Application of Inter-Simple Sequence Repeat Markers in Genetic Variance Detection of Proso Millet (Panicum miliaceum L.) Germplasm Cultivated in Kazakhstan

This study examined the genetic diversity and relationships among 67 proso millet genotypes, comprising different varieties from eight origin groups. Panicum miliaceum L. is a highly cultivated crop in Kazakhstan. However, there has been no comprehensive study in Kazakhstan to investigate the genetic diversity and phylogenetic relationships of millet using intersimple sequence repeat (ISSR) markers. Genomic DNA from the proso millet genotypes was analyzed using ISSR markers. A total of 16 ISSR primers generated 4268 amplified DNA fragments, ranging in size from 43 to 4792 bp (base pairs). Of these fragments, 901 were monomorphic (29.7%), while 3367 were polymorphic (70.3%), with an average polymorphism of 78.8% per primer. Primers 834, 835 and 840 exhibited the highest polymorphism and were the most effective in distinguishing the genetic diversity of the proso millet accessions. Genetic diversity parameters were evaluated for the eight groups, revealing an average polymorphism information content (PIC) value of 0.124. Principal coordinate analysis (PCoA) and the phylogenetic tree demonstrated a wider genetic diversity among the 67 accessions from the eight groups. Analysis of molecular variance (AMOVA) revealed the presence of extensive genetic variability within groups and minor molecular difference among the studied groups. This research provides novel insights into germplasm classification, evolutionary relationships, genomics, and breeding of proso millet. © 2025 Elsevier B.V., All rights reserved.