Studies On The Genetic Relationship Of The Crested Ibis Using RAPD And Blood Protein Polymorphism

With the help of Ramdomly Amplified Polymorphic DNA (RAPD) markers and blood protein polymorphism, the genetic relationship among the 40 crested ibis' was identified and found suitable PCR amplified system. At the same time, it was explored in 4 blood protein locis of the crested ibis about Tf, Pa, Alb and Pre.The results showed:(1) The effect of RAPD is easily affected by many factors, such as the concentrations of Taq enzyme, primers, Mg2+ and the amount of template DNA. The ladder experiments showed: the best PCR amplified system of the crested ibis is as follows: Primer(10pmol/ul)lul, Taq enzyme(lU/ul)1.5ul, dNTP(2.5mM)2.0ul, Mg2+(25mM)2.5ul, teplate DNAlOng, TH2O17ul, and final volume is 25ul. The suitable RAPD-PCR amplified procedure is: preview heating at 94 ℃ for 30s; annealing at 36 ℃ for Imin; extending at 72℃ for 2min; store at 4℃ in the end.(2) Under the previously optimizing conditions, ten ideal primers:B-03, B-09,B-19,B-20,B-36 were screened from 40 random primers, which accounted for 12.5%. It was found in this study that 742 DNA fragments were amplified, including 582 polymorphic fragments, which accounted for 74.8%.(3) Accoring to the amplified results, the trees were conducted by "PiErxun" and "Ou's" method using SPSS software. From the trees, we can see the genetic relationship is very close between 19 and 32, 18and 26, 22 and 23, 5 and 31,and also find it is relatively distanter between 28 and 29, 39 and 36, 2 and 33, 38 and 12. The results obtained from the the trees of "PiErxun" and "Ou's" are similar, which distinctly reflected the genetic relationship among the 40 crested ibis.(4) The PAGE results indicated there were no polymorphic locis in Tt, Pa, Alb and Pre. RAPD results showed the average BSR of the 40 crested ibis' is 0.86, which revealed there still 14% genetic variability.