The phyllosphere represents the largest biological surface on Earth and serves as an untapped reservoir of functional microbiota. The phyllosphere microbiome has the potential to mitigate plant diseases; however, limited information exists regarding its role in maintaining plant health. In this study, metagenomic sequencing was employed to analyze the microbiomes of the adaxial and abaxial leaf surfaces of healthy (CKWT) and diseased (EWT) maize, with the aim of dissecting the influence of Exserohilum turcicum on phyllosphere microbiome function. E. turcicum altered the alpha and beta diversity of the phyllosphere microbiome, with the Shannon diversity and Chao1 index values significantly reduced in EWT. More beneficial microbes accumulated in the CKWT phyllosphere, whereas pathogenic microbes decreased. E. turcicum may have altered the balance between commensal and pathogenic microorganisms. The species and abundances of microorganisms on the two sets of leaf surfaces were also altered after inoculation with E. turcicum. Further analysis of disease-resistance-related metabolic pathways and abundances of antibiotic-resistance genes revealed that E. turcicum altered the abundance of the functional microbiome and modified the microbiome differences between adaxial and abaxial leaf surfaces. In conclusion, the results reveal that microbial diversity in the maize phyllosphere can influence the microbiome and regulate microbial functions to support plant health. These findings enhance our understanding of how E. turcicum affects the phyllosphere microbiome and provide a theoretical basis for biological control of E. turcicum.

The DIR gene family, which encodes Dirigent proteins, plays a crucial role in plant development and stress responses. However, the functions and mechanisms of this family in maize remained underexplored. This study identified ZmDIR11, a member of the maize Dirigent protein family, and explored its role in drought tolerance. ZmDIR11 shared conserved regions with homologous proteins in wheat, rice, and Arabidopsis. RT-qPCR analysis revealed that ZmDIR11 expression is upregulated in leaves under drought and PEG stress, while subcellular localization confirmed its presence in the endoplasmic reticulum. Virus-induced gene silencing (VIGS) and EMS mutagenesis demonstrated that ZmDIR11 silencing or mutation significantly reduces drought tolerance in maize seedlings, indicating its positive regulatory role. Silencing or mutation of ZmDIR11 led to decreased growth parameters (plant height, root length, fresh weight, dry weight, and chlorophyll content) under drought stress, alongside a reduced antioxidant capacity, as evidenced by increased levels of MDA and ROS and decreased activities of SOD, CAT, and POD. Furthermore, ZmDIR11 mutation impaired the synthesis of ABA and zeatin, downregulating key genes in the ABA (ZmABA1, ZmNCED3, ZmSnRK2.6) and zeatin (ZmIPT4, ZmCKO5, ZmCKO4b) pathways. Drought-responsive genes (ZmRD20, ZmRD22, ZmDREB2A) and lignan biosynthesis genes (Zm4CL, ZmC3H, ZmCAD1) are also downregulated in ZmDIR11 mutants. In conclusion, ZmDIR11 enhances drought tolerance in maize by regulating antioxidant capacity, ABA and zeatin synthesis, and lignan metabolism. This study provides new insights into the role of DIR genes in drought tolerance and offers a potential genetic resource for breeding drought-resistant maize varieties.

Introduction: Maize seedling blight, caused by the phytopathogenic fungus Fusarium verticillioides , is a common and rapidly spreading disease that reduces grain quality and yield. Control of this pathogen is challenging due to its complex infection process and the development of resistance to chemical pesticides. Biological control agents are therefore recognized as a sustainable and environmentally friendly strategy for the maize seedling blight. Results: In this study, we isolated and identified Paenibacillus peoriae 3-B4 from maize leaves, which exhibited a 59.92% inhibitory rate against F. verticillioides in greenhouse co-inoculation conditions. Genome sequencing revealed a 5.91 Mb chromosome containing biosynthetic gene clusters involved in antifungal secondary metabolites, along with genes associated with induced systemic resistance (lSR) and pattern-triggered immunity (PTl). Transcriptomic analysis was performed on maize seedling leaves from a greenhouse experiment, identifying 8,997 differentially expressed maize genes, which were primarily enriched in MAPK signaling and plant–pathogen interactions. 16S rRNA analysis further showed significant shifts in microbial communities, particularly an increase in the abundance of beneficial genera such as Paenibacillus, Delftia , and Corynebacterium . The combined analysis of key differentially expressed genes (DEGs) and bacterial community profiles suggested coordinated shifts in host transcriptional responses and microbial abundances following P peoriae 3-B4 treatment. Discussion: These findings not only emphasize the importance of endophytic bacteria in plant defense, but also provide new insights into the development of sustainable biological control strategies for maize fungal diseases.

