+14M2+2L, the last one is 'Zhongshan25', the karyotype formula was 2n=32=8sm+22m+2T, the chromosome composition of relative length was 6S+12M1 +8M2+6L. These index of kary otype asymmetries of four pecan cultivars were 62.07%, 64.68%, 62.47%, 62.38%. 'Pawnee' was relative original while 'Mahan' was evolutional during genome evolution. This study could provide cytological basis for the identification and breeding of germplasm resource of pecan.