Triticum aestivum L. is an annual grass of family Poaceae. The present study was concerned with the discrimination and Identification of some Northern African T. aestivum L. cultivars. Fourteen T. aestivum L. cultivars, representing seven Northern African countries, were presented in this study. High-resolution conventional SEM imaging was performed to survey on and to examine the grain surface sculpture (dorsal and ventral views) of the studied cultivars. SEM analysis revealed six grain surface sculpture for the ventral surface and four for the dorsal side. In addition, SCoT polymorphism analysis was performed using 14 primers to access and characterize the genetic diversity between T. aestivum L. cultivars under study. The relationship resulted from applied multidisciplinary analyses viz. (UPGMA, structural analysis, and principal coordinate analysis) via SEM and SCoT analyses were in complement and efficient in assessing the characterization of taxonomic and genetic diversity of studied T. aestivum L. cultivars. In conclusion, it is most likely speculated that breeding lines from T. aestivum cultivars range of North Africa and their rational comparison may emerge novel insights and give better understanding of the domestication of T. aestivum genetic diversity. Also, some cultivars could be used as important genetic resources for genetic improvement of T. aestivum in future breeding program.