Development Of Microsatellite Loci In Transcriptome Of Forest Musk Deer And Phylogenetic Analysis

Forest musk deer(Moschus berezovskii)belongs to Artiodactyla,Ruminantia,Moschus,Moschus berezovskii.It is the smallest animal in Moschus and famous for its musk which excreted by adult males.Due to the huge economic value of musk,forest musk deer has been poached on a large scale,the population of forest musk deer has decreased sharply.In 2002,forest musk deer was listed as a national first-class key protected animal.Besides,forest musk deer was listed as endangered on the 2008 IUCN Red List of threatened species.At present,artificial cultivation of forest musk deer is the main means to protect its germplasm resources and musk acquisition.However,due to the single pursuit of the number of forest musk deer and blind breeding,results in the decrease of genetic diversity and individual somatoplasm of musk.In order to enrich the genetic data of musk deer,provide guidance for the breeding of musk deer,and enrich the polymorphic sites of musk deer.The following studies were carried out in this paper: the bone marrow cells from dead musk deer were extracted for culture,and the c DNA library was obtained from it,then the transcriptome sequencing was carried out to obtain the microsatellite sites of musk deer;Nine polymorphism microsatellite loci were obtained from 100 microsatellite loci with verification.Finally,we used these 9 microsatellite loci to analyze the genetic diversity and cluster analysis of 15 musk deer samples from Feng Chun musk deer farm.The main results are as follows:1.Bone marrow cells were successfully isolated from the femoral head of dead forest musk deer and cultured to P6 generation.2.The RNA of musk deer was extracted from the bone marrow cells,and the c DNA library of musk deer was obtained after inversion.According to the analysis of transcriptome sequencing,100 pairs of primers for musk deer microsatellite loci were selected.Using agarose gel electrophoresis for specific detection,49 pairs of primers in 100 pairs of primers were specific.3.49 pairs of primers were tested for polymorphism,and 9 polymorphic loci were obtained.4.Polymorphisms and genetic relationships of 15 introduced male forest musk deer were detected by using these 9 microsatellite loci,and found that the population of 15 forest musk deer could be grouped into 8 groups with distant genetic relationships,among which the forest musk deer numbered 11539 and 08562 were the most distant from other forest musk deer branches in the forest musk deer population.The results of this study proved that transcriptase sequencing is an effective method to obtain microsatellite loci of forest musk deer.The microsatellite markers obtained in this study can also be used to reveal the genetic relationship of forest musk deer,which is conducive to screening high-quality individuals in the breeding of forest musk deer,increase the genetic diversity of the population,and provide reference for the subsequent identification of musk deer kinship.