Burdach Zbigniew, Kurtyka Renata, Siemieniuk Agnieszka, Karcz Waldemar
Department of Plant Physiology, Faculty of Biology and Environmental Protection, University of Silesia, Jagiellońska 28, PL-40-032 Katowice, Poland.
Department of Plant Physiology, Faculty of Biology and Environmental Protection, University of Silesia, Jagiellońska 28, PL-40-032 Katowice, Poland
Ann Bot. 2014 Oct;114(5):1023-34. doi: 10.1093/aob/mcu170. Epub 2014 Aug 16.
The mechanism of auxin action on ion transport in growing cells has not been determined in detail. In particular, little is known about the role of chloride in the auxin-induced growth of coleoptile cells. Moreover, the data that do exist in the literature are controversial. This study describes experiments that were carried out with maize (Zea mays) coleoptile segments, this being a classical model system for studies of plant cell elongation growth.
Growth kinetics or growth and pH changes were recorded in maize coleoptiles using two independent measuring systems. The growth rate of the segments was measured simultaneously with medium pH changes. Membrane potential changes in parenchymal cells of the segments were also determined for chosen variants. The question of whether anion transport is involved in auxin-induced growth of maize coleoptile segments was primarily studied using anion channel blockers [anthracene-9-carboxylic acid (A-9-C) and 4,4'-diisothiocyanatostilbene-2,2'-disulphonic acid (DIDS)]. In addition, experiments in which KCl was replaced by KNO3 were also performed.
Both anion channel blockers, added at 0·1 mm, diminished indole-3-acetic acid (IAA)-induced elongation growth by ~30 %. Medium pH changes measured simultaneously with growth indicated that while DIDS stopped IAA-induced proton extrusion, A-9-C diminished it by only 50 %. Addition of A-9-C to medium containing 1 mm KCl did not affect the characteristic kinetics of IAA-induced membrane potential changes, while in the presence of 10 mm KCl the channel blocker stopped IAA-induced membrane hyperpolarization. Replacement of KCl with KNO3 significantly decreased IAA-induced growth and inhibited proton extrusion. In contrast to the KCl concentration, the concentration of KNO3 did not affect the growth-stimulatory effect of IAA. For comparison, the effects of the cation channel blocker tetraethylammonium chloride (TEA-Cl) on IAA-induced growth and proton extrusion were also determined. TEA-Cl, added 1 h before IAA, caused reduction of growth by 49·9 % and inhibition of proton extrusion.
These results suggest that Cl(-) plays a role in the IAA-induced growth of maize coleoptile segments. A possible mechanism for Cl(-) uptake during IAA-induced growth is proposed in which uptake of K(+) and Cl(-) ions in concert with IAA-induced plasma membrane H(+)-ATPase activity changes the membrane potential to a value needed for turgor adjustment during the growth of maize coleoptile cells.
生长素作用于生长中细胞离子转运的机制尚未得到详细确定。特别是,关于氯离子在生长素诱导的胚芽鞘细胞生长中的作用知之甚少。此外,文献中现有的数据存在争议。本研究描述了用玉米(Zea mays)胚芽鞘切段进行的实验,这是研究植物细胞伸长生长的经典模型系统。
使用两个独立的测量系统记录玉米胚芽鞘的生长动力学或生长及pH变化。切段的生长速率与培养基pH变化同时进行测量。还针对选定的变体测定了切段实质细胞中的膜电位变化。主要使用阴离子通道阻滞剂[蒽-9-羧酸(A-9-C)和4,4'-二异硫氰基芪-2,2'-二磺酸(DIDS)]研究阴离子转运是否参与生长素诱导的玉米胚芽鞘切段生长的问题。此外,还进行了用KNO₃替代KCl的实验。
两种阴离子通道阻滞剂在0.1 mM添加时,使吲哚-3-乙酸(IAA)诱导的伸长生长减少约30%。与生长同时测量的培养基pH变化表明,虽然DIDS阻止了IAA诱导的质子外排,但A-9-C仅使其减少50%。在含有1 mM KCl的培养基中添加A-9-C不影响IAA诱导的膜电位变化的特征动力学,而在10 mM KCl存在下,通道阻滞剂阻止了IAA诱导的膜超极化。用KNO₃替代KCl显著降低了IAA诱导的生长并抑制了质子外排。与KCl浓度不同,KNO₃浓度不影响IAA的生长刺激作用。作为比较,还测定了阳离子通道阻滞剂氯化四乙铵(TEA-Cl)对IAA诱导的生长和质子外排的影响。TEA-Cl在IAA前1小时添加,导致生长减少49.9%并抑制质子外排。
这些结果表明Cl⁻在IAA诱导的玉米胚芽鞘切段生长中起作用。提出了IAA诱导生长过程中Cl⁻摄取的一种可能机制,其中K⁺和Cl⁻离子的摄取与IAA诱导的质膜H⁺-ATPase活性协同作用,将膜电位改变为玉米胚芽鞘细胞生长期间膨压调节所需的值。