Kollmeier M, Dietrich P, Bauer C S, Horst W J, Hedrich R
Institute of Plant Nutrition, University of Hannover, Herrenhäuser Strasse 2, D-30419 Hannover, Germany.
Plant Physiol. 2001 May;126(1):397-410. doi: 10.1104/pp.126.1.397.
In search for the cellular and molecular basis for differences in aluminum (Al) resistance between maize (Zea mays) cultivars we applied the patch-clamp technique to protoplasts isolated from the apical root cortex of two maize cultivars differing in Al resistance. Measurements were performed on protoplasts from two apical root zones: The 1- to 2-mm zone (DTZ), described as most Al-sensitive, and the main elongation zone (3-5 mm), the site of Al-induced inhibition of cell elongation. Al stimulated citrate and malate efflux from intact root apices, revealing cultivar differences. In the elongation zone, anion channels were not observed in the absence and presence of Al. Preincubation of intact roots with 90 microM Al for 1 h induced a citrate- and malate-permeable, large conductance anion channel in 80% of the DTZ protoplasts from the resistant cultivar, but only 30% from the sensitive cultivar. When Al was applied to the protoplasts in the whole-cell configuration, anion currents were elicited within 10 min in the resistant cultivar only. La3+ was not able to replace or counteract with Al3+ in the activation of this channel. In the presence of the anion-channel blockers, niflumic acid and 4, 4'-dinitrostilbene-2, 2'disulfonic acid, anion currents as well as exudation rates were strongly inhibited. Application of cycloheximide did not affect the Al response, suggesting that the channel is activated through post-translational modifications. We propose that the Al-activated large anion channel described here contributes to enhanced genotypical Al resistance by facilitating the exudation of organic acid anions from the DTZ of the maize root apex.
为探寻玉米(Zea mays)品种间铝(Al)抗性差异的细胞和分子基础,我们将膜片钳技术应用于从两个铝抗性不同的玉米品种根尖皮层分离的原生质体。在两个根尖区域的原生质体上进行了测量:1至2毫米区域(DTZ),被描述为对铝最敏感,以及主要伸长区(3至5毫米),即铝诱导细胞伸长受抑制的部位。铝刺激完整根尖分泌柠檬酸和苹果酸,显示出品种差异。在伸长区,无论有无铝,均未观察到阴离子通道。完整根系用90微摩尔铝预孵育1小时后,抗性品种80%的DTZ原生质体诱导出一种对柠檬酸和苹果酸通透的、大电导阴离子通道,而敏感品种只有30%。当以全细胞模式将铝施加到原生质体上时,仅在抗性品种中10分钟内就引发了阴离子电流。La3+不能替代或抵消Al3+对该通道的激活作用。在阴离子通道阻滞剂氟尼辛和4,4'-二硝基芪-2,2'-二磺酸存在的情况下,阴离子电流以及分泌速率均受到强烈抑制。应用环己酰亚胺不影响铝反应,表明该通道是通过翻译后修饰被激活的。我们提出,本文所述的铝激活大阴离子通道通过促进玉米根尖DTZ中有机酸阴离子的分泌,有助于增强基因型铝抗性。