Thirteen microsatellite markers of Epinephelus awoara previously discovered by our lab were selected to analyze the genetic diversity and phylogenetic relationship of nine species of grouper (E. awoara, E. merra, E. fario, E. fasciatus, E. lanceolatus, E. akaara, E. septemfasciatus, E. coioides and E. fuscoguttatus) from South China Sea. The results showed that the number of total alleles of these 13 microsatellite loci was 84 in these fishes, the mean number of alleles ranged from 2.69 to 5.38, mean polymorphism information content (PIC) ranged from 0.1976 to 0.4267, mean observed heterozygosity (Ho) from 0.4615 to 0.6239, mean expected heterozygosity (He) from 0.3510 to 0.4754 and mean Hardy-Weinberg departure value (D) from 0.1097 to 0.2836, respectively. All of these indicated that genetic diversity of the nine species of grouper was at a medium level. Two NJ dendrograms showed that E. coioides, E. fuscoguttatus and E. lanceolatus were grouped together, while E. awoara, E. akaara and E. septemfasciatus were in a second group, and E. merra, E. fasciatus and E. fario were in a third group which had a relatively closed relationship with the second group. The dendrograms could also support a conclusion that Promicrops lanceolatus (E. lanceolatus) should be included in genus Epinephelus.