The complete mitochondrial D-loop region from 123 individuals in 12 Chinese cattle breeds and two individuals in Germany Yellow cattle breed was sequenced and analyzed. The results were shown as follows: 93 variations and 57 haplotypes were detected, and the average number of nucleotide difference was 22.708, nucleotide diversity (d) was 0.0251 +/- 0.00479, and haplotype diversity (Hd) was 0.888 +/- 0.026, indicating very high genetic diversity in Chinese cattle breeds. In the Neighbor-Joining tree, 13 cattle breeds were divided into two main clades, Bos taurus and Bos indicus; new Clade ? had only one sequence from Apeijiaza cattle breed in Tibet, which was similar to that of yak at a higher level than other cattle breeds, proving the introgression of genes from the yak. The proportions of Bos taurus and Bos indicus were 64.3% and 35.7% respectively in Xigazê Humped cattle breed, and 50.0% and 50.0% respectively in Apeijiaza cattle breed, which revealed that Tibet cattle included Bos indicus haplotypes. The importance of Yunnan cattle in the origin of Chinese cattle was also confirmed based on their abundant haplotypes. Then, a very special haplotype i1 discovered in 27 Chinese cattle breeds, including 11 breeds in this study and 16 breeds in the GenBank, played the role of a nucleus in Chinese zebu and was further discussed. At the same time, the construction of Chinese zebu core group based on haplotype i1 validated the distinct origin of Bos indicus in Tibet, which was different from that of the other cattle breeds with zebu haplotypes in China.