Root exudate composition and quantity vary in relation to plant nutritional status, but the impact of the differences on rhizosphere microbial communities is not known. To examine this question, we performed an experiment with barley (Hordeum vulgare) plants under iron-limiting and iron-sufficient growth conditions. Plants were grown in an iron-limiting soil in root box microcosms. One-half of the plants were treated with foliar iron every day to inhibit phytosiderophore production and to alter root exudate composition. After 30 days, the bacterial communities associated with different root zones, including the primary root tips, nonelongating secondary root tips, sites of lateral root emergence, and older roots distal from the tip, were characterized by using 16S ribosomal DNA (rDNA) fingerprints generated by PCR-denaturing gradient gel electrophoresis (DGGE). Our results showed that the microbial communities associated with the different root locations produced many common 16S rDNA bands but that the communities could be distinguished by using correspondence analysis. Approximately 40% of the variation between communities could be attributed to plant iron nutritional status. A sequence analysis of clones generated from a single 16S rDNA band obtained at all of the root locations revealed that there were taxonomically different species in the same band, suggesting that the resolving power of DGGE for characterization of community structure at the species level is limited. Our results suggest that the bacterial communities in the rhizosphere are substantially different in different root zones and that a rhizosphere community may be altered by changes in root exudate composition caused by changes in plant iron nutritional status.Root exudates selectively influence the growth of bacteria and fungi that colonize the rhizosphere by altering the chemistry of soil in the vicinity of the plant roots and by serving as selective growth substrates for soil microorganisms. Microorganisms in turn influence the composition and quantity of various root exudate components through their effects on root cell leakage, cell metabolism, and plant nutrition. Based on differences in root exudation and rhizodeposition in different root zones, rhizosphere microbial communities can vary in structure and species composition in different root locations or in relation to soil type, plant species, nutritional status, age, stress, disease, and other environmental factors (8,16,22,23). During the growth of new roots, exudates secreted in the zone of elongation behind the root tips support the growth of primary root colonizers that utilize easily degradable sugars and organic acids. In the older root zones, carbon is deposited primarily as sloughed cells and consists of more recalcitrant materials, including lignified cellulose and hemicellulose, so that fungi and bacteria in these zones are presumably adapted to crowded, oligotrophic conditions. Other nutritionally distinct sites include the sites of lateral root emergence and the secondary, ...