Establish Of Rapd Fingerprints In Eucommia Ulmoides Olive.

Eucommia is a multipurpose tree species belonging to the order of eucommia family, as an extremely important basic work in the exploitation of resource, the breeding of Eucommia gets more attention than before. Recently, more and more cultivars (clones) which have good character was bred, and cultivated. But there is a little difference in characteristic between cultivars (clones), and make identifying more difficultly when meet a requirement, In order to provide a dependable technology for the identifying species, and study the difference in heredity, we established the fingerprinting of 14 cultivars (clones) by the RAPD technology and analyzed it. The main results are as below:1.Establish a improved method used to extract total DNA from the leaves of eucommia,tested and compared the reaction system and the cycle program. The total DNA extracted by the improved CTAB method, can get a good RAPD fingerprints. The reaction system used in this study were as follows: 10×buffer 2.5μL, Taq DNA polymerase 1U, MgCl2(25mmol/L) 2.0μL, dNTP(2.5mmol/L) 2.0μL, prime (5μmol/L)2.0μL, template DNA 2mg/L in 25μL reaction system. The PCR reaction program was: predenature at 94℃for 5 min, then 94℃30s, 37℃30s, 72℃90s, 35 cycles, and finally extended at 72℃for 10min, keep the PCR products at 4℃; 2.32 primers which has amplified and polymorphic bands had been selected from 500 random primers, the 32 primers are amplified for 14 cultivars, there are 191 amplified primers and 117 primers showing abundant polymorphisms, accounting for 61.26% .DNA bands are between 4 and 9 according to amplifying of different primers, the average is 5.96;3.The RAPD fingerprinting is constructed by the primers S465, S480 and S486. which can provides a references for cultivars(clones) identifying;4.14 cultivars(clones) are analyzed, including coefficient analysis and cluster analysis, and established the genetic dendrogram.the range of coefficient is between 0.56 and 0.93, average is 0.82,when coefficient was 0.82,the cultivars(clones) could be classified into five categories;...