Drought is considered the most hazardous environmental abiotic stress factors, which adversely affects wheat production (Triticum aestivum L.). Several transcription factor (TF) families, such as WRKY, MYB, and NAC, have an important role in response to drought stress in plants. The investigation of NaC and WRKY genes is considered a new area for research, especially in Egypt. Thus, this study aimed to detect the three genes namely, TaNAC2a, TaWRKY2, and TaWRKY19 by using DNA marker based on encoding regions in different wheat genotypes. In addition to that, to identify the polymorphism in TaNAC2a gene by performing sequence analysis of the central part of exon 2 in seven wheat cultivars. The TaNaC2a, TaWRKY19, and TaWRKY2 genes were detected in most of the tested wheat genotypes. The polymorphic bands detected for TaNAC2a, TaWRKY19, and TaWRKY2 suggest the existence of other alleles at the same locus of these genes. The central part of exon2 was sequenced (260 bp) showing the conserved sequence in all cultivars, except the positions from 813 to 863 were high polymorphic. Detected SNPs related to drought tolerance were not found. The amino acids sequence encoded from the target DNA region was highly conserved from position 282 to 319 in the N-terminal domain in all genotypes except a few positions. Detection of these important genes is considered a critical start point for further study especially gene expression of these genes.

Ensuring sustainable wheat production is crucial for food security, particularly in regions facing plant diseases and environmental challenges. In the Middle East, wheat powdery mildew, caused by Blumeria graminis f. sp. tritici, significantly threatens yields and quality, making it essential to comprehend local pathotype virulence and resistance genes for effective disease management. Therefore, 252 infected leaves were sampled from cultivated bread wheat genotypes in Egypt Delta region during 2024–2025 growing seasons. Forty-one powdery mildew monogenic lines (Pm) and twelve new bread wheat genotypes were evaluated at seedling and adult stages. Pathotype analysis identified six pathotypes in 2024 (IOEE most common, 22.91%) and four in 2025 (POKE dominant, 34.25%). Virulence diversity declined significantly: Shannon diversity decreased from 5.90 to 3.73, and Simpson diversity decreased from 5.81 to 3.58, indicating reduced pathotype complexity. No virulence was detected against Pm11, Pm13, Pm24, Pm25, Pm29, Pm32, Pm34, Pm35, Pm36, Pm37, Pm43, NCA9, and MIAB10 genes. PERMANOVA analysis showed no significant compositional difference between seasons (R² = 0.112, p = 0.481), with low genetic differentiation (mean character difference = 0.1894; Roger’s distance = 0.1914). Sakha96 wheat genotype displayed complete resistance at both growth stages, while Misr4, Misr5, Sakha95, Sids15, and Gemmeiza13 showed adult-stage resistance only in both seasons. Molecular analysis revealed that Pm24, Pm35 and Pm37 were absent in all genotypes. Pedigree analysis demonstrated Pm35 and Pm37 share common ancestral origins across Egyptian wheat germplasm. So, these resistant genotypes and broadly effective resistance genes provide valuable resources for breeding durable powdery mildew resistance in Egyptian wheat.