| The purpose of this study was to develop microsatellite markers for the genome of Scleropages formosus,and to evaluate the amplification efficiency effectiveness of these primers for other fish in the family Scleropages.Based on the results of genome-wide sequencing,the number and distribution of microsatellites in the whole genome of the endangered species of Scleropages formosus were analyzed by bioinformatics method.Fifty pairs of microsatellite markers were developed.The parental populations and four parental populations of cultured Scleropages formosus were analyzed by using these markers.The progeny population was identified by genetic relationship,and the progeny parents were identified to establish pedigree.1.At the same time,The genetic diversity of the cultured population was analyzed.The results showed that there were 354,239 micrositellite sequences with1-6 microsatellite repetitions,and the overall length was 6,139,487 bp.The preponderance of mononucleotide,dinucleotide,trinucleotide,tetranucleotide and pentanucleotide were A,AC,AAT,ATCC and AAAAT,respectively.Among the 25 chromosomes,chromosome 6 contained the largest number of microsatellites(28662),and the chromosome 22 had the least(6929).Linear correlation(r=0.999)was observed between the number of microsatellites and the length of DNA sequences they located in.However,chromosome 22 had the highest frequency of microsatellite occurrence(576.35/Mb)and chromosome 3 had the lowest(469.65/Mb).The result showed that the frequency of microsatellite was not related to Tthe length of the chromosome DNA sequence and the frequency of microsatellite on it were uncorrelatedin which it was located.2.Genetic relationship analyzing of 10 markers with higher polymorphisms showed its cumulative discriminating power and probability of parental exclusion were higher than 0.99.10 high polymorphism microsatellite markers were screened by the parent population.By analyzing the genetic relationship between parents and four offspring,it was concluded that t The female parents of B1,B2,C1 and C2 were A-3,A-117,A-3 and A-117,respectively.The average allele was 8.3,the average polymorphism information content was 0.68~0.72.,and the cumulative parental exclusion rate was above 99%.Matching schemes based on genetic distances amongparents are beneficial to avoid genetic diversity loss,stress tolerance loss,growth slowdown and individual miniaturization due to inbreeding.In this study,we found that W,X,P,T,S populations with genetic distances between 0.54 and 0.60 were better as breeding parents for breeding.3.In this study,W,X,P,T and S populations were selected as parents of breeding group.For green arowan,Osteoglossum bicirrhosum,Heterotis niloticus,Osteoglossum ferreiai,red arowana,Arapaima gigas,Scleropages jardini,Scleropages Leichardti.The results of microsatellite marker analysis showed that the effective amplification ratios of green arowana,Osteoglossum bicirrhosum,Heterotis niloticus,Osteoglossum ferreiai,red arowana,Arapaima gigas,Scleropages jardini,Scleropages leichardti were78%,34%,16%,34%,82%,20%,68% and 32%,respectively.The This studyresearch reveals that us,ing microsatellite markers to solve the intraspecific relationship of gold arowanagolden dragon fish is feasible,which layings a foundation for the study ofestablishment and understanding of Osteoglossumgenetic diversity of the family of Scleropages formosus family.and The research the transferbility of the microsatellite markers.provides help for the further study on genetic relationship and genetic diversity analysis in golden arowana,and the transferbility of the microsatellite markers. |
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