中国农业科学 ›› 2021, Vol. 54 ›› Issue (5): 1029-1042.doi: 10.3864/j.issn.0578-1752.2021.05.014

• 园艺 • 上一篇    下一篇

草莓根系线粒体对外源酚酸胁迫的生理响应

鲁晓峰1(),杜国栋2,邵静3,张静茹1,孙海龙1()   

  1. 1中国农业科学院果树研究所/农业农村部园艺作物种质资源利用重点实验室,辽宁兴城 125100
    2沈阳农业大学园艺学院/沈阳市北方果树栽培与生理生态重点实验室,沈阳 110866
    3吉林省农业科学院果树研究所,吉林公主岭 136100
  • 收稿日期:2020-05-21 接受日期:2020-11-26 出版日期:2021-03-01 发布日期:2021-03-09
  • 通讯作者: 孙海龙
  • 作者简介:鲁晓峰,Tel:13314021850;E-mail:13314021850@163.com
  • 基金资助:
    中国农业科学院科技创新工程专项经费(CAAS-ASTIP-201X-ZFRI);中央级公益性科研院所基本科研业务费(1610182020026)

Physiological Response of Mitochondrial Function of Strawberry Roots to Exogenous Phenolic Acid

XiaoFeng LU1(),GuoDong DU2,Jing SHAO3,JingRu ZHANG1,HaiLong SUN1()   

  1. 1Research Institute of Pomology, Chinese Academy of Agricultural Sciences/Key Laboratory of Horticultural Crops Germplasm Resources Utilization, Ministry of Agriculture and Rural Affairs, Xingcheng 125100, Liaoning
    2College of Horticulture, Shenyang Agricultural University/Key Laboratory for Northern Fruit Cultivation and Physiology-Ecology of Shenyang City, Shenyang 110866
    3Pomology Research Institute, Jilin Academy of Agricultural Sciences, Gongzhuling 136100, Jilin
  • Received:2020-05-21 Accepted:2020-11-26 Online:2021-03-01 Published:2021-03-09
  • Contact: HaiLong SUN

摘要:

【目的】连作障碍是设施草莓(Fragaria×ananassa Duch.)生产中遇到的严重问题,根系分泌的酚酸类物质是引起连作障碍的因素之一。本研究旨在探讨连作草莓植株的根系线粒体功能对自毒酚酸物质胁迫的生理响应,为进一步研究草莓连作障碍的生理适应机制奠定基础。【方法】本研究以草莓为试材,分别外源施加27.0 μg·g-1丁香酸、邻苯二甲酸及两者混合溶液,借助电子显微成像、组织化学染色、液相氧电极等技术,观察根系活性氧含量变化,测定线粒体氧化磷酸化水平、呼吸途径及呼吸关键酶活性等指标的变化,研究自毒酚酸物质诱导的活性氧对根系线粒体功能的影响,探究根系线粒体参与的呼吸复合体代谢和能量代谢等生理功能的影响。【结果】施加2种外源酚酸及混合溶液处理均能导致草莓根系活力明显下降,显著影响根系总根长、总表面积、根系总体积和平均直径,抑制根系的发生和生长,抑制作用表现为丁香酸处理(D)>两种溶液混合处理(D+L)>邻苯二甲酸处理(L)>CK。在草莓根系受到外界酚酸胁迫时,随着胁迫时间的延长,H+-ATPase活性前期呈现显著下降趋势,处理第3天时下降幅度最大,处理D、处理L、处理D+L分别比对照下降了38.8%、28.8%、33.7%,阻碍根细胞内外离子的运转能力,后期变化趋于平稳。根系K+Na+-ATPase及Ca2+Mg2+-ATPase活性也一定程度降低,变化规律相似。随着处理时间的延长,根系中超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)及抗坏血酸过氧化物酶(APX)活性均呈现显著下降趋势,H2O2和$ \mathrm{O}_{2}^{\mathop{}_{.}^{-}}$含量逐渐升高,根系线粒体MPT增大,降低Δψm及根系线粒体内膜上细胞色素Cyt c/a含量,破坏线粒体内膜细胞完整性和电子传递链,其中D处理效果最为显著。在草莓植株受到外源酚酸胁迫后,根系呼吸途径虽仍以EMP-TCA途径为主,但贡献率却显著降低,而PPP途径呈现出明显的增强趋势。外源酚酸处理后期,随着胁迫时间的延长,各呼吸关键酶活性均受到抑制作用,呼吸途径比例发生改变,进而导致各呼吸途径受阻,变化程度表现为D>D+L>L>CK。【结论】外源酚酸处理可导致植株根系活力降低,阻碍细胞内外各离子的运转能力,降低根系抗氧化酶活性及根系线粒体功能,各呼吸途径关键酶活性均降低,最终导致各呼吸途径受阻。

