A major QTL affecting root traits and leaf ABA concentration was identified in maize (Zea mays L.) and named root-ABA1. For this QTL, back-cross-derived lines (BDLs) homozygous either for the (+) or for the (−) allele increasing or decreasing, respectively, root size and leaf ABA concentration, were developed. This study was conducted to evaluate the QTL effects in various genetic backgrounds and at different water regimes. The (+/+) and (−/−) BDLs were crossed with five or 13 inbred tester lines of different origin, thus producing two sets of test-crosses that were evaluated in Italy and China, respectively. Testing was conducted under both well-watered and water-stressed conditions. In Italy, the test-crosses derived from (+/+) BDLs, as compared with those derived from (−/−) BDLs, showed, across both water regimes, higher leaf ABA concentration (on average 384 versus 351 ng g−1 DW) and lower root lodging (28.0 versus 52.5%), and lower grain yield under water-stressed conditions (4.88 versus 6.27 Mg ha−1). In China, where root lodging did not occur, the test-crosses derived from (+/+) BDLs were less productive at both water regimes (on average, 6.83 versus 7.49 Mg ha−1). The lower grain yield of the test-crosses derived from (+/+) BDLs was due to a lower number of ears per plant and to lower kernel weight. The results indicate that the (+) root-ABA1 allele confers not only a consistently lower susceptibility to root lodging but also a lower grain yield, especially when root lodging does not occur.