Maize is an essential staple food, and its genetic diversity plays a central role in breeding programs aimed at developing climate-adapted cultivars. Constructing a representative core germplasm set is necessary for the efficient conservation and utilization of maize genetic resources. In this study, we analyzed 588 cultivated maize accessions using agronomic traits such as plant morphology and yield traits such as ear characteristics and single-nucleotide polymorphisms (SNPs) to assess molecular diversity and population structure and to construct a core collection. Nineteen phenotypic traits were evaluated, revealing high genetic diversity and significant correlations among most quantitative traits. The optimal sampling strategy was identified as “Mahalanobis distance + 20% + deviation sampling + flexible method.” Whole-genome genotyping was conducted using the Maize6H-60K liquid phase chip. Population structure analysis, principal component analysis, and cluster analysis divided the 588 accessions into six subgroups. A core collection of 172 accessions was selected based on both phenotypic and genotypic data. These were further evaluated for salt–alkali tolerance during germination, and cluster analysis classified them into five groups. Sixty-five accessions demonstrated salt–alkali tolerance, including 18 with high resistance. This core collection serves as a valuable foundation for germplasm conservation and utilization strategies.

MYB transcription factors constitute a diverse and functionally versatile family, playing central roles in regulating plant responses to a range of abiotic stressors. Based on previous research, we identified and characterized a maize MYB transcription factor gene, ZmMYBR24, which is involved in responses to salt, alkali, and low-temperature stress. This study aimed to investigate the function and mechanism of ZmMYBR24 in response to salt, alkali, and low-temperature stresses. We hypothesized that ZmMYBR24 regulates biosynthetic pathways to influence maize resistance to multiple abiotic stresses. The results indicate that ZmMYBR24 expression was markedly upregulated (p < 0.01) and the fold-change in gene expression ranged from 1.54 to 25.69 when plants were exposed to these combined stresses. Phenotypically, the zmmybr24 mutant line exhibited more pronounced inhibition of seedling and root growth under stress compared to the wild-type B73 line. Based on a correlation expression pattern analysis and mutant line evaluation, ZmMYBR24 was confirmed to be a positive regulatory transcription factor for multiple types of abiotic stress resistance. An RNA-seq analysis of both lines revealed differentially expressed genes (DEGs), with gene ontology (GO) and KEGG enrichment analyses indicating that ZmMYBR24 may mediate stress responses by modulating the expression of genes involved in flavonoid biosynthesis. Notable differences were observed in the expression of pathway-associated genes between the mutant and wild-type plants. A haplotype analysis across 80 inbred maize lines revealed 16 ZmMYBR24 coding region haplotypes—comprising 25 SNPs and 17 InDels—with HAP12 emerging as a superior haplotype. These results demonstrate that ZmMYBR24 enhances maize yields by regulating the flavonoid biosynthesis pathway in response to adverse climatic conditions including salt, alkaline conditions, and low temperatures. Collectively, these findings offer novel insights into the molecular mechanisms underlying maize adaptation to combined abiotic stresses and lay the groundwork for breeding programs targeting multi-stress resistance.

GLABROUS1 enhancer-binding protein (GeBP) family members are plant-specific transcription factors involved in growth, development, and environmental responses. However, the characteristics and stress-responsive patterns of this family in maize (Zea mays L.), particularly during seed aging and saline–alkali stress, remain insufficiently understood. We identified 19 ZmGeBP genes and classified their encoded proteins into four phylogenetic groups together with homologs from sorghum and rice. Duplication and synteny analyses showed that segmental duplication was the predominant duplication pattern detected in maize and that collinear relationships were retained among the three grasses. The ZmGeBP promoters contained cis-acting elements associated with development, light, phytohormone responses, and stress. RT-qPCR analysis of seven selected genes revealed temporal and genotype-dependent expression patterns during artificial seed aging and saline–alkali stress. Subcellular localization confirmed nuclear targeting of ZmGeBP8, and dual-luciferase reporter assays showed that ZmOCL13 reduced ZmGeBP8 promoter activity, suggesting a potential repressive effect. Overall, these findings elucidate the molecular features of maize ZmGeBP genes and establish a framework for validating their roles in stress resilience.