Verticillium wilt is a destructive disease with international consequences for cotton production. Breeding broad-spectrum resistant cultivars is considered to be one of the most effective means for reducing crop losses. A resistant cotton cultivar,60182,was crossed with a susceptible cultivar,Jun-mian 1,to identify markers for Verticillium resistance genes and validate the mode of its inheritance. Genetic segregation analysis for Verticillium wilt resistance was evaluated based upon infected leaf percentage in the seedling stage using major gene-polygene mixed inheritance models and joint analysis of P1,P2,F1,B1,B2 and F2 populations obtained from the cultivar cross. We found that resistance of upland cotton cultivar 60182 to isolates BP2,VD8 and T9,and their isoconcentration mixture was controlled by two major genes with additive-dominance-epistatic effects,and the inheritance of the major gene was dominant. Furthermore,a genetic linkage map was constructed using F2 segregating population and resistance phenotypic data were obtained using F2:3 families inoculated with different isolates and detected in different developmental stages. The genetic linkage map with 139 loci was comprised of 31 linkage groups covering 1165 cM,with an average distance of 8.38 cM between two markers,or 25.89% of the cotton genome length. From 60182,we found 4 QTL on chromosome D7 and 4 QTL on D9 for BP2,5 QTL on D7 and 9 QTL on D9 for VD8,4 QTL on D7 and 5 QTL on D9 for T9 and 3 QTL on D7 and 7 QTL on D7 for mixed pathogens. The QTL mapping results revealed that QTL clusters with high contribution rates were screened simultaneously on chromosomes D9 and D7 by multiple interval mapping (CIM),whether from resistance phenotypic data from different developmental stages or for different isolates. The result is consistent with the genetic model of two major genes in 60182 and suggests broad-spectrum resistance to both defoliating isolates of V. dahliae and nondefoliating isolates. The markers associated with resistance QTL may facilitate the use of Verticillium wilt resistance genes in improving breeding programs for cotton. Verticillium wilt is a destructive disease with international consequences for cotton production. Breeding broad-spectrum resistant cultivars considered to be one of the most effective means for reducing crop losses. A resistant cotton cultivar, 60182, was crossed with a susceptible cultivar, Jun- mian 1, to identify markers for Verticillium resistance genes and validate the mode of its inheritance. Genetic segregation analysis for Verticillium wilt resistance was evaluated based upon infected leaf percentage in the seedling stage using major gene-polygene mixed inheritance models and joint analysis of P1, P2, F1, B1, B2 and F2 populations obtained from the cultivar cross. We found that resistance of upland cotton cultivar 60182 to isolates BP2, VD8 and T9, and their isoconcentration mixture was controlled by two major genes with additive-dominance-epistatic effects , and the inheritance of the major gene was dominant. Furthermore, a genetic linkage map was constructed using F2 segregating popula tion and resistance phenotypic data were obtained using F2: 3 families inoculated with different isolates and detected in different developmental stages. The genetic linkage map with 139 loci was comprised of 31 linkage groups covering 1165 cM, with an average distance of 8.38 cM between two markers , or 25.89% of the cotton genome length. From 60182, we found 4 QTL on chromosome D7 and 4 QTL on D9 for BP2,5 QTL on D7 and 9 QTL on D9 for VD8,4 QTL on D7 and 5 QTL on D9 for T9 and 3 QTL on D7 and 7 QTL on D7 for mixed pathogens. The QTL mapping results revealed that QTL clusters with high contribution rates were screened on chromosomes D9 and D7 by multiple interval mapping (CIM), whether from resistance phenotypic data from different developmental stages or for different isolates. The result is consistent with the genetic model of two major genes in 60182 and suggests broad-spectrum resistance to both defoliating isolates of V. dahliae and nondefoliating isolates. The markersassociated with resistance QTL may facilitate the use of Verticillium wilt resistance genes in improving breeding programs for cotton.