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李坏死环斑病毒感染对杏种子氧化应激的诱导作用。

Oxidative stress induction by Prunus necrotic ringspot virus infection in apricot seeds.

作者信息

Amari Khalid, Díaz-Vivancos Pedro, Pallás Vincente, Sánchez-Pina María Amelia, Hernández José Antonio

机构信息

Department of Stress Biology and Plant Pathology, CEBAS-CSIC, PO Box 164, 30100 Espinardo, Murcia, Spain.

出版信息

Physiol Plant. 2007 Oct;131(2):302-10. doi: 10.1111/j.1399-3054.2007.00961.x.

Abstract

Prunus necrotic ringspot rvirus (PNRSV) was able to invade the immature apricot seed including the embryo. The amount of virus was very high inside the embryo compared with that present in the cotyledons. PNRSV infection produced an oxidative stress in apricot seeds as indicated by the increase in lipid peroxidation, measured as thiobarbituric acid-reactive substances. This lipid peroxidation increase was parallelled with an imbalance in the seed antioxidant enzymes. A significant decrease in the ascorbate-GSH cycle enzymes as well as in peroxidase (POX) activity took place in infected seeds, suggesting a low capability to eliminate H2O2. No changes in superoxide dismutase (SOD) or catalase activity were observed. A significant decrease in polyphenoloxidase (PPO) activity was also observed. Native PAGE revealed the presence of three different SOD activity bands in apricot seeds: a Mn-containing SOD and two CuZn-containing SODs. Only an isozyme with catalase, glutathione reductase (GR) or PPO activity was detected in both healthy and infected apricot seeds. Regarding POX staining, three bands with POX activity were detected in native gels in both healthy and infected seeds. The gel results emphasise that the drop detected in POX, GR and PPO activities in PNRSV-infected apricot seeds by kinetic analyses was also evident from the results obtained by native PAGE. The oxidative stress and the imbalance in the antioxidant systems from PNRSV-infected apricot seeds resemble the hypersensitive response observed in some virus-host interactions. This defence mechanism would inactivate PNRSV during seed formation and/or the storage period or even during seed germination. Those results can explain the decrease in seed germination and the low transmission of PNRSV by seeds in apricot trees.

摘要

李坏死环斑病毒(PNRSV)能够侵入未成熟的杏种子,包括胚。与子叶中的病毒量相比,胚内的病毒量非常高。如以硫代巴比妥酸反应性物质衡量的脂质过氧化增加所示,PNRSV感染在杏种子中产生了氧化应激。这种脂质过氧化的增加与种子抗氧化酶的失衡同时出现。被感染种子中抗坏血酸 - 谷胱甘肽循环酶以及过氧化物酶(POX)活性显著降低,表明清除H2O2的能力较低。未观察到超氧化物歧化酶(SOD)或过氧化氢酶活性的变化。还观察到多酚氧化酶(PPO)活性显著降低。非变性聚丙烯酰胺凝胶电泳(Native PAGE)显示杏种子中存在三种不同的SOD活性条带:一种含锰SOD和两种含铜锌SOD。在健康和被感染的杏种子中均仅检测到一种具有过氧化氢酶、谷胱甘肽还原酶(GR)或PPO活性的同工酶。关于POX染色,在健康和被感染种子的非变性凝胶中均检测到三条具有POX活性的条带。凝胶结果强调,通过动力学分析在PNRSV感染的杏种子中检测到的POX、GR和PPO活性下降,从非变性聚丙烯酰胺凝胶电泳获得的结果中也很明显。PNRSV感染的杏种子中的氧化应激和抗氧化系统失衡类似于在一些病毒 - 宿主相互作用中观察到的过敏反应。这种防御机制将在种子形成和/或储存期甚至种子萌发期间使PNRSV失活。这些结果可以解释杏树种子发芽率的降低以及PNRSV通过种子的低传播率。

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