Isolation Of Microsatellite Makers For Leuciscus Leuciscus Baicalensis From The Irtysh River In Xingjiang And Analysis Of Its Genetic Diversity

In the present study,we developed 30 novel microsatellite makers focused on Leuciscus leuciscus baicalensis sampled from the Irtysh River in Xingjiang using next-generation sequencing technology and capillary electrophoresis,from which 19 was genomic microsatellite makers and 11 was expressed sequence tag(EST)microsatellite makers.Primers were used to test genetic diversity in three L.leuciscus baicalensis populations(the Habahe River population,the Buerjin River population and the Beiwan section population).In the present study we also tested the cross utility of polymorphic microsatellite makers mined from L.leuciscus baicalensis in nine other Cyprinidae fish species to investigate transferability.The results of this research were summarized as follows:1.In total,160 pairs of primers(64 from genomic libraries and 96 from EST libraries)were designed and amplified using the DNA from the BW samples,in which three pairs isolated from the genomic libraries and 25 pairs isolated from the EST libraries failed to provide PCR products or the expected products.2.Genetic diversity was assessed using 30 polymorphic markers in three populations,339 alleles(Na)were found,varied from 4 to 27(with an average of 11.3).The number of effective alleles(Ne)was 154.28,varied from 1.56 to 15.34(with an average of 5.14).The expected heterozygosity(He)was 0.36-0.94(with an average 0.75 � 0.14),and the observed heterozygosity(Ho)was 0.37-1.00(with an average 0.68 � 0.18).The polymorphism information content(PIC)was 0.31-0.93(with an average 0.71 � 0.15).3.There were almost non-differentiated among the three populations according to the genetic distances and F-statistic.Almost all the variation was between individuals,very little variation occurred among populations.4.The rate of polymorphism was higher in genomic microsatellite markers(31.1%)than that in the EST microsatellite markers(15.5%).The average values of PIC,He,Ho,Ne and Na for the EST microsatellite markers were lower than those for the genomic microsatellite markers.5.The highest transferability was in Rutilus lacustris(80%),followed by Leuciscus idus(76.7%).The EST microsatellite markers were more transferable than the genomic microsatellite markers.