Parentace Identification Using Microsatellite DNA And The Research On Epigenetic Regulation Under High Temperature Stress In Apostichopus Japonicas

1Parentace identification using microsatellite DNA in artificialbreeding of Apostichopus japonicusPaternity test was earliest used in judicial identification, with the development ofits technology, now is widely used in zoology, botany and ecology, etc. In geneticbreeding of aquatic animals, with the continuous development of modernbiotechnology, paternity test provide technical support for breeding and the pedigreetrack, application of paternity test technology to identify individual identity and makepedigree information clear has very important practical significance.In the present study,8microsatellite loci were adopted for parentageidentification of240offspring and total14parents in artificial breeding group ofApostichopus japonicus. The results showed that all the8microsatellite loci hadgenetic polymorphisms, the total number of alleles was53, and the mean number ofalleles was6.63.The observed heterozygosity ranged from0.730to0.902, and PICvalues were0.696-0.836. Analyzing by CERVUS3.0, the calculate exclusionprobabilities of microsatellite loci were0.9934,0.9997and0.99999.Based onElimination method and judged by the genotypes of parents and offspring,37individuals had one parents and194individuals had two or more candidate parents.Based on Likelihood method, there were198individuals found the most likeparentsat80percent confidence level. Parental Identificationon the results of thecomplementary methods,229individuals found their parents with95%success rate. The results can provide technical support and theoretical basis for parentageidentification in the breeding practice of Apostichopus japonicus.2Epigenetic regulation of related gene expression under hightemperature stressWhen organisms under adversity stress, the change of epigenetic modificationscan be induced in the body, they regulate the expression of stress response genes tochange the organism’s physiological state, ultimately achieve the goal of adapt to theenvironment. Epigenetic changes plays an important role in the organisms respondingto adversity stress, it is hoped that through the study of the choice of these phenotypicvariation in order to achieve improved biological targets. In a word, the further studyof epigenetic will help to further reveal its role in the biological art, for the use ofepigenetics for breeding to lay the solid theoretical basis.This study use the real-time fluorescent quantitative PCR, and through therelative quantitative2-△△CTmethod, housekeeping genes β-actin as internal calibration.Compared the expression of the four epigenetic regulation in the Apostichopusjaponicus breathing tree, bowel, body fluids and muscle tissue. The Apostichopusjaponicus under25℃as the control group, its aim gene expression as a benchmark1,the relative expression quantity of different genes under different temperature stressshow:in the body fluids, the expression of DNMT1was not significant increased at18℃,increased significantly at18℃to21℃, the risen is not obvious while21℃to24℃,gain the maximum expression3.26at30℃, the general expression isincrease. the expression quantity of MLL5and HDAC3genes is outstanding at24℃,the expression quantity is slightly higher as the temperature increases, the expressionquantity respectively gain the maximum expression of2.90and3.19at30℃. Overall,the expression quantity of all genes are increased as the temperature increase,up-regulation expression trend are obviously, highest expression reached at30℃.The data display, the expression of every genes in each organization is different,expression in a fluid is the highest, followed is bowel, respiratory tree, the lowest is muscle. The temperature stress conditions of30℃,detect the expression of3genes at0,3,6,6,9,12,15h, the expression quantity at0h as a benchmark, Result is shown,expression quantity of three genes are raising, in0-9h, the expression of upward trendis obvious with the extension of stress time; with the risen of temperature, theexpression quantity of DNMT1and HDAC3has a downward trend when reached thehighest expression quantity at9h, but the overall expression is still raising. Theexpression of genes in body fluids is higher than the other organizations.