Maize (Zea mays L.) originates from the subtropical region and is a warm-loving crop affected by low-temperature stress. Dehydrin (DHN) protein, a member of the Group 2 LEA (late embryogenesis abundant proteins) family, plays an important role in plant abiotic stress. In this study, five maize DHN genes were screened based on the previous transcriptome sequencing data in our laboratory, and we performed sequence analysis and promoter analysis on these five DHN genes. The results showed that the promoter region has many cis-acting elements related to cold stress. The significantly upregulated ZmDHN15 gene has been further screened by expression pattern analysis. The subcellular localization results show that ZmDHN15 fusion protein is localized in the cytoplasm. To verify the role of ZmDHN15 in cold stress, we overexpressed ZmDHN15 in yeast and Arabidopsis. We found that the expression of ZmDHN15 can significantly improve the cold resistance of yeast. Under cold stress, ZmDHN15-overexpressing Arabidopsis showed lower MDA content, lower relative electrolyte leakage, and less ROS (reactive oxygen species) when compared to wild-type plants, as well as higher seed germination rate, seedling survival rate, and chlorophyll content. Furthermore, analysis of the expression patterns of ROS-associated marker genes and cold-response-related genes indicated that ZmDHN15 genes play an important role in the expression of these genes. In conclusion, the overexpression of the ZmDHN15 gene can effectively improve the tolerance to cold stress in yeast and Arabidopsis. This study is important for maize germplasm innovation and the genetic improvement of crops.

Osmotic stress caused by drought and high salinity is the key factor limiting plant growth. However, its underlying molecular regulatory mechanism remains unclear. In this study, we found the stress-related gene Zm00001d019704 (ZmSRG7) based on transcriptome sequencing results previously obtained in the laboratory and determined its biological function in maize. We found that ZmSRG7 was significantly expressed in both roots and leaves under 10% PEG6000 or 150 mM NaCl. Subcellular localization showed that the gene was localized in the nucleus. The germination rate and root length of the ZmSRG7 overexpressing lines were significantly increased under drought or salt stress compared with the control. However, after drought stress, the survival rate and relative water content of maize were increased, while the water loss rate was slowed down. Under salt stress, the Na⁺ concentration and Na⁺: K⁺ ratio of maize was increased. In addition, the contents of antioxidant enzymes and proline in maize under drought or salt stress were higher than those in the control, while the contents of MDA, H₂O₂ and O₂⁻ were lower than those in the control. The results showed that the ZmSRG7 gene played its biological function by regulating the ROS signaling pathway. An interaction between ZmSRG7 and the Zmdhn1 protein was found using a yeast two-hybrid experiment. These results suggest that the ZmSRG7 gene can improve maize tolerance to drought or salt by regulating hydrogen peroxide homeostasis.

The papain-like cysteine proteases (PLCPs) is a subfamily of cysteine proteases that plays an important role in leaf senescence, and some of its members are involved in the regulation of plant growth and development under stress. In this study, we cloned a new gene, ZmSAG39, from maize. Expression profile analysis showed that ZmSAG39 was induced by darkness and drought treatments. In addition, the ZmSAG39 overexpression in maize accelerated the senescence of maize leaves under darkness and drought treatments. However, the knockout of ZmSAG39 in maize enhanced the resistance of maize to darkness and drought stresses and reduced the degree of senescence of maize leaves. Under drought stress, compared with WT plants, the knockout lines had a higher seed germination rate, seedling survival rate and chlorophyll content, and lower reactive oxygen species (ROS) level and malondialdehyde (MDA) content. In addition, quantitative real-time PCR (qRT-PCR) analysis showed that ZmSAG39 negatively regulated some stress-related genes but positively regulated senescence-related genes under darkness and drought stress conditions. To summarize, these results indicate that ZmSAG39 is a senescence-related gene and plays a negative role in response to darkness and drought stresses. This study laid a theoretical foundation for the innovation of maize germplasm resources with high quality, high yield and strong stress resistance.

