Genetic Diversity Of Hipposideros Pratti In Henan Province Based On RAPD And Microsatellite Markers

Hipposideros pratti belongs to Hipposideridae,Chiroptera,which distributed mainly in central and southern provinces of China,H.pratti of Henan Province is only distributed in Nanyang.H.pratti belongs to typical cave-dwelling bat,many caves where H.pratti inhabits have been developed as places of tourist attraction,which seriously affect the survival of this species.On account of the killing of bats,abuse of pesticide and insecticide and other factors,the number of H.pratti decreased dramatically.For the purpose of mastering the genetic diversity of H.pratti,and providing scientific management and protection for them,using RAPD and microsatellite primers to research the genetic diversity based on 74 samples from four counties(Xixia,Neixiang,Nanzhao,Junma)of Henan Province,analyzed the genetic differentiation and genetic structure of H.pratti,then tested the population whether experience significant bottleneck effect.The results are shown as follow:1.Eight RAPD primers and six microsatellite primers which selected from the published sequences were used to amplify four populations of H.pratti in Henan province,91 DNA bands were amplified by eight RAPD primers in all,the average polymorphic locus(P)was 50.00.25 alleles were detected by six microsatellite primers in all,with the value of mean allele of 4.17 detected for each locus,the value of average PIC of the six loci was 0.498.2.Based on the data of RAPD,four population of H.pratti on the number of observed alleles(N_a) of H.pratti ranged from 1.43 to 1.62,the number of effective alleles(N_e)ranged from 1.33 to 1.49,the number of Shannon information index(I)ranged from 0.2625 to 0.3805.Based on the data of microsatellite,the number of observed alleles(N_a)of H.pratti ranged from 2.83 to 3.83,the number of effective alleles(N_e)ranged from 2.03 to 2.79,the frequency of invalid alleles ranged from 0.017 to 0.159,the number of Shannon information index(I)ranged from 0.7955 to 1.0996.The data RAPD and microsatellite showed consistent results,four populations have different genetic diversity,Nanzhao(NZ)had the highest genetic diversity,while Neixiang(NX)had the lowest genetic diversity.3.Based on the data of microsatellite,the value of observed heterozygosity(H_o)of four populations of H.pratti was from 0.298 to 0.531,the value of expected heterozygosity(H_e)was from 0.422 to 0.610,the value of Nei's expected heterozygosity ranged from 0.412 to 0.595,the value of polymorphic information content(PIC)was from 0.382 to 0.541 The results showed that inbreeding phenomenon existed in every population,the polymorphism of Nanzhao(NZ)was highest(PIC?0.5),while the polymorphism of Xixia(XX),Junma(JM),Neixiang(NX)were moderately(0.25?PIC?0.5).The Hardy-Weinberg test revealed that 16 times of the results deviated from the Hardy-Weinberg equilibrium based on microsatellite.4.The F-statistics of the population of H.pratti were calculated based on microsatellite data,the results showed that F_is ranged from-0.0754 to 0.3714,F_it ranged from 0.0064 to 0.4245,it could speculate that inbreeding phenomenon was found in all four populations(F_is?0,F_it?0).5.Population differentiation level was analyzed based on RAPD and microsatellite data,the population genetic differentiation coefficients(F_st)of H.pratti were 0.2236 and 0.0614 respectively.Respectively,the proportion of genetic variation among four populations was 77.64%and 75.39%of total genetic variation.The value of mean gene flow for four H.pratti populations was 1.7364 and 3.8206respectively.That means the genetic variation among four populations was the main factor causing the population variation of H.pratti,and according to the research results,there was a certain level of gene flow among four populations(Nm?1).The degree of genetic differentiation of H.pratti based on RAPD was highly genetically differentiated,while the results based on microsatellite markers was moderately,indicating that a variety of molecular markers are evaluated the genetic status more comprehensively and accurately in the study of population genetic diversity.6.Analyzed genetic structure of populations based on RAPD and microsatellite data,the results showed the Nei's genetic consistency(GI)between two populations of H.pratti ranged from 0.9002 to 0.9456,0.8834 to 0.9578 respectively.The genetic distance(GD)ranged from 0.0559 to 0.1051,0.0431 to 0.1240respectively.Neixiang(NX)and Junma(JM)had the closest genetic relationship,while Xixia(XX)and Nanzhao(NZ)had the farthest ties of consanguinity,and the overall genetic differentiation among four groups was small.7.Based on microsatellite data,analyzed whether the population of H.pratti experience a bottleneck effect,the results showed that under the model of IAM,TPM and SMM,only Xixia(XX)experienced significant bottleneck effect(P?0.05).While under the first two models,Nanzhao(NZ)and Xixia(XX)experienced significant bottleneck effect(P?0.05).Under the two models of TPM and SMM,the heterozygosity of Neixiang(NX)was deficiency(He?Hd),under the three models,the heterozygosity of Xixia(XX),Nanzhao(NZ),and Junma(JM)were excess(He?Hd),the model of gene distribution frequency was L-shaped.To sum up,the eight RAPD primers and six microsatellite loci selected had polymorphism.There had inbreeding behavior within population.Among populations,there occurred genetic differentiation and the genetic distances from different group were all different.Among the four populations,Nanzhao(NZ)had the highest genetic diversity,followed by Junma(JM)and Xixia(XX),and Neixiang(NX)had the lowest genetic diversity,the main factor of group variation was genetic variation among populations.The significant bottleneck effect was detected in Nanzhao(NZ)and Xixia(XX),it is necessary to manage populations of H.pratti scientifically and effectively.