| Xinjiang is the province of China ,where sheep breeds is the most largest and has complex genetic quality, it has bred a number of characteristics local varieties that is a valuable asset for development and utilization of the world's animal resources in the long-term production practice. Kazakhstan sheep has excellent traits such as large body , prematurity, resistance, and adaptability;Altay sheep is a good group of Kazakhstan sheep that have much feature for example large body and fast growth of lamb and fast fat and high-fat meat production; Bashibai sheep also has many features such as prematurity,resistant coarse feed,strong adaptability and brown-red hair and multi-pile and dead hair less dry and so on. These local varieties is the important species that is used to protect the biological diversity,and it is to achieve the basis of sustainable development of sheep-raising in China. In this study, for the first time use AFLP markers to clarify polymorphism of Xinjiang breeds resources among Kazak sheep, Altay sheep,and Bashibai sheep from the molecular level,it will provide a scientific basis for conservation and rational forthputting of the local species.Using of the AFLP markers, this experiment has accomplished genetic analysis to Kazakh sheep, Bashibai sheep Altay sheep in Xinjiang,has counted the polymorphic bands that each primer combinations detected in all varieties;has calculated genetic similarity coefficient genetic distance of three local varieties sheep ,and based on that, constructed the UPGMA cluster diagram. The results are as follows:1. Comparison of the boiling water bath,Miller salting-out method,CTAB method and phenol extraction improved four kinds of sheep genomic DNA extraction method,which extracted in boiling water bath for purity and concentration of DNA was significantly lower than the other three kinds of methods. Phenol extraction improved with Miller salting-out method,CTAB extraction method of DNA compared to the purity and concentration of the product of a significant difference (P<0.05),suitable for AFLP molecular markers of DNA Experimental request template.2. This study had gone through a number of repeatedly screening to a total of 64 MseI and EcoRI primer combinations, in which obtained 11 primer combinations with more bands and higher percentage of polymorphism, richer banding pattern,clearer strips (E01/M01,E01/M07, E02/M03, E04/M01, E05/M05, E05/M08, E06/M02, E06/M04, E06/M05, E07/M03, E07/M08). A total of 527 strips in the three local sheep varieties, on average,47.91 strips were amplified per primers combination, the scope of the changes is the 34 to 61,including 46 polymorphic markers,with a polymorphic frequency of 8.73%, on average 4.18 polymorphic markers polymorphic markers were amplified per primers combination.3.The 11 selective primer combinations by which more polymorphic markers (showed in brackets)were obtained involved in E01/M02 ( 7 ) ,E07/M08(6),E06/M02 ( 5 ) ,E06/M04(5),E07/M03(4),E02/M02(4),E05/M08(4).4. According to the calculation result of the 11 AFLP primer combination,it hascalculated genetic similarity coefficient genetic distance of three local sheep varieties, in which the biggest genetic similarity coefficient is Kazakhstan and Altay sheep(0.699), the smallest genetic similarity coefficient is Kazakhstan sheep and Bashibai sheep( 0.592), and the genetic similarity coefficient of Altay sheep and Bashibai sheep is 0.620.5.Using UPGMA method of NTSYSpc software to carry on cluster analysist to the three local sheep of Xinjiang,and constructed the UPGMA cluster diagram. |
PDF Full Text Request