Fiber strength is an essential trait of fiber property in cotton, and it is quantitatively inherited. Identification of stable quantitative trait loci (QTL) contributing to fiber strength would provide the key basis for marker-assisted selection (MAS) in cotton breeding. In this study, four interspecific hybridization populations were established with a common G. barbadense parent Pima 90-53 and two G. hirsutum parents (CCRI 8 and Handan 208), each of which had fiber strength characteristic. Based on the phenotypic data of fiber strength from seven environments, a stable QTL, qFS-c9-1, was detected and validated on c9 in a marker interval between SSR markers NAU2395 and NAU1092. The QTL explaining 14.4-17.9 % of the phenotypic variation was firstly detected in two populations (CCRI 8 × Pima 90-53, BC1F1 and BC1F2) and its derived lines in four environments. And it accounting for 12.1-14.8 % of the phenotypic variation was further confirmed in two populations (Handan 208 × Pima 90-53, BC1F1, and F2) under one environment. In silico mapping using three sequenced cotton genomes indicated that homologous genes, anchored by NAU2395 and NAU1092, were aligned to the G. arboreum genome within a physical distance between 81.10 Mbps and 87.07 Mbps. In that interval, several genes were confirmed in literatures to associate with fiber development. Among these genes, seven genes were further selected for an expression analysis through fiber development transcriptome database, revealing unique expression patterns across different stages of fiber development between CCRI 8 and Pima 90-53. The genes underlying qFS-c9-1 were favorable to fine mapping and cloning. The current study results provided valuable evidence for mapping stable QTL of fiber strength utilizing multiple populations and environments, as well as map-based cloning the candidate gene underlying the QTL for future prospective research directions.