Maize Purity And Paternity Testing Using InDel Markers

Zea mays L.is one of the most important crops in the world.Compared with other crops such as rice and wheat,it not only has the function of food,but also has the functions of chemical raw materials,energy and feed,thus ensuring the increase of maize production is of great significance to ensure food security and improve people's living standards in our country.With the development of China's seed market,protection of maize variety rights and variety management are becoming more and more important.In the process of seed production,the purity of hybrids is reduced due to lax isolation,untimely emasculation,and incomplete removal of impurities.Therefore,it is urgent to develop and establish a high-throughput and low-cost purity identification method for maize hybrids to keep the healthy development of our country's maize production.Addationally,in the current identification,the submitting party is often unwilling or unable to provide real parental seeds due to variety protection and commercial secrets,so it is also necessary to establish an identification method for obtaining the DNA fingerprint of the parent from the F1 generation.In this study,maize hybrids and their parents were used as research materials,and a new dual-platform purity detection process for hybrids and a parental genotype separation process using endosperm were formulated using InDel molecular markers.The main results of the study are as follows:1.The fingerprint of maize hybrids and their parents were obtained based on 20 pairs of InDel molecular markers developed by the Maize Research Center of Beijing Academy of Agriculture and Forestry.The parental complementation locis were screened out and the purity identification of the artificially created purity samples was carried out on the KASP platform.The results shows that the statistical results of the genotyping were consistent with expectations,proving that maize hybrids purity identification can be successfully carried out based on the InDel combined with the fluorescent capillary platform and the KASP platform.2.Based on the 2:1 dose relationship between the maternal and paternal DNA in the maize endosperm,20 InDel molecular markers were used to perform paternity testing on a fluorescent capillary electrophoresis platform to obtain the triplet fingerprint data of maize hybrids.The peak area of each InDel genotype and the allele amplification ratio of each embryo and endosperm were obtained from the embryo and endosperm DNA of 8maize hybrids,respectively.It was found that there were significant differences in the allele amplification ratio between the embryo and the endosperm,and the fingerprint data of the triplet of the corn hybrid was finally obtained.By comparing with the fingerprints of the parents of the hybrid,the correctness of the method of splitting the genotypes of the parents using the endosperm was verified.