Development of DNA Molecular Identification Card of Forage Oat Varieties Based on SSR
In order to accurately identify forage oat varieties and solve the problems of variety confusion in production,15 pairs of SSR primers were used to amplify the genomic DNA of 19 forage oat varieties,on the basis of genetic diversity analysis,core primer combinations were screened to construct the molecu-lar fingerprints of the tested varieties and developed their molecular identity cards.The results showed that a total of 70 alleles were detected with 15 pairs of SSR primers,with 2-8 alleles detected at each locus,and the number of effective alleles ranged from 1.6437 to 5.4783,with an average of 2.5741;the observed het-erozygosity(Ho)ranged from 0.3939 to 0.8931,which was higher than that of expected heterozygosity;and the polymorphism information content(PIC)value of primer M08 was the highest(0.8391),and that of primer M07 was the lowest(0.4736).After excluding primers with PIC lower than 0.7,the distinguish-ing rate of the remaining primers to the test varieties was 10.53%to 26.32%.Among the two pairs of primer,M08-M03 and GM8-M03 had the highest distinguishing rate(73.68%),but it was still insuffi-cient to distinguish all varieties.The distinguishing rate of M08-P15-M03 in 3 pairs of primers was 100%,which could completely distinguish 19 tested oat varieties.By using core primers M08-P15-M03,the amplified fragments of each variety were digitally encoded,and each band type code was connected succes-sively according to the fixed sequence of primers,and the molecular identification codes of each variety were successfully constructed and converted into barcodes and two-dimensional codes that could be recog-nized by machines,realizing rapid identification of varieties.
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