Striga hermonthica, a parasitic weed, poses a significant threat to sorghum production in sub-Saharan Africa, potentially causing yield losses of up to 100%. Identifying sources of Striga resistance trait within the diverse wild sorghum accessions is imperative to developing resistant sorghum cultivars. This study analyzed the genetic variation of 255 Sudanese wild sorghum accessions using simple sequence repeat (SSR) markers associated with post-attachment Striga resistance mechanisms: nine SSR markers linked to mechanical barrier resistance and two to hypersensitive resistance. We found moderate polymorphism in the Striga resistance loci among the Sudanese wild sorghum accessions, with polymorphic information contents (PIC) ranging from 0.03 to 1.92 for hypersensitivity and 0.37 for mechanical barrier resistance mechanism markers. Overall, 18 alleles were detected within the loci for mechanical barrier markers and four for hypersensitive markers. A population structure and cluster analysis revealed that several accessions were closely linked to the resistant checks N13 (mechanical barrier) and Framida (hypersensitivity response). Analysis of molecular variance (AMOVA) showed substantial polymorphism within the population (99% for mechanical barriers and 94% for hypersensitivity), thus indicating that these wild sorghum accessions harbor ready-to-use genes for improving Striga resistance in sorghum. Our findings highlight the merits of Sudanese wild sorghum germplasm for post-attachment Striga resistance mechanisms, indicating their possible use in sorghum breeding efforts to develop Striga resistant cultivars.