In this paper, the HKG method studied and analysised the C-banding of chromosomes in triple cross of ‘Inner Mongolia Hibrid oil sunflower 3’were used.The results showed that every chromosome had at least one C-band, and there were 62 C-bands on the chromosome pairs, most of them were centromeric and interbands, and the interbands were mainly distributed on the short arms; C-band intensity were different , 46 were strong and 16 were weak. The C-bands formula was 2n=2x=34=8I++3T++5I+I+T++4C+2CI+4CI++3CI++I+T++CT++2CT+ Each chromosome had its own unique band pattern distinguishable from all the others, so each chromosome of sunflower can be distinguished by Giemsa C-banding.