Use of public tropical lines for U.S. commercial maize (Zea mays L.) breeding is either undocumented or non-existent. A possible exception is the old Cuban line A6, which was still being used in tropical hybrids over 40 years after its development. A major reason for the under-utilization of this valuable germplasm source is the sparse amount of yield-trial data available for most tropical lines. Effective evaluation of tropical, unadapted maize is costly and time-consuming in the U.S. corn-belt, where most temperate maize breeding is done. Thus, temperate maize breeding programs have shown minimal interest in such lines. The narrowness of the temperate maize (Zea mays L.) germplasm base has long been recognized, and there are many available, elite tropical lines that might be used to profitably broaden it. However, there are few comparative yield-trial data by which to choose which line(s) might be most useful. As the investment required for using a tropical line in a temperate breeding program is large, line-choice is critical. Tropical maize (Zea mays L.) represents a valuable genetic resource containing unique alleles not present in elite temperate maize. The strong delay in flowering in response to long daylength photoperiods exhibited by most tropical maize hinders its incorporation into temperate maize breeding programs. The objective of this study was to integrate candidate gene analyses with photoperiod QTL mapping across multiple maize populations. We tested the hypothesis that diverse tropical inbreds carry alleles with similar effects at four key photoperiod response quantitative trait loci (QTL) previously identified in maize. Four tropical maize inbreds were each crossed and backcrossed twice to the temperate recurrent parent B73 to establish four sets of introgression lines. Evaluation of these lines under long day lengths demonstrated that all four QTL have significant effects on flowering time or height in these lines, but the functional allelic effects varied substantially across the tropical donor lines. At the most important photoperiod response QTL on chromosome 10, one tropical line allele even promoted earlier flowering relative to the B73 allele. Significant allelic effect differences among tropical founders were also demonstrated directly in an F2 population derived from the cross of Ki14 and CML254. The chromosome 10 photoperiod response QTL position was validated in a set of heterogeneous inbred families evaluated in field tests and in controlled environments.