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. 2025 Dec;16(1):28-45.
doi: 10.1080/21645698.2024.2438421. Epub 2024 Dec 24.

ZmNF-YB10, a maize NF-Y transcription factor, positively regulates drought and salt stress response in Arabidopsis thaliana

Yimeng Wang  1 , Peng Jiao  1   2 , Chenyang Wu  1 , Chunlai Wang  1 , Ke Shi  1 , Xiaoqi Gao  1 , Shuyan Guan  1   2 , Yiyong Ma  1   2
Affiliations

ZmNF-YB10, a maize NF-Y transcription factor, positively regulates drought and salt stress response in Arabidopsis thaliana

Yimeng Wang et al. GM Crops Food. 2025 Dec.

Abstract

Maize (Zea mays L.) is a major food and feed crop and an important raw material for energy, chemicals, and livestock. The NF-Y family of transcription factors in maize plays a crucial role in the regulation of plant development and response to environmental stress. In this study, we successfully cloned and characterized the maize NF-Y transcription factor gene ZmNF-YB10. We used bioinformatics, quantitative fluorescence PCR, and other techniques to analyze the basic properties of the gene, its tissue expression specificity, and its role in response to drought, salt, and other stresses. The results indicated that the gene was 1209 base pairs (bp) in length, with a coding sequence (CDS) region of 618 bp, encoding a polypeptide composed of 205 amino acid residues. This polypeptide has a theoretical isoelectric point of 5.85 and features a conserved structural domain unique to the NF-Y family. Quantitative fluorescence PCR results demonstrated that the ZmNF-YB10 gene was differentially upregulated under drought and salt stress treatments but exhibited a negatively regulated expression pattern under alkali and cold stress treatments. Transgenic Arabidopsis thaliana subjected to drought and salt stress in soil showed greener leaves than wild-type A. thaliana. In addition, the overexpression lines showed reduced levels of hydrogen peroxide (H2O2), superoxide (O2-), and malondialdehyde (MDA) and increased activities of peroxidase (POD), catalase (CAT), and superoxide dismutase (SOD). Western blot analysis revealed a distinct band at 21.8 kDa. Salt and drought tolerance analyses conducted in E. coli BL21 indicated a positive regulation. In yeast cells, ZmNF-YB10 exhibited a biological function that enhances salt and drought tolerance. Protein interactions were observed among the ZmNF-YB10, ZmNF-YC2, and ZmNF-YC4 genes. It is hypothesized that the ZmNF-YB10, ZmNF-YC2, and ZmNF-YC4 genes may play a role in the response to abiotic stresses, such as drought and salt tolerance, in maize.

Keywords: Arabidopsis; drought stress; maize; salt stress; yeast heterologous expression.

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Conflict of interest statement

No potential conflict of interest was reported by the author(s).

Figures

Figure 1.
Figure 1.
Bioinformatics analysis of ZmNF-YB10 gene. (a) ZmNF-YB10 interspecies covariance analysis. (b) ZmNF-YB10 intraspecies covariance analysis. (c) ZmNF-YB10 phylogenetic tree analysis.
Figure 2.
Figure 2.
Analysis of ZmNF-YB10 gene expression pattern. (a) Differential expression analysis of ZmNF-YB10 in different maize tissues. (b) Expression pattern analysis of ZmNF-YB10 in roots under alkali treatment (125 mm/L NaHCO3: Na2CO3 = 9:1). (c) Expression pattern analysis of ZmNF-YB10 in roots under drought (300 mm/L mannitol). (d) Expression pattern analysis of ZmNF-YB10 in roots under cold (4°C) treatment. (e) Expression pattern analysis of ZmNF-YB10 in the roots under salt (300 mm/L NaCl) treatment. Student’s t-test was performed; asterisks indicate significant differences, with p < .05 denoted by *and p < .01 denoted by **. Data are expressed as the mean±standard deviation of three independent tests.
Figure 3.
Figure 3.
Efficient expression of ZmNF-YB10 in prokaryotic systems. (a) SDS-page electrophoretic analysis of the ZmNF-YB10 recombinant protein. M, protein marker; N, pEt22b empty vector not induced by IPTG; 0, pEt22b-ZmTCP14 non-induced bacteria; 1–8, pEt22b-ZmNF-YB10 induced for 1–8 h. (b) Protein blot of ZmNF-YB10. M: protein marker; 0: pEt22b-ZmNF-YB10 induced bacteria; 1–4: pEt22b-ZmNF-YB10 induced bacteria.
Figure 4.
Figure 4.
Salt and drought tolerance analyses of ZmNF-YB10 in E. coli BL21.
Figure 5.
Figure 5.
Heterologous expression of ZmNF-YB10 in yeast cells. (a) Tolerance analysis of ZmNF-YB10 in yeast under abiotic stress conditions. (b) Yeast growth curves.
Figure 6.
Figure 6.
Arabidopsis germination rate and phenotypic determinations: (a) germination rate, (b) rosette leaf area, (c) plant height, (d) fruit pod size, (e) root length, (f) length of the sixth leaf blade, (j) leaf changes under stress treatments, (k) leaf length, and (g) leaf phenotypic changes under drought and salt stress treatments. (h) Phenotypic changes in transgenic Arabidopsis plants under drought and salt stress treatments. The Student’s t-test was performed; asterisks indicate significant differences, with p < .05 denoted by * and p < .01 denoted by **. Data are expressed as the mean ± SD of three independent experiments.
Figure 6.
Figure 6.
Continued
Figure 7.
Figure 7.
ROS staining and physiological index detection in ZmNF-YB10 Arabidopsis plants. (a) DAB staining. (b) NBT staining. (c-d) accumulation of H2O2 and O2 - in the leaves of different strains. (e-h) POD, SOD, CAT activities, and MDA assays were determined in Arabidopsis under different stress treatments. The Student’s t-test was performed; asterisks indicate significant differences, with p < .05 denoted by * and p < .01 denoted by **. Data are expressed as mean ± SD of three independent tests.
Figure 8.
Figure 8.
Toxicity validation and self-activation assay of ZmNF-YB10 in yeast cells.
Figure 9.
Figure 9.
Validation of ZmNF-YB10 interaction in yeast.
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