Al-induced release of Al-chelating ligands (primarily organic acids) into the rhizosphere from the root apex has been identified as a major Al tolerance mechanism in a number of plant species. In the present study, we conducted physiological investigations to study the spatial and temporal characteristics of Al-activated root organic acid exudation, as well as changes in root organic acid content and Al accumulation, in an Al-tolerant maize (Zea mays) single cross (SLP 181/71 ϫ Cateto Colombia 96/71). These investigations were integrated with biophysical studies using the patch-clamp technique to examine Al-activated anion channel activity in protoplasts isolated from different regions of the maize root. Exposure to Al nearly instantaneously activated a concentration-dependent citrate release, which saturated at rates close to 0.5 nmol citrate h Ϫ1 root Ϫ1 , with the half-maximal rates of citrate release occurring at about 20 m Al 3ϩ activity. Comparison of citrate exudation rates between decapped and capped roots indicated the root cap does not play a major role in perceiving the Al signal or in the exudation process. Spatial analysis indicated that the predominant citrate exudation is not confined to the root apex, but could be found as far as 5 cm beyond the root cap, involving cortex and stelar cells. Patch clamp recordings obtained in whole-cell and outside-out patches confirmed the presence of an Al-inducible plasma membrane anion channel in protoplasts isolated from stelar or cortical tissues. The unitary conductance of this channel was 23 to 55 pS. Our results suggest that this transporter mediates the Al-induced citrate release observed in the intact tissue. In addition to the rapid Al activation of citrate release, a slower, Al-inducible increase in root citrate content was also observed. These findings led us to speculate that in addition to the Al exclusion mechanism based on root citrate exudation, a second internal Al tolerance mechanism may be operating based on Al-inducible changes in organic acid synthesis and compartmentation. We discuss our findings in terms of recent genetic studies of Al tolerance in maize, which suggest that Al tolerance in maize is a complex trait.Al limits crop production on the acid soils that comprise up to 50% of the world's potentially arable lands (von Uexkull and Mutert, 1995). When the soil pH drops below 5, the rhizotoxic Al species, Al 3ϩ , is solubilized into the soil solutions to levels that inhibit root growth and function. However, some plants have evolved Al tolerance mechanisms that enable them to grow in Al-toxic, acid soil environments (for review, see Kochian, 1995;Ma et al., 2001). Earlier work in this field showed that Al tolerance in wheat (Triticum aestivum) was associated with a reduced accumulation of Al in the root apex, but not the mature root (see, for example, Rincon and Gonzales, 1992;Tice et al., 1992; Delhaize et al., 1993a). Delhaize et al. (1993aDelhaize et al. ( , 1993b subsequently provided compelling evidence for the existence of an A...