Transcription factors (TFs) are central to transcription in eukaryotes, allowing proper gene regulation across different developmental stages. However, research on TFs is limited in the oomycete Phytophthora infestans, a notorious plant pathogen causing late blight diseases. This study illuminates the DNA-binding preferences of TFs in P. infestans and helps identify genes regulated by a P. infestans MADS-box TF (PiMADS) that regulates sporulation. Genome-wide identification of TFs in P. infestans revealed 197 genes representing 22 TF families. The family expansions appeared to have resulted from ancient events of unequal crossing-over. DNA binding specificities of 73 TFs were defined using protein binding oligonucleotide microarrays (PBM). Consistent with the substantial evolutionary divergence of oomycetes from traditional model organisms, only a subset of the DNA-binding preferences resembled those of human or plant orthologs. Paralogs with similar binding preferences often had different expression patterns signaling distinct functions. A reporter gene study confirmed a Heat Shock Factor TF as a general activator, aligning with the unbiased presences of its motif within promoters of genes with diverse expression patterns. To further understand TFs, ChIP coupled with sequencing was used to identify genome-wide binding sites of PiMADS, a sporulation regulator. A total of 299 binding sites that occurred in the promoter regions were assigned to 316 genes of which 196 were sporulation-induced. Although all 196 genes are sporulation-induced, they exhibited several patterns during sporulation time-courses. The most enriched motifs in ChIP-binding sites resembled the motif from PBM data and the consensus motif for MADS-box proteins of plants and humans. Also, a reporter gene study confirmed that PiMADS bound this motif and was autoregulated. Five out of seven selected PiMADS targets were successfully knocked out using CRISPR-Cas12a. Knock-out strains of a Myb TF no longer formed cleavage planes and released zoospores confirming the TF role in early zoosporogenesis suggestive of PiMADS regulation beyond sporulation. The results from this study offer insights into the regulatory network of TFs in P. infestans in terms of TF binding specificities and regulation of TF target genes.