Low temperature is a major abiotic stress for rice cultivation, causing serious yield loss in many countries. To identify QTL controlling low temperature induced spikelet sterility in rice, the progeny of F2, BC1F1 and BC2F1 populations derived from a Reiziq 9 Lijiangheigu cross were exposed to 21/15°C for 15 days at the booting stage, and spikelet sterility was assessed. For genotyping, 92 polymorphic markers from 373 SSR and 325 STS primer pairs were used. A major QTL was initially indentified on the short arm of chromosome 10 by selective genotyping using highly tolerant and susceptible progeny from F2 and BC1F1 populations. The QTL (qLTSPKST10.1) was validated and mapped by genotyping the entire F2 (282 progeny) and BC1F1 (84 progeny) populations.The results from the F2 population showed that qLTSPKST10.1 could explain 20.5% of the variation in spikelet sterility caused by low temperature treatment with additive (a = 14.4) and dominant effect (d = -7.5). From the analysis of 98 selected BC2F1 progeny, the QTL located in the 3.5 cM interval between S10010.9 and S10014.4 was further confirmed. Based on the studies of 3 generations in 2 years, it was clear that the QTL on chromosome 10 is a major determinant of the control of low temperature induced spikelet sterility at booting stage.