Drought-resistance And Sifting Molecule Mark Of RAPD By Sugar Beet Ion Impregnating Into Mutagenic High-sugar Mutant

Xinjiang is one of major sugar beet producing areas in China, and leading in yield. The study was under taken to identify high-sugar beet material obtained by ion impregnating with mutagerlie technology and another material which has not been mutagenized by molecular marker, and obtained linkage map in high-sugar beet primarily. The studies Strengthening choice and identification selection of target gene, increasing new material and shorten life of breeding.In our experiment,DNA was extracted and RAPD was analyzed by using sugar beet sugar mutant S10 obtained by impregnating ion with mutagerlie technology and F104 which has not been mutagenized . Drought-resistance of S10 was anysised.The main results are as follows:1.Many DNA extraction methods were studied with aim to find a simple and rapid method to identify sugar beet resources and to do other molecular biological researches. The results showed that the modified SDS method was suitable to extract sugar beet DNA based on the purify, the yield and effect of DNA. The products (bands) were clear and no degradation in the gelelectrophoresis. The value of OD260/OD280 were between 1.6².0 detected by Ultraspec U/V spectrophotometer. This method could get good DNA and was qualified to apply to RAPD-PCR about high-sugar beet.2.The different PCR program and the ingredients in RAPD reaction system such as renature , cycles ,Taq DNA polymerase, Mg²⁺, dNTPs, DNA and primers were screened by different concentrations. Suitable reaction system and program of the RAPD analysis for sugar beet DNA was settled. The following was the 25ul-reaction system with 60ng template DNA, 10×Buffer2.5ul;2.5mmol/L Mg²⁺;0.2mmol/L dNDTPs;2.5UTaq DNA polymerase;0.10mmol/Lprimer; The program was that pre-denature at 95℃for 5 min, then 94℃for 1min, 36℃for 1min, 72℃for2min, for 45 cycles,finally extended at 72℃for 5min.3.The S10 and F104 increased by randomly selecting 800 primers of RAPD.The result indicated that there was difference on DNA level between S10 and F104. There was difference in S10 and F104 of 19 primers which had randomly sifted from 800 ones.The number of the primers with difference was up to 2.3% of the total. It indicated that S10 was very much similar to F104.4. Linkage map had been set in sugar beet. 5 linkage maps consisted 15 markers are existed. Total genetic distance is 202.4cM.5. Compaired S10 with other materials from content of chlorophyll content of proline content of protein , drought-resistance of S10 was poor primarily.