Study On The Method Of Rapid Determination Of The Karyotype Of Hybrid Buffalos And The Relationship Between Karyotype And Reproduction

Buffalo is divided into two types,swamp buffalo and river buffalo,the chromosome of swamp buffalo is 48 while the chromosome of river buffalo is 50. the number of chromosomes of F1 generation of two types buffalo hybrid is 49.The chromosome numbers of offspring of F1 generation and river type buffalo hybridize, or three kinds of different chromosome numbers of buffalo hybrid are 2n=48, 2n=49, 2n=50. The culture conditions and the subsequent steps wereoptimized,further improving the efficiency and accuracy of the experiment.we obtain the chromosome numbers of 128 hybrid buffalos. In order to provide theoretical basis for the development of milk buffalo breeding program,we carry on the correlation analysis between different chromosome and lactation performance, reproductive performance, body size. The operation of Peripheral lymphocyte culturing and chromosome karyotypes analysis are complicated. In order to accquire a simple, accurate and reliable chrosome karyotype analysis method, we verify the enzyme cutting site of PCR-RFLP via expanding the samples.The results are as following:1. Acquire 128 heads of hybrid buffalos chromosome karyotypes data, including 53 heads in 2n = 50(41.41 %), 46 heads in 2n =49( 35.93%), 29 heads in 2n =48(22.66 %)2. Analysis the factors of affecting the chromosome karyotypes, we found that 4 factors include the dose of heparin,permeability time, buffalo’s age,centrifuge speed. In winter, added amount of heparin for adult bovine and calf are 140μl(500U/m L) and 260μl(500U / ml). The optimized permeability time is about 40 minutes. The rotational speed is 800-1000(r/min), the centrifugal time is 8min.3. Three different karyotypes of reproductive performance in the first parity were compared.2n=49 breeding index far greater than the other two types of Buffalo. The first and second calving interval of 2n=49 was longer than that of the other two types Buffalo.4. Three different karyotypes of lactation performance of buffalo in the first parity were compared. The buffalo of 2n=50 in average milk volume peak of lactation and the first three months of the Milk volume was significantly higher than the other two kinds of nuclear buffalo of average milk volume peak of lactation and the first three months of the Milk volume.In the first parity the Lactation days of 2n=50 is much longer than the Lactation days of other two types buffalos.5. The number of different karyotypes and the corresponding body size data were compared. Found several indicators of significant differences in the rump length, body length, and nipple spacing, the abdominal circumference.6. Using the technology of PCR-RFLP found that the accuracy site of RBP3-4+ identification of 2n=50 buffalo is 85.19%.The accuracy of RBP3-4+ identification of 2n=49 buffalo site is 64.2%. The accuracy of RBP3-2 identification of 2n=48 buffalo site is 80.59%...