Research And Application On Standard DNA Fingerprinting Technology Of Sweet, Waxy Maize Varieties

In this study, 165 varieties of sweet, waxy maize were conducted to investigate the impact of sweet, waxy maize DNA that was drawn with different methods on fingerprint atlas. Based on fingerprinting techniques using in common maize, 10 pairs of core primers were selected to construct standard DNA fingerprinting database of sweet, waxy maize varieties. The purity and authenticity qualification to the varieties of sweet, waxy maize was identified for the first time. The main results were as follows:1. SSR technology system fitted for sweet, waxy maize varieties DNA was established. Including reaction of DNA extraction (DNA of seed and leaf), PCR amplification system (20μL solution including: 11.4μL ddH2O, 2μL 1×Buffer, 2μL 2.5mmol/L MgCl2, 0.4μL 0.15mmol/L dNTP, 2μL 0.25μmol/L SSR primers, 0.2μL 1U TaqDNA polymerase, 2μL DNA template), denaturing polyacrylamide gel electrophoresis, silver staining method improved color, etc..2. Primer select: the core primers from 40 pairs of SSR primers for common maize were selected, which gave clear, stable, repeated, polymorphic amplification profiles, considering the ease of Statistics and high or low repetitive primers, and identified bnlg439, bnlg125, umc2105, phi072, umc1705, bnlg161, bnlg1792, phi080, phi065, bnlg1450 a total of 10 primer pairs as a sweet, waxy maize variety identification and fingerprinting database to build the core primers.3. The database of DNA fingerprintings of sweet, waxy maize was established. We set up 62 varieties of sweet maize and waxy maize DNA fingerprinting database through consistency and authenticity of the maize testing, using the sweet, waxy maize SSR technology system.4. SSR molecular marker technology was feasible to test the purity of sweet, waxy maize variety. Analysis of test results: a total of 24 poor or less consistency of sweet, waxy maize were stopped to test in 2006; a total of 9 poor or less consistency of sweet, waxy maize were stopped to test in 2007.5. SSR molecular marker technology was also feasible to identify the authenticity of sweet, waxy maize varieties. Identification of the authenticity of the results: There were no similar or identical species in regional trial in 2006. In regional trial, SN528 and SN5660 were same varieties; SN528 and NY1 were similar varieties, so NY1 and SN5660, the rest of varieties were different varieties in 2007.