Genetic Diversity Assessment of Citrus Varieties Using Morphological, Biochemical, and Simple Sequence Repeats Marker Approaches

Citrus is a genetically diverse genus widely cultivated across tropical and subtropical regions, yet its varieties often exhibit overlapping morphological traits that complicate accurate identification. In this study, the genetic diversity of nine Citrus varieties was assessed using 36 Simple Sequence Repeats (SSR) markers in combination with key morphological and biochemical traits. The CTAB method was employed to extract genomic DNA, followed by polymerase chain reaction (PCR) using a specific set of primers. Gel documentation was performed to score the bands obtained. This study aimed to utilize 36 SSR markers to characterize the genetic diversity of nine Citrus varieties collected from the Citrus Research Institute (CRI), Sargodha. Polymorphic information content (PIC) values ranged from 0.20 to 0.50, averaging 0.27, indicating moderate genetic variability among the varieties. Principal component analysis explained 68.7% of the total variation across the first two components, and cluster analysis grouped the varieties into two major clusters, demonstrating clear genetic differentiation. The integration of molecular and morphological data revealed distinct similarity groups, providing valuable insights for selecting superior parental lines. These findings support the development of improved Citrus varieties and the establishment of DNA‐certified true‐to‐type nursery plants for enhanced orchard productivity.