Study On The Construction Of DNA Fingerprint Database Of Japonica Rice In Heilongjang Province By SSR Technology

Heilongjiang japonica rice is high-quality with enormous commercial value. But rice adulteration is serious. SSR molecular markers is screened to implement this experiment, with 61 samples that planted broadly in Heilongjiang Province. 19 pairs of core SSR primers are selected to construct DNA fingerprint then research genetic differences in these Japonica rice varieties. Hoping to identify them accurately and effectively identification from the molecular level and solve adulteration in the rice market and also providing the theoretical basis for the protection of genetic diversity of rice in Heilongjiang Province and cultivation of excellent rice varieties. The results are showed as follows:1.Optimization of DNA extraction method from rice. Optimization is carried out on the basis of improved CTAB method and the results show that DNA extraction concentration can meet the requirements of the SSR experiment after the improvement.2.Optimization of SSR technology system. The SSR technology system was optimized, including SSR-PCR system, denaturing polyacrylamide gel electrophoresis system and silver staining system. The experimental results show that the optimized SSR technology system can get better experimental results, which is beneficial to the statistics and analysis of the SSR band information in the next step.3.Screening of core SSR primers and construction of DNA fingerprinting. Using the optimized experimental conditions to conduct SSR markers experiment. 19 core SSR primers were screened to construct the DNA fingerprint database of 61 copies of rice in Heilongjiang Province with good amplification effect and clear band, good polymorphism.4.Polymorphism analysis of SSR primers was executed by the DNA fingerprint info rmation gained,results show that: the 19 pairs of core SSR primers amplified 71 alle les in 61 rice materials. Cluster analysis results based on SSR markers showed that t he 61 samples tested rice can be divided into two groups,the first category contains 51 materials, is classified into several small groups, the genetic relationship of all s amples can be distinguished. The results of genetic diversity analysis showed that t he core SSR primers can provide a little amount of polymorphism information and t he genetic diversity between the rice materials is not rich enough.