Establishment SSR Fingerprinting And Evaluation Genetic Diversity Of Maize (Zea May L.)

100 SSR primers screened were analyzed and finally 20 primers fitting for construct maize inbred line and hybrid DNA fingerprinting map were confirmed. And SSR technology system fitted for maize seed DNA was established. The genetic diversity of twelve or twenty-four or forty-two maize inbred lines were researched. The main results were as follows:1. SSR technology system fitted for maize seed DNA was established: The genomic DNA of maize was extracted by a modified SDS method(The modified extraction buffer contained 100 mmol/L Tris-HCl (pH 8.0);50 mmol/LEDTA, 100mmo1/L NaCl, 1.5% SDS, and DNA was purified by chloroform.).The gained genomic DNA was suitable for SSR analysis. Modified SSR reaction system(20ul reaction system contained2.0mmol/LMg2+,0.1mmol/LdNTPs,0.25μmol/Lprimer,0.5UTaqE,10×PCR buffer 2.0μl and 40ng DNA). The outputs of PCR were electrophoresed on 10% non-denaturing acrylamide in 1×TBE buffer at constant voltage (140V or 120V) for 3.5-4.5h., and then reveal the bands through modified silver stained method.2. Eight pairs of maize SSR primers were used for building multiplex-PCR according to their amplified bands' size range. Under the same conditions as single PCR reaction, 28 Double-PCR appeared three different amplification behaviors, of which 12 were amplified normally. This screened normally-amplified multiplex-PCR could be applied in maize DNA fingerprinting database.3. Primer select:Primers were excluded from the study if banding patterns were difficult to score accurately on gels or if the primers failed to amplify consistently in all 12 inbred lines. A final set of 20 SSR primers was chosen for further analysis, and finally 12 mainly primers fitting for establishing maize inbred line and hybrid DNA fingerprinting map were...