Plant height is a typical quantitative trait and can easily be influenced by genetic background and environmental factors. Single-segment-substitution lines (SSSLs) can effectively reduce the interference of the genetic background. We developed a set of genome-wide SSSLs carrying Nipponbare introgression segments in a background of Guangluai 4. Quantitative trail loci (QTL) analysis involved one-way ANOVA and Dunnett’s test for comparing SSSLs and the recurrent parent Guangluai 4 (P≤0.001). We detected 24 QTLs for plant height distributed on 11 rice chromosomes, except chromosome 10, of which 3 had a negative effect and the others a positive effect. The additive effect of these QTLs ranged from –6.5 to 31.74, with additive effect percentages –8.81% to 41.96%. These results provide useful information for exploring and using new dwarf or semi-dwarf resources.