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Plants' roots promote changes in soil structure, forming a strongly-bound soil layer in the surroundings of the root, which is named as rhizosheath. Rhizosheath formation is attributed mainly to the root hairs' presence, that favors the enmeshment of the soil particles around the roots, and the release of mucilage and exudates, which acts as gluing agents of those soil particles. In the present work, we studied the rhizosheath aggregate formation of two Zea mays L. genotypes with contrasting root hair development: a mutant with root hair defective elongation (rth3) and a corresponding wild type (WT). We also tracked the fate of recently-deposited C in the rhizosheath aggregates using two 13CO2 pulse labeling approaches (single vs. multiple pulse labeling). The sampled rhizosheath aggregates were further separated using dry-sieving fractionation into three aggregate size classes: primary small particles and smaller microaggregates (250 µm). We observed that the aggregate size distribution followed the same pattern in both genotypes. This result reinforces the assumption that other soil properties are more important for rhizosheath aggregation than root hair elongation. We observed that the higher potion of the recently-deposited root-derived C (57%) was accumulated in the macroaggregates. Moreover, the multiple pulse labeling approach proportioned a higher 13C enrichment of the rhizosheath aggregates fractions than applying a single pulse. Despite both single and multiple labeling approaches have resulted in a similar distribution of 13C in the rhizosheath aggregates, multiple pulse labeling provided a higher enrichment in the rhizosheath aggregates, which allowed a better separation of significant differences between the genotypes.