A previous study on maize F(2:3) families derived from Lo964xLo1016 highlighted one QTL in bin 1.06 (hereafter named root-yield-1.06) affecting root and agronomic traits of plants grown in well-watered (WW) and water-stressed (WS) conditions. Starting from different F(4) families, two pairs of near isogenic lines (NILs) were developed at root-yield-1.06. The objective of this study was to evaluate root-yield-1.06 effects across different water regimes, genetic backgrounds, and inbreeding levels. The NILs per se and their crosses with Lo1016 and Lo964 were tested in 2008 and 2009 near to Bologna, with the well-watered (WW) and water-stressed (WS) treatments providing, on average, 70 mm and 35 mm of water, respectively. For NILs per se, the interactions QTL x water regime and QTL x family were negligible in most cases; the QTL additive effects across families were significant for several traits, especially root clump weight. For NILs crosses, analogously to NILs per se, the interactions were generally negligible and the additive effects across water regimes and families were significant for most traits, especially grain yield. A meta-analysis carried out considering the QTLs described in this and previous studies inferred one single locus as responsible for the effects on roots and agronomic traits. Our results show that root-yield-1.06 has a major constitutive effect on root traits, plant vigour and productivity across water regimes, genetic backgrounds, and inbreeding levels. These features suggest that root-yield-1.06 is a valuable candidate for cloning the sequence underlying its effects and for marker-assisted selection to improve yield stability in maize.