Collection Of Germplasm Resources And Breeds Conservation Of Embryo Skin Fibroblast Cell Of Goose In Jilin Province

Zi Goose and Huoyan Goose are excellent local breeds in Jilin Province with the characteristics of cold tolerance,crude feeding tolerance and high egg production.These breeds thereby are strong candidates for female breeds in conventional cross breeding.Therefore,In recent years,blood purity has been reduced due to commercial disordered hybridization and the lack of protection awareness of the industry and farmers,which has greatly destroyed the population structure of the original varieties.targeted conservation is extremely urgent.This work combined with the geographical distribution characteristics of the varieties in Jilin Province,to conduct the analyses of the geographical conditions such as weather and light in the regions where the breeds live,further to carry out the identification of the appearance characteristics of local breeds in several regions and the collection of eggs.The egg weight and egg shape index were measured.In the work of cell-level genetic resources protection,Zi Goose eggs reached the standard were hatched.Embryonic skin was collected at E12 for cell culture using pancreatic enzyme method after purification.Fibroblast cell line of Zi Goose was established via the original generation,subculture,cryopreservation and resuscitation.Biological activity of the cultured cells was detected.We next conduct the work of cryopreservation and genetic resources protection for subculture cells.The standard of the cell line establishment was reached by investigating cell resuscitation and cell viability.Which indicated that the goose embryonic skin fibroblast cell line was established successfully.The main results are as follows:1.Data collection of germplasm status local goose breeds and main exotic breeds in Jilin Province.The main three raising goose regions were divided following the zoning principle: western prairie region(Songyuan City and Baicheng City),central agricultural region(Siping City and Jilin City)and eastern mountain region(Liaoyuan City and Tonghua City)for white goose distribution.We visited to local industries and farmers,and collected 5 breeds breeder eggs from 22 counties.The egg weight,egg shape index and fertilization rate were measured.In the comparison analysis of the egg weight and egg shape index among the three regions,egg weight of Zi Goose was significantly different among the three regions(P<0.05).Egg weight of western prairie white goose region typically in Songyuan City was the lowest,and significantly different with central agricultural white goose region and eastern mountain white goose region(P<0.05).Egg shape index was significantly different among breeds in some regions(P<0.05).The number of breeder eggs,reaching the optimal range of egg shape index(1.349～1.449),of Zi Goose in western prairie white goose region was the most.There was a negative correlation between fertilization rate and temperature in all regions(P<0.05).In the comparison analysis of the egg weight and egg shape index in Huoyan Goose,Jilin City of central agricultural white goose region was significantly higher than Tonghua City of eastern mountain region(P<0.05).The number of breeder eggs,reaching the optimal range of egg shape index(1.350～1.490),of Huoyan Goose in eastern mountain white goose region was the most.In the comparison of exotic breeds,there was no significant difference in egg shape index between Carlos Goose,Hortobágy Goose and Zhedong White Goose(P>0.05).Egg weight of Carlos Goose was significantly lower than that of Hortobágy Goose(P<0.05).2.Establishment of Zi Goose embryonic skin fibroblast cell conservation.Immunofluorescence identification showed that the cultured cells were typical fibroblasts with fusiform or star-shaped flat morphology and radiological distribution.The growth curve of goose embryonic fibroblast cells showed an "S" shape.Cells experienced three stages of incubation,exponential proliferation and flat top.Cell proliferation was normal and in accordance with the law of in vitro growth.The results of cell viability measurement showed that the cell viability of resuscitation was lower than that before cryopreservation in the third-generation cells,but the viability was still above 90%.Indicating that the cells might be damaged in the process of cryopreservation and resuscitation,but it met the requirements of cell line establishment.In the microbial detection for goose embryonic fibroblast cell line,the all results were negative without bacteria,fungi,viruses and mycoplasma contamination in the cell culture fluid.We carried out genetic resources protection using subculture cell cryopreservation.Standard of cell line establishment has been reached via assessing cell resuscitation and cell viability.A total of 242 samples of the primary,first,second and third generation cells were cryopreserved.To protect genetic resources,this work conducted data collection and measurement according to the characteristics of goose breeds resources distribution in Jilin Province,based on the investigation of breeds.Furthermore,fibroblast cell conservation of Zi Goose embryonic skin was established using cell culture and et al.methods for further exploration of cell-level genetic resources protection.This study might be contributed to Chinese goose genetic resources protection and theoretical basis.