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.

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.

Drought stress critically endangers the growth and development of crops. Glutathione S-transferase plays a vital role in response to abiotic stress. However, there are few studies on the role of glutathione S-transferase in maize drought stress. In this study, the significantly downregulated expression of ZmGST26 in roots under drought stress was analyzed by qRT-PCR. Promoter analyses showed that there were several cis-acting elements related to drought stress and that were involved in oxidative response in the promoter region of ZmGST26. Subcellular localization results showed that ZmGST26 was localized in the nucleus. The transgenic lines of the Arabidopsis over-expressing ZmGST26 were more sensitive to drought stress and ABA in seed germination and inhibited ABA-mediated stomatal closure. Under drought stress, phenotypic analyses showed that the germination rate, root length and survival rate of ZmGST26 overexpressing lines were significantly lower than those of wild-type lines. The determination of physiological and biochemical indexes showed that the water loss rate, malondialdehyde, O₂⁻ and H₂O₂ of the overexpression lines significantly increased compared with wild-type Arabidopsis, but the antioxidant enzyme activities (CAT, SOD and POD), and proline and chlorophyll contents were significantly reduced. Subsequently, the qRT-PCR analysis of drought stress-related gene expression showed that, under drought stress conditions, the expression levels of DREB2A, RD29A, RD29B and PP2CA genes in ZmGST26 overexpression lines were significantly lower than those in wild-type Arabidopsis. In summary, ZmGST26 reduced the drought resistance of plants by aggravating the accumulation of reactive oxygen species in Arabidopsis.

Drought stress is one of the major adversity stresses affecting soybean (Glycine max [L.] Merr.) yield. Late embryogenesis abundant protein (LEA protein) is a large family of proteins widely distributed in various types of organisms, and this class of proteins plays an important role in protecting proteins, membrane lipids, and lipids inside the cell. The soybean GmPM35 gene is a member of the LEA_6 subfamily. The expression of the GmPM35 gene was significantly increased after drought stress in soybeans. A subcellular localization assay confirmed that the gene acts on the cell membrane. Against wild-type Arabidopsis thaliana, we found that Arabidopsis lines overexpressing the GmPM35 gene were significantly more drought-tolerant at germination and seedling stages under drought stress. To further investigate the drought tolerance function of this gene in soybeans, nine overexpression lines of the T₃ generation soybean GmPM35 gene and two editing lines of the T₃ generation soybean GmPM35 gene were obtained by Agrobacterium-mediated method using a wild-type soybean strain (JN28) as a receptor. Germination rate, root length, chlorophyll (CHL) content, Proline (Pro) content, malondialdehyde (MDA) content, superoxide anion (O₂^•−) content, hydrogen peroxide (H₂O₂) content, (NBT, DAB) staining, and activities of antioxidant enzymes (CAT, SOD, POD), and photosynthetic physiological indexes of the three different types of strains were measured and analyzed before and after drought stress. Combined with the results of rehydration experiments and physiological and biochemical indices, we found that overexpression of the GmPM35 gene protected the activities of antioxidant enzymes under drought stress. The activities of superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) were increased by an average of 34.28%, 26.12%, and 30.01%, respectively, in soybean plants overexpressing the GmPM35 gene compared with wild-type soybeans. Under drought stress conditions, soybean plants overexpressing the GmPM35 gene showed an average increase of 76.81% in photosynthesis rate (Pn), 39.8% in transpiration rate (Tr), 126% in stomatal conductance (Gs), 47.71% in intercellular CO₂ concentration (Ci), and 26.44% in instantaneous water use efficiency (WUEi). The improvement of these indexes helped to reduce the accumulation of reactive oxygen species (ROS) in the plants. In addition, we found that under drought stress, the MDA content was reduced by an average of 18.8%, and the Pro content was increased by an average of 60.14% in soybean plants overexpressing the GmPM35 gene, and the changes in these indexes indicated that the plants had stronger antioxidant and osmoregulatory capacities in response to drought stress. In summary, this experiment demonstrated that the GmPM35 gene plays an important role in soybean tolerance to drought stress, and by overexpressing the GmPM35 gene, soybean plants can better tolerate drought stress and maintain normal physiological functions.

(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.