Genetic Diversity Study of Steudneri Clover (Trifolium steudneri) Accessions of Ethiopia Using Inter Simple Sequence Repeat (ISSR) Markers

Abstract

Inter simple sequence repeat (ISSR) markers were employed to study the genetic diversity of 48 accessions of T. steudneri, representing two populations from Ethiopia. DNA was extracted from a bulk of fresh young leaves from three plants per accession using a modified CTAB method. Four selected di-nucleotide ISSR primer combinations generated a total of 65 bands (200-1000bp), of which 63 (96.92%) were polymorphic. The Nei’s gene diversity (h) and Shannon’s index (I) were high at species level (h = 0.26, I = 0.39). But, the genetic diversity was relatively low at population level, and average within-population diversity was h = 0.22, I = 0.33. Overall, Gojam population showed higher genetic diversity than Shewa. Clustering using UPGMA and PCo based on jaccards coefficient of similarity grouped the accessions according to the geographical origin where they were sampled. The study clearly indicates the presence of variable genotypes with their unique identity that deserve conservation attention.

Keywords: DNA extraction, Molecular markers, Polymerase chain reaction (PCR)