The marine grouper species are considered high value food fish in Southeast Asia.Aimed to get some new types of grouper,we have successfully cross-bred three species of grouper.The hybrid grouper was produced by fertilising the eggs of the orange-spotted grouper(Epinephelus coioides),tiger grouper(Epinephelus lanceolatus)with the sperm of the giant grouper(E.lanceolatus)through thein vitro fertilisation technique.The hybrids combine morphological features of the two parent species.And they are healthy and growing well in sea cages and the finding will be good news for the aquaculture industry.The genetic diversity among orangespotted grouper,tiger grouper,giant grouper and their hybrid F₁ generation population(Qinglong grouper and Hulong grouper)were tested by 6 microsatellite loci.The gene frequency,effective number of alleles(A_e),observed heterozygosity(H_o),polymorphism information contents(_PIC),genetic similarity and Nei’s genetic distances(D)were studied.The results showed that,the E.fuscoguttatusshowed the most alleles(6.849 3),while theE.lanceolatusshowed the least(2.608 6).Rich polymorphisis was observed by the six primers,and thePIC values in the five populations were 0.666 7,0.751 3,0.441 9,0.664 0,0.542 6,respectively.And the F₁ generations showed the highest observed heterozygosity(H_o)of 0.940 0,while theE.lanceolatus showed the lowest value of 0.508 3.The results of genetic distances and genetic similarity showed that both F₁ generations had a close relationship with the male parent E.lanceolatus.The alleles of F₁ generations were mostly generated from both of their parents,which mean that the genetic materials were inherited from parents.And the higher genetic diversity that existed in the two hybrids indicated potential heterosis.