As a staple food crop, maize is widely cultivated worldwide. Sex differentiation and kernel development are regulated by auxin, but the mechanism regulating its synthesis remains unclear. This study explored the influence of the growth stage of maize on the secondary metabolite accumulation and gene expression associated with auxin synthesis. Transcriptomics and metabonomics were used to investigate the changes in secondary metabolite accumulation and gene expression in maize leaves at the jointing, tasseling, and pollen-release stages of plant growth. In total, 1221 differentially accumulated metabolites (DAMs) and 4843 differentially expressed genes (DEGs) were screened. KEGG pathway enrichment analyses of the DEGs and DAMs revealed that plant hormone signal transduction, tryptophan metabolism, and phenylpropanoid biosynthesis were highly enriched. We summarized the key genes and regulatory effects of the tryptophan-dependent auxin biosynthesis pathways, giving new insights into this type of biosynthesis. Potential MSTRG.11063 and MSTRG.35270 and MSTRG.21978 genes in auxin synthesis pathways were obtained. A weighted gene co-expression network analysis identified five candidate genes, namely TSB (Zm00001d046676 and Zm00001d049610), IGS (Zm00001d020008), AUX2 (Zm00001d006283), TAR (Zm00001d039691), and YUC (Zm00001d025005 and Zm00001d008255), which were important in the biosynthesis of both tryptophan and auxin. This study provides new insights for understanding the regulatory mechanism of auxin synthesis in maize.

Flowering is an essential process in plant development, and there are six major flowering pathways: the photoperiodic pathway, gibberellin pathway, vernalization pathway, age pathway, autonomous pathway, and temperature pathway. In this study, we screened the transcriptome sequencing of early flowering mutants from the laboratory for the significantly differentially expressed ZmGRAS46, which belongs to the DELLA subfamily of the GRAS family. DELLA is involved in the gibberellin pathway to regulate plant flowering. However, it is not clear whether ZmGRAS46 is involved in the gibberellin pathway which regulates plant flowering; therefore, in this experiment, we investigated the regulatory role of this gene in Arabidopsis flowering by overexpressing ZmGRAS46. It was found that overexpression of ZmGRAS46 in Arabidopsis promotes the formation of rosette leaves and flower buds and delays flowering time in Arabidopsis, and experiments have shown that ZmGRAS46 represses the expression of FLOWERING LOCUS T (FT), SUPPRESSOR OF CONSTANS1 (SOC1), CONSTANS (CO), and LEAFY (LFY). Our results indicated the possibility that ZmGRAS46 represses flowering through the CO-FT-SOC1-mediated photoperiodic flowering pathway. The delayed flowering phenotype of overexpressing ZmGRAS46 Arabidopsis could be rescued by applying GA3. The experimental results indicate that ZmGRAS46 depends on the GA3 pathway to regulate flowering in Arabidopsis.

(1) Corn is the most widely planted food crop, feed crop, and economic crop in the world, and plays an important role in agricultural production and national economy development. The copine gene, also known as the BONZAI gene, encodes a Ca²⁺-dependent phospholipid membrane binding protein that is widely present in eukaryotes. It has been found that the copine protein is a negative regulator of disease resistance regulation and plays a key role in plants’ disease resistance response. In this study, the Agrobacterium-tumefacien-mediated method was used to successfully obtain T2 generation ZmBON3-gene-overexpressing plants and gene-edited plants. Related phenotypes and molecular identification showed that the disease resistance of overexpression plants was significantly reduced, and the disease resistance of gene-edited plants was significantly increased, which verified that the ZmBON3 gene was a negative regulatory gene. By detecting the physiological indexes related to defense, it was found that the content of H₂O₂ and the enzyme active water of CAT, POD, SOD, and PAL in ZmBON3-gene-edited plants was higher than those in the control plants and ZmBON3-gene-overexpressing plants, and the content of H₂O₂ and CAT, POD, and SOD in ZmBON3-gene-overexpressing plants was significantly higher than that in the control plants and ZmBON3-gene-overexpressing plants. The enzyme activity of PAL was the lowest. By detecting the expression of key genes of defense-related signaling pathways, it was found that ZmBON3 may be involved in the related defense processes mediated by the R gene, SA pathway, JA pathway, and ABA pathway. In addition, ZmBON3-geneedited plants showed obvious dwarf phenomenon at the seedling stage, but this did not affect the ear length, axis diameter, ear row number, and grain color.