Study On RAPD Analysis Of 38 Germplasm Resources And Molecular Marker Of Several Important Agronomic Characters In Sugarbeet

High sugar, high output or haloduric character of sugarbeet takes as a significant value which weighs fruit output, in search of molecular markers which tightly linked to high sugar, high output or haloduric gene, as it has significance of sugarbeet breeding .In this research, molecular markers linked to high sugar, high output or haloduric character of sugarbeet were studied with RAPD and SSR technology in 112, 154 or 174 F2 hybrided population from high-sugar and low-sugar sugarbeet, L242 and L236 or haloduric×sensitive salt sugarbeet. The main results were as follows:1 Sugarbeet total DNA was extracted with four different methods CTAB,SDS,improved CTAB and improved SDS. The DNA was analyzed with concentration test , ratios of D260 and D280. The result show that: improved CTAB method DNA content yield higher than others. Therefore, improved CTAB method was a best and optimum way for sugarbeet DNA extraction. A procedure of improved CTAB had been modified. 764μg of sugarbeet DNA was obtain from 1.0g sugarbeet spawn under the following conditions: 5.0 mL cell extraction solution, 0.8mL ethanol dosage. The rate between isopropanol and the test solution was 0.6 volume.2 The L25 (56) orthogonal diagram was applied to study the effects of different concentrations of Taq polymerase, Mg²⁺ , dNTPs, primer and DNA on sugarbeet. RAPD-PCR results and to obtain a suitable PCR reaction system. The results showed that the orthogonal design could be used to optimize the RAPD-PCR system. A suitable RAPD reaction system for Sugarbeet was developed, i.e.l×Taq polymerase reaction buffer, 1.0mmol/L Mg²⁺, 1U Taq DNA polymerase, 0.25mmol/L dNTPs, 0.4μmol/L primer and 25ng genomic DNA template with total 25μL reaction solution. It will have high resolution in polyacylamide gels electrophoresis.3 In this study, we optimizatied SSR condition on Sugarbeet. In 20μL PCR reaction, 2.0mmol/L Mg²⁺; 0.20 mmol/L dNTPs; l.5U TaqDNA polymerase, 1.5μmol/L primer; 60ng/μL DNA are the best.4 The 38 sugarbeet varieties of different sources were studied in the first part of the paper. The analysis based on Randomly Amplified Polymorphic DNAs analysis. Result shows the sugarbeets are polymorphic. The genetic resource is abundance. The relationship of haloduric and sensitive salt or L242 and L236 sugarbeet parents is similar with the same resourse, but the relationship of high-sugar and low-sugar sugarbeet parents is not.5 There are 64 RAPD primers and 2 SSR pairs primer that may recognize the material of high-sugar and low-sugar sugarbeet ; There are 60 RAPD primers and 4 SSR pairs primer that may recognize the material of L242 and L236 sugarbeet ; There are 20 RAPD primers and 5 SSR pairs primer that may recognize the material of haloduric and sensitive salt sugarbeet.