Study Of The Difference On DNA Methylation Of Putian Black Pig

In this study, the methylation patterns of different tissues in different developmental stages of Putian black pigs were detected and analyzed by MSAP technique, and adopted BSP technique to analyze methylation status of H-FABP gene promoter in different tissues of Putian Black pigs, Duroc pigs and the hybrids of Duroc pigs with Putian pigs(Dupu pigs),the research will provide theoretical basis for molecular breeding of pig breeding work and fat deposition in pigs.1. To detect the genome methylation status of Putian black pig at different stages of growth and development, we selected 16 pairs of primer combinations to use MSAP technology, The main results obtained were as follows:(1) Analysis of genome methylation status of Putian black pig during the age of 1 day, a total of 12324 bands were amplified, of which 2510 were hemimethylation bands,3557 were full methylation bands, of which 6257 were non methylated bands.In the level of hemimethylation, the level of the liver was significantly higher than that of the heart, muscle and tail (P<0.05). About the full methylation level, the level of the heart was significantly higher than that of liver, muscle, fat, ear and tail respectively (P<0.05).In the total methylation level, the level of heart was significantly higher than that of liver, muscle, fat, ears and tail, respectively (P 0.05); the level of tail was significantly higher than that of muscle, fat and ear, respectively (P< 0.05), significantly lower than the heart and liver (P< 0.05).(2) Analysis of genome methylation status of Putian black pig during the age of 25days, a total of 11505 bands were amplified, of which 1997 were hemimethylation bands,3740 were full methylation bands,5768 were non methylated bands. In the level of hemimethylation, the level of heart was significantly higher than muscle (P< 0.05); the level of liver was significantly lower than fat (P< 0.05); the level of muscle was significantly lower than heart, fat and ear, respectively{P< 0.05). The difference was not significant between the heart, liver, muscle, fat, ear and tail in the full methylation.In the total methylation level, the level of the heart was significantly higher than muscle (P<0.05), was significantly lower than that of fat and ear (P< 0.05); the total methylation level of muscle were significantly lower than heart, fat, ear and tail, respectively (P<0.05).(3)Analysis of genome methylation status of Putian black pig during the age of 210days, a total of 11850 bands were amplified, of which 2294 were hemimethylation bands,2867 were full methylation bands,6689 were non methylated bands. In the level of hemimethylation, the level of hemimethylation of heart was significantly lower than that of fat, ears and tail, respectively (P<0.05); the level of liver was significantly lower than that of fat, ears and tail, respectively (P<0.05); the level of muscle was significantly lower than that of fat and ear (P< 0.05). The difference was not significant between the heart, liver, muscle, fat, ear and tail in the full methylation level. In the total methylation level, the total methylation level of heart was significantly lower than tail (P< 0.05); the level of liver was significantly lower than that of fat, ear and tail, respectively (P< 0.05); the level of muscle was significantly lower than fat and tail (P< 0.05).2. The full methylation levels were higher than hemimethylation levels in heart, liver, muscle, fat, ear and tail of Putian black pigs in different developmental stages.3. The average total methylation levels at 1 day,25days and 210days of Putian black pigs were 50.67%,49.8%,43.55%, which were along with the age growth decreased.4. The hemimethylation, full methylation and total methylation levels in different gender of Putian Black pigs were not significant (P> 0.05), the hemimethylation levels were less than the full methylation in same gender, the total methylation level of males in different developmental stages were slightly higher than that of females.5. This study detected by BSP method to analyze that the methylation levels of H-FABP gene promoter in heart, muscle and ear of Putian Black pigs, Dupu pigs and Duroc pigs at 210 days, the methylation levels exhibited lower levels.6. Differences of methylation levels of H-FABP gene promoter in heart, muscle and ear of different pig breeds were not significant. The differences of methylation levels of H-FABP gene promoter in the same tissue of different pig breeds were not significant.7. Analysis of the methylation status of CpG site in H-FABP gene promoter inheart, muscle and ear of Putian Black pigs, Dupu pigs and Duroc pigs, a total of 35 CpG loci were detected ineach tissue. There were 3 high levels of methylation of CpG loci that were detected in three tissues of three pig breeds, and they had the same sequence position.