Centro Universitario de Investigaciones Biomédicas, Universidad de Colima, 28045 Colima, Mexico.
Plant Physiol. 2011 Dec;157(4):2167-80. doi: 10.1104/pp.111.179671. Epub 2011 Oct 6.
Reactive oxygen species (ROS) are integral components of the plant adaptive responses to environment. Importantly, ROS affect the intracellular Ca(2+) dynamics by activating a range of nonselective Ca(2+)-permeable channels in plasma membrane (PM). Using patch-clamp and noninvasive microelectrode ion flux measuring techniques, we have characterized ionic currents and net K(+) and Ca(2+) fluxes induced by hydroxyl radicals (OH(•)) in pea (Pisum sativum) roots. OH(•), but not hydrogen peroxide, activated a rapid Ca(2+) efflux and a more slowly developing net Ca(2+) influx concurrent with a net K(+) efflux. In isolated protoplasts, OH(•) evoked a nonselective current, with a time course and a steady-state magnitude similar to those for a K(+) efflux in intact roots. This current displayed a low ionic selectivity and was permeable to Ca(2+). Active OH(•)-induced Ca(2+) efflux in roots was suppressed by the PM Ca(2+) pump inhibitors eosine yellow and erythrosine B. The cation channel blockers gadolinium, nifedipine, and verapamil and the anionic channel blockers 5-nitro-2(3-phenylpropylamino)-benzoate and niflumate inhibited OH(•)-induced ionic currents in root protoplasts and K(+) efflux and Ca(2+) influx in roots. Contrary to expectations, polyamines (PAs) did not inhibit the OH(•)-induced cation fluxes. The net OH(•)-induced Ca(2+) efflux was largely prolonged in the presence of spermine, and all PAs tested (spermine, spermidine, and putrescine) accelerated and augmented the OH(•)-induced net K(+) efflux from roots. The latter effect was also observed in patch-clamp experiments on root protoplasts. We conclude that PAs interact with ROS to alter intracellular Ca(2+) homeostasis by modulating both Ca(2+) influx and efflux transport systems at the root cell PM.
活性氧(ROS)是植物适应环境的重要组成部分。重要的是,ROS 通过激活质膜(PM)中的一系列非选择性 Ca2+通透通道来影响细胞内 Ca2+动力学。我们使用膜片钳和非侵入性微电极离子通量测量技术,表征了豌豆(Pisum sativum)根中羟基自由基(OH•)诱导的离子电流和净 K+和 Ca2+通量。OH•,而不是过氧化氢,激活了快速的 Ca2+外排和更缓慢发展的净 Ca2+内流,同时伴有净 K+外排。在分离的原生质体中,OH•引发了一种非选择性电流,其时间过程和稳态幅度与完整根中的 K+外排相似。这种电流表现出低离子选择性,并且对 Ca2+具有通透性。PM Ca2+泵抑制剂 eosine yellow 和 erythrosine B 抑制了 OH•诱导的根中 Ca2+外排。阳离子通道阻断剂钆、硝苯地平和维拉帕米以及阴离子通道阻断剂 5-硝基-2-(3-苯丙基氨基)-苯甲酸酯和 niflumate 抑制了根原生质体中的 OH•诱导的离子电流以及根中的 K+外排和 Ca2+内流。与预期相反,多胺(PAs)并没有抑制 OH•诱导的阳离子通量。在 spermine 的存在下,净 OH•诱导的 Ca2+外排大大延长,并且测试的所有 PAs(spermine、spermidine 和 putrescine)均加速并增强了 OH•诱导的根净 K+外排。在后一种效应也在根原生质体的膜片钳实验中观察到。我们得出结论,PAs 与 ROS 相互作用,通过调节质膜上的 Ca2+内流和外排转运系统来改变细胞内 Ca2+稳态。