| Hexagrammos otakii(Fat greenling)belonging to family Hexagrammidae and genus Hexagrammos,mainly distributed in North West Pacific.H.otakii is an important kind of economic fish species in East Asia.In this research,we analyzed polymorphism difference between geographical population and difference between wild and cultured groups by mitochondrion marker.Using 2b-RAD method combined with high-throughput sequencing to develop S SR and SNP markers which associated with growth and development.Correlational analyses between SNP and growth traits.Nucleotide sequences of mitochondrial DNA cytb and D-loop from four different population of Hexagrammos otakii were amplified by using PCR technique,365 bp and 387 bp nucleotide sequences were obtained to study on the genetic diversity of Hexagrammos otakii,respectively.The result showed that the total variable sites,number of haplotypes(h),haplotype diversity(Hd),nucleotide diversity(?)and mean pairwise nucleotide differences(k)of four populations based on cCytb were 10,7.75,0.739,0.007 3,2.656 respectively;based on D-loop were 18.25,12.75,0.846,0.012 1,4.673 respectively.Diversity from high to low was Zhoushan,Dalian,Langyatai,Jimo(cultured population).Diversity in cultured population is lower than that in wild population.Fst value based on the analysis of molecular variance(AMOVA)of wild populations' Cytb and D-loop was 0.318 6 and 0.271 4,respectively.It shows that the within population variation is the mainly variation.The NJ molecular phylogenetic trees based on Kimura 2-parameter model showed that three different geographic stocks have no apparent Geographic subdivision.The sequencing of H.otakii mined 4,366,696 reads,2,183,348,000 bp,Q20 is 87.6%,Q30 is 83.11%.We isolated 20 novel microsatellites of Hexagrammos otakiii(Fat greenling)which associated with growth by using 2b-RAD sequencing method.The characteristics of 20 microsatellite loci were amplified in 105 Hexagrammos otakii individuals which came from four different groups and tested by Capillary electrophoresis.The number of allele of the 20 microsatellite loci ranged from 8 to 26 with an average of 19.95.PIC value ranged from 0.1659 to 0.9227 with an average of 0.7555.Observed and expected heterozygosity varied from 0.0667 to 0.8571 and from 0.1710 to 0.9314,respectively.Highly polymorphic characteristics were observed in each microsatellite loci of the study except H06.Further research showed that 10 microsatellite loci had transferability in Hexagrammos agrammus.In this study,20 polymorphic microsatellite loci were provided have sufficient level of genetic diversity to investigate the fine-scale population structure,genetic relationship and evaluate the artificial reproduction strategy for further study in Hexagrammos otakii.In this study,158,733 SNP were found,after screen,21 SNP markers were developed.17 of the 20 have polymorphism,HO ranged from 0.0687 to 0.5115 with average 0.2666;HE ranged from 0.0666 to 0.6815 with average value 0.3005.HOS2,HOS6,HOS14 deviated from HWE,minor allele frequency of HOS19 was lower than 0.05.Correlational analyses showed that HOS17 has significant influence in weight growth,HOS13 and HOS21 have significant influence in body length growth,HOS 18 has significant influence in weight and body length growth. |
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