Drought is one of the most serious environmental stresses affecting crop production. NAC transcription factors play a crucial role in responding to various abiotic stresses in plants. Here, we identified a maize NAC transcription factor, ZmSNAC06, between drought-tolerant and drought-sensitive inbred lines through RNA-seq analysis and characterized its function in Arabidopsis. ZmSNAC06 had five transcripts, of which ZmSNAC06-T02 had a typical NAC domain, while ZmSNAC06-P02 was localized in the nucleus of maize protoplasts and had transactivation activity in yeasts. The expression of ZmSNAC06 in maize was induced by drought. The overexpression of ZmSNAC06-T02 in Arabidopsis resulted in hypersensitivity to abscisic acid (ABA) at the germination stage, and overexpression lines exhibited higher survival rates and higher antioxidant enzyme activities compared with the wild-type under drought stress. These results suggest that ZmSNAC06 acts as a positive regulator in drought tolerance and may be used to improve drought tolerance in crops.

Drought stress is seriously affecting the growth and production of crops, especially when agricultural irrigation still remains quantitatively restricted in some arid and semi-arid areas. The identification of drought-tolerant genes is important for improving the adaptability of maize under stress. Here, we found that a new member of the actin-depolymerizing factor (ADF) family; the ZmADF5 gene was tightly linked with a consensus drought-tolerant quantitative trait locus, and the significantly associated signals were detected through genome wide association analysis. ZmADF5 expression could be induced by osmotic stress and the application of exogenous abscisic acid. Its overexpression in Arabidopsis and maize helped plants to keep a higher survival rate after water-deficit stress, which reduced the stomatal aperture and the water-loss rate, as well as improved clearance of reactive oxygen species. Moreover, seventeen differentially expressed genes were identified as regulated by both drought stress and ZmADF5, four of which were involved in the ABA-dependent drought stress response. ZmADF5-overexpressing plants were also identified as sensitive to ABA during the seed germination and seedling stages. These results suggested that ZmADF5 played an important role in the response to drought stress.

Genomic prediction enables rapid selection of maize varieties with low kernel water content (KWC), facilitating the development of mechanized maize harvesting and reducing costs. This study evaluated and characterized the KWC and grain yield (GY) of hybrid maize in northern China and used genomic prediction to identify superior hybrid combinations with low kernel water content at maturity (MKWC) and high GY adapted to northern China. A total of 285 hybrids obtained from single crosses of 34 inbred lines from Stiff Stalk and Non-Stiff Stalk heterotic groups were used for genomic prediction of KWC and GY. We tested 20 different statistical prediction models considering additive effects and evaluating the impact of dominance and epistasis on prediction accuracy. Employing 10-fold cross-validation, it showed that the average prediction accuracy ranged drastically from 0.386 to 0.874 across traits and models. Eight linear statistical methods displayed a very similar prediction accuracy for each trait. The average prediction accuracy of machine learning methods was lower than that of linear statistical methods for KWC-related traits, but the random forest model had a high prediction accuracy of 0.510 for GY. When genetic effects were incorporated into the prediction model, the prediction accuracy for each trait was improved. Overall, the model with dominant and epistatic effects (G:AD(AA)) performed best. For the same number of markers, predictions using trait-specific markers resulted in higher prediction accuracy than randomly selected markers. When the number of trait-specific SNPs was set to 100, the prediction accuracy of GY increased by 33.27%, from 0.406 to 0.541. Out of all the 561 potential hybrids, the TOP 30 hybrids selected by genomic prediction would lead to a 1.44% decrease in MKWC compared with Xianyu335, a hybrid with a fast kernel water dry-down, and these hybrids also had higher GY simultaneously. Our results confirm the value of genomic prediction for hybrid breeding low MKWC suitable for maize mechanized harvesting in northern China. In conclusion, this study highlights the potential of genomic prediction to optimize maize hybrid breeding, enhancing efficiency and providing insights into genotype-accuracy relationships. The findings offer new strategies for hybrid design and advancing mechanized harvesting in northern China.

The highly conserved actin depolymerizing factor (ADF) plays an important role in plant growth, development and responses to biotic and abiotic stresses. A total of 72 ADF genes in Arabidopsis, wheat, rice and sorghum can be divided into four groups. The multicollinearity analysis revealed that the maize ADF gene family exhibited more collinearity events with closely related gramineous plants. Fifteen ADF genes in maize were screened from the latest database, and bioinformatics analysis showed that these ADF genes were distributed across seven chromosomes in maize. The gene structure of the ADF gene family in maize exhibits significant conservation and cluster consistency. The promoter region contains rich regulatory elements that are involved in various regulations related to growth, development and adverse stresses. The drought-tolerant ZmADF5 gene in maize was further studied, and it was found that the allelic variations in ZmADF5 were mainly concentrated in its promoter region. A superior haplotype, with drought tolerance, was identified by candidate-gene association analysis of 115 inbred lines. By comparing the phenotypes of anthesis silking interval, grain yield and ear height, it was found that Hap2 performed better than Hap1 under drought stress. This study provides a theoretical reference for understanding the function of the ADF gene family and proposes further investigation into the role of ZmADF5 in abiotic-stress tolerance.