Conflux + I&E Flux + I&M Flux = 細(xì)胞內(nèi)外離子/分子同時(shí)檢測完整方案 |
活性氧(ROS)是植物體中重要的毒性及調(diào)控物質(zhì),受一系列生物及非生物脅迫、激素、發(fā)育及重力信號(hào)、礦物質(zhì)代謝缺失等誘導(dǎo)產(chǎn)生。目前,人們已經(jīng)發(fā)現(xiàn)了包括MAPK激酶、轉(zhuǎn)錄因子等在內(nèi)的許多活性氧感知系統(tǒng)。此外,質(zhì)膜離子通道也可能參與植物的活性氧感知及活性氧介導(dǎo)的調(diào)控反應(yīng);钚匝醯馁|(zhì)膜離子通道在動(dòng)物關(guān)鍵生理功能中的重要作用已經(jīng)比較明確,但在植物中,此類離子通道特別是羥自由基(HR)激活的K+通道的性質(zhì)及功能尚不清楚。 2010年4月7日,英國的科學(xué)家Demidchik等人選用野生型(WT)及K+外流通道基因缺失突變體(gork1-1)的擬南芥為研究對(duì)象,應(yīng)用非損傷微測技術(shù)、電子順磁共振(EPR)、膜片鉗,以及細(xì)胞成像等方法研究了不同處理?xiàng)l件下擬南芥根及其原生質(zhì)體的K+流速、氧自由基、膜電位、凋亡細(xì)胞形態(tài)等指標(biāo)。研究發(fā)現(xiàn),HR及脅迫均可引起擬南芥幼苗根部產(chǎn)生明顯的K+外流,但突變體中外流明顯較弱。NaCl可誘導(dǎo)根部產(chǎn)生HR進(jìn)而激活K+通道。突變體gork1-1、經(jīng)過K+通道抑制劑或HR清除劑處理過的野生型擬南芥根部細(xì)胞凋亡比較緩慢。由此得出,植物中HR激活的K+通道也參與了細(xì)胞凋亡。 此研究結(jié)果提出了ROS調(diào)控植物陽離子運(yùn)輸?shù)囊粋(gè)新觀點(diǎn),并闡述了HR激活的K+通道在植物中的生理作用。
關(guān)鍵詞:鉀離子(Potassium)、細(xì)胞程序性死亡(Programmed cell death)、 活性氧(Reactive oxygen species)、脅迫(Stress)
參考文獻(xiàn):Demidchik et al. J. Cell Sci. doi: 10.1242/jcs.064352
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ABSTRACT: Reactive oxygen species (ROS are central to plant stress response, signalling, development and a multitude of other processes. In this study, the plasma-membrane hydroxyl radical (HR-activated K+ channel responsible for K+ efflux from root cells during stress accompanied by ROS generation is characterised. The channel showed 16-pS unitary conductance and was sensitive to Ca2+, tetraethylammonium, Ba2+, Cs+ and free-radical scavengers. The channel was not found in the gork1-1 mutant, which lacks a major plasma-membrane outwardly rectifying K+ channel. In intact Arabidopsis roots, both HRs and stress induced a dramatic K+ efflux that was much smaller in gork1-1 plants. Tests with electron paramagnetic resonance spectroscopy showed that NaCl can stimulate HR generation in roots and this might lead to K+-channel activation. In animals, activation of K+-efflux channels by HRs can trigger programmed cell death (PCD. PCD symptoms in Arabidopsis roots developed much more slowly in gork1-1 and wild-type plants treated with K+-channel blockers or HR scavengers. Therefore, similar to animal counterparts, plant HR-activated K+ channels are also involved in PCD. Overall, this study provides new insight into the regulation of plant cation transport by ROS and demonstrates possible physiological properties of plant HR-activated K+ channels. |