紫花苜蓿老化种子吸胀过程中胚根线粒体抗氧化生理及激素响应规律

doi:10.3864/j.issn.0578-1752.2025.19.016

摘要/Abstract

摘要：

【目的】种子在贮藏过程中不可避免发生老化劣变，主要表现为线粒体功能障碍和活性氧（reactive oxygen species, ROS）累积。线粒体作为细胞能量代谢中心，其抗坏血酸—谷胱甘肽（AsA-GSH）循环是清除ROS的关键防线。探究紫花苜蓿（Medicago sativa L.）胚根线粒体AsA—GSH循环响应老化的抗氧化机制和种子吸胀期间胚根内源激素的变化规律，为揭示紫花苜蓿种子胚根细胞代谢与老化种子萌发过程之间的关系奠定基础。【方法】通过控制劣变处理获得中等活力紫花苜蓿种子，以未老化种子作为对照，研究了种子发芽特性、不同吸胀时间（6、12、24和36 h）胚根线粒体抗氧化酶活性、抗氧化物含量以及不同吸胀时间（12、24和36 h）胚根激素含量的变化规律。【结果】控制劣变处理抑制了紫花苜蓿种子的萌发和幼苗的生长，主要表现为降低种子的发芽势、发芽率和幼苗的根长。在吸胀36 h期间，老化种子胚根线粒体中过氧化氢酶（catalase，CAT）、抗坏血酸过氧化物酶（ascorbate peroxidase，APX）和单脱氢抗坏血酸还原酶（monodehydroascorbate reductase，MDHAR）活性以及抗坏血酸（ascorbic acid，AsA）和谷胱甘肽（glutathione，GSH）含量降低；从吸胀24 h开始，老化种子胚根线粒体中过氧化物酶（peroxidase，POD）和谷胱甘肽还原酶（glutathione reductase，GR）活性升高。随着吸胀的进行，老化和未老化种子AsA/氧化型抗坏血酸（oxidized ascorbic acid, DHA）比值差距渐小，GSH/氧化型谷胱甘肽（oxidized glutathione，GSSG）比值差距渐大。在吸胀过程中胚根脱落酸（abscisic acid，ABA）、赤霉素（gibberellin，GA₃）、生长素（indole acetic acid，IAA）和油菜素内酯（brassinolide，BR）含量的变化趋势基本一致。【结论】老化种子在吸胀过程中，胚根线粒体内的ROS积累，产生氧化损伤，造成种子萌发延迟。老化种子吸胀24 h后，胚根线粒体中POD和GR活性升高，保持一定抗氧化能力和萌发能力。在紫花苜蓿老化种子吸胀萌发过程中，POD和GR活性的增强对于种子活力水平的保持发挥重要作用，为深入探索AsA—GSH循环在抗氧化反应中的作用机制提供了依据，对优化牧草种质资源库的保存策略、延长种子寿命研究具有参考价值。

关键词: 紫花苜蓿, 种子吸胀, 线粒体, 抗氧化系统, AsA—GSH循环, 植物激素

Abstract:

【Objective】Aging deterioration of seeds is inevitable during storage, which is mainly manifested by mitochondrial dysfunction and reactive oxygen species (ROS) accumulation. Mitochondria, as the center of cellular energy metabolism, and its ascorbic acid-glutathione (AsA-GSH) cycle are the key defense to scavenging ROS. This experiment was designed to investigate the antioxidant mechanism of the mitochondrial AsA-GSH cycle and the rules of endogenous hormones changed in the radicle during imbibition of aged alfalfa (Medicago sativa L.) seeds, and to lay a foundation for revealing the relationship between the metabolism of the radicle cells and the imbibed process in the aged alfalfa seeds.【Method】In this experiment, seed samples were treated with controlled deterioration and obtained at a medium level of alfalfa seed vigor, and unaged seeds were used as the control. After the seed germination characteristics were tested, the changes in mitochondrial antioxidant enzyme activity and antioxidant content of the radicle during imbibition (6, 12, 24 and 36 h), and the changes in hormone content of the radicle during imbibition (12, 24 and 36 h).【Result】Controlled deterioration treatment inhibited the seed germination and seedling growth, which mainly presented the decrease in seed germination potential, germination percentage and root length of seedlings. The activities of catalase (CAT), ascorbate peroxidase (APX) and monodehydroascorbate reductase (MDHAR), as well as the contents of ascorbic acid (AsA) and glutathione (GSH) were decreased in the radicle mitochondria of aged seeds within 36 h imbibition; and the activities of peroxidase (POD) and glutathione reductase (GR) increased in the radicle mitochondria of aged seeds after 24 h of imbibition. With the progress of imbibition, the difference of AsA/ oxidized ascorbic acid (DHA) and GSH/ oxidized glutathione (GSSG) ratios between aged and unaged seeds was gradually reduced. The contents of abscisic acid (ABA), gibberellin (GA₃), growth hormone (IAA) and oleuropein lactone (BR) in the radicles presented the similar tendency during the imbibition.【Conclusion】During the process of imbibition, ROS accumulated in the mitochondria of the radicle, resulting in oxidative damage and delayed seed germination. After 24 h imbibition of aged seeds, POD and GR activities in radicle mitochondria increased to maintain a certain antioxidant capacity and germination ability. It could be suggested that the enhancement of POD and GR activities would play an important role in the maintenance of seed vigor level during the imbibition of aged alfalfa seeds, which provided a basis for an in-depth exploration of the mechanism of the role of the AsA-GSH cycle in antioxidant responses, and provided a reference for optimizing the conservation strategy of forage germplasm bank and prolonging seed life.

Key words: alfalfa, seed imbibition, mitochondria, antioxidant system, AsA-GSH cycle, phytohormone

李若鸿, 马馼, 赵士强, 毛培胜. 紫花苜蓿老化种子吸胀过程中胚根线粒体抗氧化生理及激素响应规律[J]. 中国农业科学, 2025, 58(19): 4014-4025.

LI RuoHong, MA Wen, ZHAO ShiQiang, MAO PeiSheng. Antioxidant Physiology and Hormonal Response Pattern of Embryonic Root Mitochondria During Imbibition of Aging Seeds of Alfalfa[J]. Scientia Agricultura Sinica, 2025, 58(19): 4014-4025.

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