Studying bZIP transcription factor 1 gene expression pattern and the activity of some antioxidant enzymes and proline accumulation in maize (Zea mays L.) under salinity stress

Document Type : Original Article

Authors

Department of Plant Production and Genetics, Faculty of Agriculture, Urmia University, Urmia, Iran.

10.22126/cbb.2026.13714.1136

Abstract

Introduction: Maize is a diploid species belonging to the Poaceae family and is classified as a C4 plant in terms of its photosynthetic pathway. Salinity stress results in the increased production of ROS, which in turn triggers a secondary stress known as oxidative stress in plants. Elevated ROS levels specifically induce the peroxidation of unsaturated fatty acids, resulting in the production of secondary products such as ketones, MDA, and other derived compounds. Lipid peroxidation and MDA production, as critical indicators of oxidative damage, are widely utilized to evaluate the degree of sensitivity or tolerance of plant genotypes to salinity stress. To combat salinity-induced oxidative stress, plants activate a suite of enzymatic and non-enzymatic defense mechanisms. Antioxidant enzymes play a fundamental role in ROS detoxification and the maintenance of oxidative homeostasis. Additionally, under salinity stress, the increased synthesis of proline as an active osmolyte facilitates the maintenance of osmotic balance and protects cells against dehydration. The bZIP family represents one of the most extensive groups of transcription factors in plants, playing a crucial role in responding to both biotic and abiotic stresses. This research was conducted to investigate the physiological, biochemical, and molecular responses of maize to salinity stress, focusing on antioxidant enzyme activity, proline accumulation, lipid peroxidation, and the expression pattern of the bZIP transcription factor 1 gene in two tolerant and sensitive lines under a salinity stress.
Materials and methods: Two maize lines, a tolerant line (R10; P14L2) and a sensitive line (S46; MO17), were selected based on the results of previous studies. The experiment was implemented as a factorial based on a CRD with three replications. The first factor consisted of the two maize genotypes, and the second factor included two salinity levels (control and 8 dS/m). Plants were grown in a controlled growth chamber at Urmia University, under temperature conditions of 25°C, a relative humidity of 60–65%, and a photoperiod of 16 hours of light and 8 hours of darkness. Irrigation was performed using Hoagland nutrient solution starting from the two-leaf stage. Salinity stress was applied from the eight-leaf stage for a duration of 20 days using a nutrient solution containing NaCl at a concentration of 8dS/m. Following the stress period, young leaves were sampled for biochemical and molecular analyses, and samples were immediately stored at -80°C. To investigate the expression pattern of the bZIP transcription factor 1 gene, specific primers for the target gene and the reference gene (actin) were designed based on sequences available in the NCBI.
Results: The results demonstrated that salinity stress significantly increased lipid peroxidation, as indicated by malondialdehyde accumulation, as well as proline content in both lines, with a greater magnitude of increase observed in the tolerant line. The activities of superoxide dismutase, ascorbate peroxidase, and catalase were enhanced under salinity stress, whereas guaiacol peroxidase activity showed a decline. Catalase and ascorbate peroxidase exhibited genotype-dependent responses, with higher activity levels recorded in the sensitive line compared to the tolerant line. Correlation analysis revealed significant positive relation among proline content, malondialdehyde levels, and the activities of key antioxidant enzymes. At the molecular level, the expression of the bZIP transcription factor1 gene was significantly affected by time, genotype, and their interaction; the sensitive line displayed an early and transient upregulation of gene expression, while the tolerant line exhibited a delayed but sustained induction of this gene.
Conclusion: Overall, these findings indicate that salinity tolerance in maize is achieved through a coordinated interaction between antioxidant defense regulation, osmolyte accumulation, and temporal control of stress-responsive regulatory gene expression, and that bZIP may serve as a key molecular marker for improving salinity tolerance in maize.

Keywords

Main Subjects

References

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Cereal Biotechnology and Biochemistry
Volume 4, Issue 4 - Serial Number 4
December 2025
Pages 444-472

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