Fluorescent SSR-Based DNA Fingerprinting and Molecular Identity Card Development for 69 Mandarin Accessions

To establish standardized DNA fingerprinting and molecular identification systems for citrus, we analyzed 69 mandarin accessions via fluorescent SSR capillary electrophoresis to construct DNA molecular fingerprints and unique molecular identity cards. Eighteen highly polymorphic SSR primer pairs were screened, yielding 239 genotype calls and 147 alleles. The number of amplified alleles per primer pair ranged from 4 to 18, with polymorphic information content (PIC) values varying from 0.411 to 0.650. Ten core primer pairs were further selected, achieving a discrimination rate of 65.2% (45 out of 69 accessions distinguished). Utilizing these fluorescent SSR markers, we established DNA molecular fingerprints and unique molecular identity cards for all 69 accessions. Among them, 45 accessions possessed unique fingerprints, whereas the remaining 24 indistinguishable accessions were clustered into six groups. Each cluster contained both wild (4 accessions total) and cultivated (20 accessions total) resources with high genetic similarity, which merits further investigation. This study lays a theoretical basis for the authentication, conservation, and genetic relationship analysis of mandarin germplasm resources, and provides a practical tool for standardizing mandarin variety identification.