Studies On Rapid Testing Genetic Purity Of Sunflower Seeds With Indoor Method

Seed is an important agricultural product material, and seed genetic purity is the key factor on playing the potential of yield increasing for improved variety, therefore, seed genetic purity identification is of important significance to ensure seed quality. Genetic purity testing is an important part of quality control for production of sunflower hybrid seeds. The traditional determine methods play an important role in testing the genetic purity of sunflower seed, but due to some limitations, resulting in these methods can not meet the requirements of rapid and accurate detection of seed genetic purity. So, there is an urgent need to develop a simple and quick way to determine the genetic purity of sunflower seeds. This study is to develop a rapid indoor way to determine the genetic purity of sunflower seeds on the basis of the AU-PAGE method to measure the maize seeds genetic purity and rapid test genetic purity of maize seeds with molecular technique, through studying the protein electrophoresis method and SSR molecular technique to test the genetic purity of sunflower seeds. The main results were as follows:1. We established the sunflower seed protein electrophoresis detection technology.Through the study of sunflower seed storage protein composition, the results showed that the most abundant proteins in sunflower seeds were water soluble proteins, followed by salt soluble proteins, while other proteins were relatively few. Therefore, the purity testing could be conducted based on the polymorphisms of water-soluble and salt-soluble proteins. Through optimizing the sunflower protein extraction buffer, we showed to increase the clarity of the gels by adding appropriate amount of SDS in the protein extraction buffer. The reduced background staining of the gels improved the band readability and thereby increased the protein polymorphism. Optimized protein extraction buffer parameters: 18% of urea, 12%acetic acid, 3% TEMED, 0.15%-0.2% SDS, 0.05% methyl green. The nine parental lines as well as six hybrids could be readily distinguished each other with optimized AU-PAGE method to identify six hybrids and nine parents. With a new naming method for six sunflower hybrids and nine parents established a protein fingerprint database, so any parent composition could find the specific protein band.2. Through the optimized of DNA rapid extraction method, we established the rapid DNA extraction method from half a sunflower seed for PCR. We have studied the effect ofconcentration and volume ratio of extracting solution and extracting time and amount of DNA templates on PCR products. The parameters of DNA rapid extraction method were confirmed.100 high polymorphism and good specificity primer were screened from 215 pairs of SSR primers and was established a database using 11 sunflower parents as experimental material.Rapid DNA extraction method from half a sunflower seed for PCR and universal multiplex PCR technique are combined into a rapid testing genetic purity of sunflower seeds with molecular technique.3. Protein electrophoresis and SSR rapid molecular test results were compared with examination of plants on field plots. The results show that the purity of examination of indoor is lower than that of plants on field plots. The accuracy for indoor determination method to examination hybrid purity is higher. Parents could not only be detected, but also hybrid strains caused by parents impure could be identified. Therefore, the method was suitable for sunflower seed genetic purity testing.