Fusarium fujikuroi is the primary causal agent of rice bakanae disease, which can lead to substantial yield losses. Developing a rapid, highly specific, and accurate method for detecting F. fujikuroi is crucial for effective surveillance, prevention, and control of rice bakanae disease. In this study, a novel detection assay, RPA-Cas12a-F, was developed by integrating recombinase polymerase amplification (RPA) and Cas12a for the detection of F. fujikuroi. This assay demonstrated a limit of detection (LOD) of 1 copy/μL of reference plasmid or 0.1 fg/μL of F. fujikuroi genomic DNA (gDNA). Furthermore, to enable on-site detection, the RPA-Cas12a technique was combined with a lateral flow strip (LFS) for visual readout, thereby developing the RPA-Cas12a-LFS assay. The LOD of the RPA-Cas12a-LFS assay was 1000 copies/μL of plasmid or 10 fg/μL of F. fujikuroi gDNA. The RPA-Cas12a-based assays developed in this study enable rapid, highly accurate, sensitive, and specific detection of F. fujikuroi, making them a promising tool for on-site detection without the need for expensive equipment and time-consuming methodologies.

Drought and salt stresses are major abiotic factors limiting maize yield. Lignin, a key cell wall component, plays a crucial role in boosting plant stress resistance. Cinnamyl alcohol dehydrogenase (CAD) is a vital enzyme at the late stage of lignin biosynthesis; however, a systematic study of its functions in abiotic stress responses and its potential for genetic improvement in maize remains lacking. In this study, we conducted the first comprehensive, multi-dimensional analysis of the maize ZmCAD gene family, including gene identification, evolutionary relationships, protein interaction networks, and stress-responsive expression patterns. We identified 9 ZmCAD members that showed significant functional divergence in evolution, structure, and expression patterns. Expression analysis revealed complex, tissue-specific responses of ZmCAD genes to drought and salt stress, with ZmCAD6 strongly induced by drought. Importantly, through haplotype analysis of 157 waxy maize inbred lines, we successfully identified an elite haplotype (H3) of ZmCAD6 that is significantly associated with improved drought tolerance in maize. This study not only clarifies the functional differentiation mechanisms of the ZmCAD gene family but also provides the identified elite ZmCAD6-H3 haplotype as a valuable genetic resource and precise target for molecular breeding aimed at enhancing drought tolerance in maize.

Objectives: To facilitate the innovation and efficient utilization of specialty maize germplasm, this study aimed to systematically evaluate a panel of 222 inbred lines. The objective was to comprehensively characterize phenotypic variation, genetic diversity, and genotype–phenotype associations to screen for representative germplasm resources. Methods: We integrated Best Linear Unbiased Prediction (BLUP) values derived from multi-environment field trials with high-density whole-genome single-nucleotide polymorphism (SNP) data. Population structure and genetic diversity were analyzed, Mantel tests were conducted to assess genotype–phenotype correspondence, and a genome-wide association study (GWAS) was performed to identify significant loci. Results: The population exhibited substantial phenotypic variation, particularly in plant height and tassel traits, with distinct morphological differentiations among specialty types. Genetic diversity analyses revealed varying diversity levels among subpopulations. While Mantel tests indicated a weak overall genotype–phenotype correspondence, specific traits showed significant associations with genetic distance. GWAS successfully identified significant loci associated with plant height and tassel traits. Furthermore, population structure analysis revealed distinct genetic stratification corresponding to specialty types, albeit with a certain degree of admixture. Conclusions: By integrating multi-dimensional phenotypic and genomic profiles, a panel of highly diverse and representative candidate germplasm was identified. These findings provide a crucial theoretical basis for specialty maize breeding and the optimized utilization of germplasm resources.