关键词: 草莓, 酚酸胁迫, 线粒体, 呼吸, 生理响应

Abstract:

【Objective】Continuous cropping obstacle is a serious problem in the facility production of strawberry (Fragaria × ananassa Duch.). Phenolic acid, a secretion of strawberry roots, is one of the main factors causing continuous cropping obstacle. The aim of this study was to explore the physiological response by which exogenous phenolic acids affected root mitochondrial function in strawberry roots under the effects of phenolic acids accumulated in the rhizosphere of long-term continuous cropping soil. The results were used to discuss the potential responding mechanism of mitochondria in strawberry roots to allelopathy effect mediating by phenolic acid, which would lay the foundation for the further study of continuous cropping obstacle of strawberry.【Method】In this study, strawberries were used as the test material, and syringic acid, phthalic acid and their mixed solution of 27.0 μg·g-1 were exogenously applied. Scanning Electron Microscope (SEM), Histochemical Staining, Liquid-Phase Oxygen Measurement and other techniques were applied to observe the changes of ROS content in root system, to measure the changes of mitochondrial oxidative phosphorylation level, respiratory pathway and the activities of key enzymes in respiration, to study the effect of autotoxic phenolic acid-induced reactive oxygen species on mitochondrial function in root systems, and to explore the effects of root systems’ mitochondria on physiological functions, such as respiratory complex metabolism and energy metabolism. 【Result】Applying two exogenous phenolic acids or mixed solution treatment could significantly reduce the strawberry root vigor, affect the total root length, total surface area, total root volume and average diameter, and inhibit the occurrence and growth of roots, with the degree of changes: syringic acid treatment (D)>two solution mixing treatment (D+L)>phthalic acid treatment (L)>CK. The activity of H+-ATPase decreased significantly in the early stage with the phenolic acid stress time, and the decline amplitude was the largest at the third day. Compared with CK, D, L and D + L were decreased by 38.8%, 28.8% and 33.7%, respectively, which hindered the ion transport capacity of root cells and became stable in the later stage. The activities of K+Na+-ATPase and Ca2+Mg2+-ATPase in roots were also decreased, and the changes were similar. Exogenous phenolic acid could reduce SOD, POD, CAT and APX activity, increase the contents of H2O2 and $ \mathrm{O}_{2}^{\mathop{}_{.}^{-}}$, and also increase MPT in root mitochondria, and reduce Δψm and cytochrome Cyt c/a content in root mitochondrial inner membrane. Although the root respiratory pathway was still mainly the EMP-TCA pathway, the contribution rate was significantly reduced, while the PPP pathway showed an obvious increasing trend. In the late stage of applying exogenous phenolic acids, with the extension of stress time, all respiratory key enzyme activities were inhibited, which led to the obstruction of respiratory pathways, with the degree of changes: D>D+L>L>CK. 【Conclusion】 Exogenous phenolic acid treatment could reduce root vigor and hinder the operational capacity of ions inside and outside cells. The treatment could also reduce root antioxidant enzyme activity and root mitochondrial function, leading to the reduction of key enzyme activities in every respiratory pathways, and ultimately obstruct every respiratory pathways.

Key words: strawberry, phenolic acid stress, mitochondria, respiration, physiological response