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作物杂志,2025, 第6期: 37–44 doi: 10.16035/j.issn.1001-7283.2025.06.005

• 遗传育种·种质资源·生物技术 • 上一篇    下一篇

玉米ZmPOD基因克隆、生物信息学分析及功能验证

刘松涛(), 蒋超, 史涵博, 闫立楠, 赵海超, 卢海博, 栗慧, 黄智鸿()   

  1. 河北北方学院/河北省农产品食品质量安全分析检测重点实验室， 075000， 河北张家口
  • 收稿日期:2024-07-22 修回日期:2024-09-29 出版日期:2025-12-15 发布日期:2025-12-12
  • 通讯作者: 黄智鸿，主要从事作物高产栽培研究，E-mail：hbnuhzh@163.com
  • 作者简介:刘松涛，主要从事作物抗逆基因挖掘与验证研究，E-mail：15028293845@163.com
  • 基金资助:
    河北省教育厅科学技术研究项目(QN2022018)

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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摘要：

为探究玉米（Zea mays L.）的耐旱机制，挖掘并应用耐旱基因以提高其耐旱性，基于前期挖掘到的玉米响应干旱胁迫的过氧化物酶基因ZmPOD，克隆该基因的cDNA，对编码蛋白的氨基酸序列进行生物信息学分析，通过评估干旱胁迫下拟南芥过表达株系的表型和生理生化指标，解析ZmPOD基因的功能。结果表明，ZmPOD基因全长1104 bp，编码367个氨基酸，与高粱中的过氧化物酶蛋白同源性最高。该基因启动子区域存在多个顺式作用元件，包括与植物应激反应、干旱和脱落酸响应相关的元件。干旱胁迫处理后，萌发期过表达拟南芥株系的根长显著高于野生型拟南芥（WT），幼苗期WT经干旱胁迫后出现枯萎表型甚至死亡，存活率低于过表达株系。此外，ZmPOD基因过表达株系的过氧化物酶和超氧化物歧化酶活性均高于WT，丙二醛含量则低于WT，表明过表达ZmPOD基因可增强植物对干旱胁迫的抗性。

关键词: 玉米, 干旱胁迫, ZmPOD, 生物信息学分析, 功能验证

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

图1

ZmPOD基因的全长扩增结果

图2

ZmPOD蛋白的二级结构（a）和三级结构（b）预测

图3

ZmPOD蛋白序列分析

图4

不同植物ZmPOD蛋白序列比对

图5

ZmPOD基因启动子的扩增结果

图6

ZmPOD基因启动子顺式作用元件分析

图7

ZmPOD过表达拟南芥萌发期抗旱性鉴定 （a）转基因拟南芥阳性植株筛选；（c）WT和过表达株系在不同浓度甘露醇中的生长情况。“*”表示与WT差异显著（P < 0.05）。

图8

ZmPOD过表达拟南芥苗期抗旱性鉴定 “***”表示与WT差异极显著（P < 0.001）。

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