| Primers of CHD gene and primer combinations of EE0.6 sequence were used to identify the sex of Grus vipio, Anthropoides virgo, Balearica pavonina, Grus japonensis, Grus grus with the technique of PCR and electrophoresis. In order to find out the condition of genetic diversity and phylogenic relationship among 42 individuals of 4 crane populations (Grus vipio, Anthropoides virgo, Grus japonensis, Grus grus), the universal primer of control region of mtDNA was used to amplify the extracted DNA, and several sequences were obtained. Two specific primers were designed from the sequences and their amplifications were sequenced too. Genetic diversity of these cranes and phylogenic relationship among four crane populations were analyzed. The main results were summarized as following:1. The primer 2550F/2718R of CHD gene and nine primer combinations of EE0.6 sequence were used to amplify the gene or sequence, the results showed that there were two bands in female and one band in male, which can successfully identify the sex of Grus vipio, Anthropoides virgo, Balearica pavonina, Grus japonensis, Grus grus and avoid the disturbance of the false negative.2. 605 bp sequence of mtDNA control region of 42 individuals of 4 crane populations were sequenced and analyzed. The result showed the content of nucleotide A, C, G, T were 28.80%,23.80%,14.50% and 32.90%, respectively. The percentage of A+T was 61.70% which was higher than G+C content. There were 79 polymorphic sites represent 13.72% of total analyzed sites, in which 20 sites were singleton polymorphic sites and 59 sites were parsimony informative sites. Seven kinds of variance were found in this region, such as transition, transversion, indel, transition and transversion in the same site, indel and transition in the same site, indel and transversion in the same site, indel and transition, transversion in the same site. The ratio of transition and transversion in this study was 0.09. There were 33 haplotypes, each population had its own haplotypes and no sharing haplotypes. The distribution of all haplotypes among the populations was disequilibrium and the diversity of haplotypes was ranged from 0.889 to 1.000.The total diversity of haplotypes was 0.974±0.018, indicated that there existed rather abundant mitochondrial genetic diversity in 4 crane populations.The Grus vipio showed the highest diversity of nucleotide, 0.684%. The average number of nucleotide divergence (K) and average nucleotide diversity (Pi) were 23.941 and 4.156% respectively, which indicated the high level of diversity among populations. Interspecies Nucleotide Divergence (Dxy) in 4 crane populations was ranged from 5.225% to 6.649%, whereas Interspecies Net Nucleotide Divergence (Da) was ranged from 4.847% to 6.270%. Kimura 2-parameter distance among these populations ranged from 0.054 to 0.070. Analysis of molecular variance showed that 93.86% of genetic variation was present between populations. FST value was 0.93863, which indicated the genetic variation was significant between populations (P < 0.01). There were significant divergence among the 4 crane populations.3. The NJ, ME and UPGMA phylogenic dendograms of 33 haplotypes in 4 crane populations were constructed. As a whole, the results of three kinds of trees are same and these 33 haplotypes were placed into four lineages. Each lineage contained the specific haplotypes of each crane population. The median-joining networks of the 4 crane populations in the control region also showed the same results with phylogenic trees and all 33 haplotypes appeared into four clusters.4. Neutrality test with the value of Tajima's D of control region of mtDNA showed that except Anthropoides virgo which showed significant difference from neutrality, the other three populations were accord with the neutrality.
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