Wheat is a globally important food crop, supporting the food security of approximately 30% of the world’s population. This study aims to systematically identify the SPFH family in common wheat (Triticum aestivum L.), comprehensively analyze its sequence characteristics, protein structure, and response patterns to biotic stress, and reveal its potential biological functions in wheat stress resistance. By integrating genomic data from Arabidopsis thaliana, rice (Oryza sativa L.), and maize (Zea mays) for comparative analysis, this study identified 60 SPFH family members at the whole-genome level in wheat and conducted a comprehensive analysis of their physicochemical properties, gene structure, conserved motifs, cis-regulatory elements, phylogeny, synteny, and ability to form heterocomplexes. The research reveals that SPFH family proteins generally exhibit acidic and hydrophilic characteristics; gene structure analysis shows that the length and number of introns are similar within subfamilies, while significant heterogeneity in sequence length exists between subfamilies. Promoter analysis further reveals that this family is enriched with cis-regulatory elements associated with growth, development, and responses to multiple stresses. Evolutionary analysis based on phylogeny and structural modeling yielded highly consistent results, revealing structural conservation within subfamilies and a convergent evolutionary pattern across the family.Intraspecific and cross-species synteny analyses indicate that the family underwent translocation and gene expansion during polyploidization, with a general 1:3 orthologous ratio between monocots and wheat. Nucleotide diversity and heteropolymerization analysis highlight the extreme conservation of the TaHIR, TaSLP, and TaPHB subfamilies and predict the functional potential of most members to form stable heterocomplexes. Integrated analysis of transcriptome and qRT-PCR data reveals complex expression dynamics of SPFH family members under wheat stripe rust (Puccinia striiformis f. sp. tritici) stress：while transcript levels generally decreased at the macro-transcriptional level, key members were significantly upregulated in the susceptible material Avocet S. In summary, the SPFH family is highly conserved in evolution and sensitive to biotic stress responses. This study provides important candidate targets for elucidating molecular mechanisms of disease resistance in wheat and offers theoretical support for improving stress resistance in crops.