Isolation Of Microsatellite Markers And Analysis Of Population Genetics In Hipposideros Armiger

Order Chiroptcra is one of the most widely distributed and largest order of mammals,and is also one of the most enigmatic groups of mammals.They are the only mammals that have achieved true self-powered flight.Hipposideros armiger belongs to Microchiroptera,and has a wide Asian geographical distribution that includes S and SE of China.They play an important role in ecosystem,and have a close relationship with human.In recent years,because of habitat destruction and human direct disturbance resulted from increased human activities,the population structure of Hipposideros armiger is seriously affected.Studies on this species have been carried out on the behavior ecology,predation and echolocation calls and karyology analysis,very few research of molecular ecology has been reported.Microsatellite DNA has been widely used in study population genetics as an effective molecular tool.In order to study the population genetics of Hipposideros armiger,we characterized eight microsatellite markers from genome of Hipposideros armiger using the microsatellite enriched method.We selected five polymorphic microsatellite loci,and four loci from Hipposideros pratti according to cross-amplification in related species.13 populations and 250 individuals were genotyped with these nine microsatellite loci.Genetic diversity analysis revealed:1) A total of 163 alleles were identified,number of mean alleles was 4.89-11.67. Allelic richness and observed heterozygosity of continent were higher than Hainan and Taiwan.Allelic richness was the highest in Guizhou(5.18) and lowest in Xinzhu (3.55),and the observed heterozygosity was the highest in Guizhou(0.88),the lowest in Hainan(0.61).Tests of Hardy-Weinberg equilibrium showed just 3%were significant by permutations following Bonferroni correction.No linkage disequilibrium was detected between locus pairs across populations.2) Allelic richness values were positively correlated with latitude of sampling sites and negatively correlated with longitude of sampling sites when considering all populations.The data showed that the population of Hipposideros armiger colonized from west to east and from mainland to island.3) The analysis of population differentiation showed significant genetic differentiation across most of populations(P＜0.001).Significant genetic differentiation was detected across the population of mainland,Hainan and Taiwan. The gene flow between populations of Hipposideros armiger was very limited. Population differentiation followed a pattern of isolation by distance across all populations(b = 0.041,a = -1.83,R~2=0.23,P＜0.001).A pattern of isolation by distance across all populations of mainland was not detected.The result showed that geographic barriers played an important role in population genetic differentiation. The correlation between allelic richness and longitude suggested that protecting habitat in lower longitude areas is more significant and effective to Hipposideros armiger.