Foxtail millet (Setaria italica), a vital cereal crop in China, serves as both a staple food and forage source but is threatened by rust disease caused by Uromyces setariae-italicae (Usi), leading to severe yield and quality losses. The NAM, ATAF1/2, and CUC2 (NAC) transcription factor family represents one of the largest plant-specific regulatory gene families, playing pivotal roles in development and stress responses. However, the functional relevance of NAC genes in foxtail millet’s defense against this pathogen remains unexplored. Here, we systematically analyzed 33 SiNAC genes from the foxtail millet genome. Phylogenetic analysis classified these genes into 11 subgroups, while chromosomal mapping localized them to nine chromosomes unevenly. Promoter analysis identified stress- and plant hormone-related cis-elements, suggesting functional diversity. Expression profiling analysis showed that most SiNAC genes exhibit tissue-specific expression patterns. Quantitative real-time PCR (qRT-PCR) results indicated that 30 genes responded to Usi infection, with 17 showing a strong association with rust resistance. Three resistance-associated genes demonstrated transactivation activity and nuclear localization, indicating their regulatory function in defense responses. This study provides both mechanistic insights into SiNAC-mediated rust resistance and potential targets for molecular breeding in foxtail millet.

Foxtail millet blast, caused by Pyricularia oryzae, is a widespread airborne disease that severely threatens the sustainable production and industrial development of foxtail millet. Developing eco-friendly and non-toxic management strategies has been widely acknowledged as a core prerequisite for establishing sustainable disease control systems. In this study, a bacterial strain NR2065B9 with prominent in vitro antagonistic activity against P. oryzae was isolated from healthy foxtail millet rhizosphere soil. Taxonomic identification via morphological characterization and 16S rRNA gene sequence homology analysis confirmed that this strain belonged to the species Pseudomonas silesiensis. Dual culture assays revealed that strain NR2065B9 exerted a significant inhibitory effect on the mycelial growth of P. oryzae. Detached-leaf inoculation assays demonstrated that strain NR2065B9 could remarkably restrict the expansion of blast lesions on foxtail millet leaf tissues. Field trials confirmed that NR2065B9 application could reduce foxtail millet blast severity compared with the controls. Collectively, these findings indicated that the antagonistic strain NR2065B9 possessed great application potential in the biocontrol of foxtail millet blast, and could be further exploited as a promising microbial resource for the development of biocontrol agents against this disease.