Genetic Diversity Of Pipistrellus Abramus In Henan Province Based On Microsatellite Markers

Pipistrellus abramus belongs to Pipistrellus of Vespertilionidae in Microchiroptera,it is distributed in eastern Asia abroad,in China,it is distributed in Heilongjiang,Liaoning,Shanxi,Hebei,Sichuan,Yunnan,Shandong,Gansu,Anhui,Zhejiang,Hubei,Hunan,Guangxi,Fujian,Taiwan and Hainan,and the whole province of Henan.P.abramus is a typical house-type bat.In recent years,the species has seen its numbers plummet due to habitat loss as concrete-structured bungalows replace old-fashioned brick houses,hunting by humans and the overuse of pesticides and pesticides.So far,there is no research on the genetic diversity of Chiroptera animal in Henan Province,and the current status of genetic diversity of P.abramus is unclear.In this study,direct sequencing method was used to identify P.abramus species,and microsatellite molecular markers was used to research the genetic diversity of 105 P.abramus populations(SQ,ZK,ZMD and XX)in Henan Province.Analyzed the levels of genetic diversity,degree of genetic differentiation,the source of genetic variation and genetic structure characteristics of four P.abramus populations,and tested whether their populations experienced bottleneck effects,with a view to protecting the species and further research provides basic information.The results of the study are as follows:1.Used the direct sequencing method of mitochondrial ND1 gene fragments,a total of 105 P. abramus individuals were identified from bat specimens collected from SQ,ZK,ZMD and XX.2.Six microsatellite loci of P.abramus were selected to analyze the genetic diversity of four P. abramus populations.A total of 90 alleles(N_A)were obtained,with an average of 15 alleles(N_A)detected at each locus,and the number of alleles(N_A)at each locus was 5 to 23;the average number of effective alleles(N_e)was 5.551 and the range was between 2.383 and 8.990.The average observed heterozygosity(H_O)was0.610,ranged from 0.386 to 0.818;the average expected heterozygosity(H_e)was 0.791,ranged from 0.584to 0.894.The average polymorphism information content(PIC)was 0.761,ranged from 0.510 to 0.880,all greater than 0.5.They were highly polymorphic loci,indicated that the 6 microsatellite loci choose were rich in genetic diversity information.3.The average number of alleles(N_A)of four geographic populations of P.abramus was 5.833?11,and the average number of effective alleles(N_e)was 4.3555?5.591.The observed heterozygosity(H_O)was 0.583?0.704,and the average was 0.622;the expected heterozygosity(H_e)was 0.738?0.801.and the average was 0.622.The observed heterozygosity of each population was lower than the expected heterozygosity,indicated that there was a certain inbreeding phenomenon in the population.The average polymorphism information content(PIC)was 0.718,ranged from 0.691 to 0.753,all greater than 0.5,showed a high degree of polymorphism,indicated that the four P.abramus populations have a high degree of genetic diversity.Of the 24 Hardy-Weinberg balance tests,11 were extremely significant diverged from balance(P<0.01),and 2 were significantly diverged from balance(P<0.05).4.F-statistics and gene flow analysis of P.abramus showed that the inbreeding number(F_IS)of each site ranged from-0.0366 to 0.4413,with an average of 0.1716.The inbreeding coefficient(F_IT)of the total population was 0.0330?0.4768,with an average of 0.2204,indicated that there was inbreeding occurred in the population(F_IS>0,F_IT>0).The average gene flow(Nm)was 3.9932,and the range was 2.9267?6.1613.The gene flow(Nm)was all greater than 1,and there were frequent gene flows among different groups,which can effectively resist the genetic differentiation between groups.5.The analysis of the genetic differentiation index(F_ST)among P.abramus populations showed that the genetic differentiation index(F_ST)among different populations was between 0.01680 and 0.07861,among which there was a medium level between SQ population and XX population,and between SQ and ZMD population.Genetic differentiation(0.05ST<0.15),F_ST values were 0.07861 and 0.05099,respectively;the level of differentiation among other populations was low(F_ST<0.05).AMOVA analysis of variance showed that genetic variation between individuals(76.188%)was the main cause of variation in the entire P.abramus population.6.Construction of UPGMA cluster analysis chart based on Nei's genetic distance of P.abramus showed that the SQ population and ZK population clustered first into a branch,then into a branch with ZMD species,these three populations gathered together with the XX population.The genetic distance between populations conforms to the geographic distribution characteristics of populations.7.Bottleneck effect test results showed that XX population experienced significant Bottleneck effect(P<0.05).In conclusion,the six microsatellite loci screened were all highly polymorphic loci,and the four P. abramus populations all had high degree of genetic diversity.There was inbreeding within populations,and there were different degrees of genetic differentiation among populations.The main variation came from inter-individual variation.The genetic distance between populations conformed to the characteristics of population geographic distribution.The XX population has experienced a significant bottleneck effect.