## 3 Results
### Transcriptome Expression Pattern Analysis  
To further elucidate the gene expression profiles in response to pathogen inoculation, we conducted transcriptome sequencing on RNA samples extracted from the leaves of N531_R and N531_S. These samples were collected at multiple time points, immediately before inoculation (0 h) and at 12, 24, 48, and 96 h post-inoculation. A total of 3514 genes were significantly enriched across the five time points, with the majority of expressed genes downregulated in N531_S compared to N531_R (Figure 4a). The expression patterns of the differentially expressed genes (DEGs) in the leaves of N531_R and N531_S across the five sampling stages were visualized using hierarchical clustering (Figure 4b). The expression profiles of the same samples were generally clustered together, such as N531_R samples at 48 and 96 h and N531_S samples at 0, 24, 48, and 96 h showing similar patterns. Notably, the expression profiles of the N531_R samples clustered with the earlier stages of inoculation of the N531_S samples. For instance, the N531_R samples at 12 h clustered with the sample of N531_S at the same point. Further functional categorization of the DEGs was performed according to the Gene Ontology (GO) terms using GOseq tools. The representative categories included defense mechanisms; secondary metabolite biosynthesis, transport, and catabolism; and signal transduction mechanisms. We conducted a hierarchical clustering analysis on differentially expressed gene samples, and the Venn diagram results showed that 15 differentially expressed genes were identified at 12 h, 24 h, 48 h, and 96 h after infection (Figure 4c; Supplementary Materials S2). Additionally, we identified 11 related genes exhibiting similar expression patterns to those of the target candidate genes _Zmays10G000440_ (the highest confidence resistance gene) (Figure 4d; Supplementary Materials S3). Among these, _Zmays01G009610_ was identified as DNAJ heat shock protein 40 (hsp40). After induction, significant differences in expression levels were observed between N531_R and N531_S at 24 and 48 h post-infection [37; 38]. _Zmays01G042730_, which shares similarities with _AtPRL1_, a WD40-containing protein, was significantly induced in N531_S during the first 12 and 24 h following infection [39] (Supplementary Materials S4). This gene may be associated with the enrichment of reactive oxygen species in N531_S.  
Figure 3: The structure of the candidate genes and their expression levels in _RppSLN_. (**a**) The diagram depicts the structure of _RppSLN_ in N531_R, situated between N531 and K22. The top panel reveals that the susceptible line N531 harbors one NBS-LRR gene, noted as _NR5_. The middle panel shows the two candidate genes of _RppSLN_ in the resistant line N531_R, annotated as _Zmays10G000430_ and _Zmays10G000440_. The bottom panel indicates that the K22 contains three NBS-LRR genes known as _R1_, _R2_, and _R3_. (**b**) An expression analysis of _Zmays10G000430_ and _Zmays10G000440_ and an expression analysis of _NR5_. The values presented are the means \(\pm\) standard deviations of the data. For (**b,c**), the different letters in the columns indicate a significant difference (_p_ < 0.05), as determined by the Duncan test.