Rapid Identification of Onion Hybrid Purity by Molecular Markers Based on Dried Seed DNA
In order to find a method for extracting high-quality DNA quickly from onion seeds and satisfy the requirements for accurate identification of the purity of a large number of onion hybrids,the DNA quality of onion seeds extracted by the TransGen PlantZol kit,TIANGEN rapid plant genomic DNA extraction system,TPS method and CTAB method were compared firstly in the study,and then the optimal extraction method was selected.The seeds with different germination days and the number of seeds for extracting effective DNA were also optimized and screened.Then,SCAR molecular markers were used to identify the purity of onion hybrid seeds to further verify the extraction quality of onion seed DNA.The results showed that the TIANGEN rapid plant genomic DNA extraction system failed to obtain the total genomic DNA of dried onion seed.The bands of DNA extracted by CTAB method were neat and consistent,single and bright,no tailing and degradation,and no obvious RNA and protein pollution.The DNA extracted from one dried onion seed and the germinated seeds after 3-day germination were more complete with higher purity.Using the total genomic DNA of one dried onion seed extracted by CTAB method as the template,SCAR markers were used to identify the Ms loci genotypes of onion hybrids.The PCR amplified bands were clear and bright,and the differences between different nuclear genotypes of onion could be clearly distinguished according to the migration positions of the bands.In summa-ry,this study determined that CTAB method was the best extraction method for the total genomic DNA from dried onion seeds,and the effective DNA obtained from one dried onion seed could satisfy the requirements of hybrid molecular marker identification,and realize the early identification of hybrid purity.
国家科技期刊平台
NSTL
万方数据
维普
