Using SRAP Marker To Construct The DNA Fingerprint Of Excellent Varieties Of Peaonia Suffruticosa In Heze City

33 different varieties of Peaonia suffruticosa in Heze were chosen as the reaction materials. DNA from those materials was extracted by CTAB method. Based on the optimal reaction suitable for SRAP-PCR amplification, SRAP marker was used to construct the DNA fingerprint of those different varieties.Based on CTAB method,the efficiency of different DNA extraction methods was compared. Finally, an appropriate way to extract DNA from Peaonia suffruticosa was determined. It was CTAB cracking, then extracted once by chloroform-isoamyl.Orthogonal experimental design was applied to establish the optimal reaction system for SRAP–PCR which was 25μL reaction volume. It included 0.5U Taq DNA polymerase, 2.0mmol/L Mg2+, 50ng template DNA,0.2 mmol/L dNTPs, 0.30μmol/L primer. Amplification program was 5min at 94℃; 1min at 94℃, 1 min at 35℃, 1.5min at 72℃, 5 cycles; 1min at 94℃, 1min at 50℃, 1.5min at 72℃and 35 cycles; then 10min at 72℃and stored at 4℃.There were 16 primer combinations selected from 648 primer combinations, which were applied in genetic diversity and systematic relationships.The number of amplified fragments per primer combination was 20.4 while the number of those was 11.2. The percentage of polymorphic fragments was 54.2%. Results showed that SRAP marker was one of the most efficient, economic and dependable ways of molecule marker.Primers combination Em23-Me15 could distinguish those 33 different verities of Paeonia suffruticosa. Rich and clear DNA fingerprinting was obtained. The range of SRAP-PCR products was between 100bp to 1000bp.Genetic similarity coefficients were calculated and dendrograms were constructed by the unweighted pair group method of arithmetic average (UPGMA) under the software NTSYS-pc version 2.10. The results showed that anemone type and crown type had a close relationship. There was a trend of single flower and tower flower belonging to different cluster. The results were inconsistent with the taxonomy based on the flower type, which might be related to long-term introduction, selection and crossbreeding. In all, all sorts of genetic markers should be comprehensively used in cultivar classification of Paeonia suffruticosa.So it's important to set up the standard DNA fingerprint of the variety of Paeonia suffruticosa under SRAP reaction system. The research achieved valuable experience on guiding the new varieties of the Paeonia suffruticosa of international cultivar registration and instruction breeding.