Crops ›› 2025, Vol. 41 ›› Issue (6): 37-44.doi: 10.16035/j.issn.1001-7283.2025.06.005

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Cloning, Bioinformatics Analysis, and Functional Validation of ZmPOD Gene in Maize

Liu Songtao(), Jiang Chao, Shi Hanbo, Yan Linan, Zhao Haichao, Lu Haibo, Li Hui, Huang Zhihong()   

  1. Hebei North University / The Key Laboratory of Hebei Province Agricultural Products and Food Quality and Safety Analysis and Testing, Zhangjiakou 075000, Hebei, China
  • Received:2024-07-22 Revised:2024-09-29 Online:2025-12-15 Published:2025-12-12

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Abstract:

To investigate the drought resistance mechanisms of maize (Zea mays L.) and to identify and apply drought-resistant genes to improve its drought resistance, this study focused on the maize peroxidase gene ZmPOD, which was previously identified to respond to drought stress. This study cloned the cDNA of ZmPOD gene, performed bioinformatics analysis on its encoded protein?s amino acid sequence, and elucidated the function of the ZmPOD gene by evaluating the phenotypic and physio-biochemical indicators of Arabidopsis overexpression lines under drought stress. The results showed that the ZmPOD gene is 1104 bp in full length, encoding 367 amino acids, and shares the highest homology with peroxidase protein in sorghum. Multiple cis-acting elements were found in the promoter region of this gene, including those related to plant stress response, drought, and abscisic acid (ABA) responsiveness. After drought stress treatment, the root length of overexpressing Arabidopsis lines during germination was significantly higher than that of wild-type (WT) Arabidopsis. During the seedling stage, WT plants exhibited wilting or even death after drought stress, resulting in a lower survival rate compared to the overexpressing lines. Furthermore, after drought treatment, the activities of peroxidase and superoxide dismutase in ZmPOD gene overexpressing lines were higher than in WT, while the malondialdehyde content was lower than in WT. This indicates that overexpression of the ZmPOD gene can enhance plant resistance to drought stress.

Key words: Maize, Drought stress, ZmPOD, Bioinformatics analysis, Functional validation

Fig.1

Results of full-length amplification of ZmPOD gene"

Fig.2

The secondary structure (a) and tertiary structure (b) prediction of ZmPOD protein"

Fig.3

Protein sequence analysis of ZmPOD"

Fig.4

Sequence alignment of ZmPOD proteins in different plants"

Fig.5

PCR result of ZmPOD gene promoter"

Fig.6

Analysis of cis-acting elements of ZmPOD gene promoter"

Fig.7

Drought-resistance identification of ZmPOD overexpression Arabidopsis at germination stage (a) screening of transgenic Arabidopsis positive plants; (c) growth of WT and overexpressing seedlings at different concentrations of mannitol.“*”indicates significant difference from WT (P < 0.05)."

Fig.8

Drought-resistance identification of ZmPOD overexpression Arabidopsis at seedling stage “***”indicates extremely significant difference from WT (P < 0.001)."

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