A high-density linkage map was constructed for an F 2 population derived from an interspecific cross of cultivated allotetraploid species between Gossypium hirsutum L. and G. barbadense L. A total of 186 F 2 individuals from the interspecific cross of "CRI 36 × Hai 7124" were genotyped at 1 252 polymorphic loci including a novel marker system, target region amplification polymorphism (TRAP). The map consists of 1 097 markers, including 697 simple sequence repeats (SSRs), 171 TRAPs, 129 sequence-related amplified polymorphisms, 98 amplified fragment length polymorphisms, and two morphological markers, and spanned 4 536.7 cM with an average genetic distance of 4.1 cM per marker. Using 45 duplicated SSR loci among chromosomes, 11 of the 13 pairs of homologous chromosomes were identified in tetraploid cotton. This map will provide an essential resource for high resolution mapping of quantitative trait loci and molecular breeding in cotton.Key words: cotton; linkage map; target region amplification polymorphism. Available online at www.blackwell-synergy.com/links/toc/jipb, www.jipb.net Genetic linkage maps based on molecular markers have become an important tool for genome analysis, detection of quantitative trait loci (QTL) underlying important traits, physical mapping, map-based cloning and marker-assisted selection. After the first molecular linkage map based on restricted fragment length polymorphism (RFLP) was reported by Reinisch et al. (1994), many maps based on RFLP, RPAD (random amplified polymorphic DNA), SSR (simple sequence repeats) and AFLP (amplified fragment length polymorphism) markers, among others, have been published for segregating populations derived mainly from interspecific (Gossypium hirsutum × Gossypium barbadense) crosses (Shappley et al. 1998; Brubaker et al. 1999; Lacape et al. 2003;Nguyen et al. 2004;Rong et al. 2004;Song et al. 2005;Guo et al. 2006).However, for comprehensive analysis of cotton genome and QTL mapping, it is essential to develop more markers and explore novel molecular marker systems. Expressed sequence tags (EST) have provided an ample source for the development of SSR markers that have been added to the existing cotton linkage maps (Park et al. 2005; Han et al. 2006;Wang et al. 2006). Sequence-related amplified polymorphism (SRAP) marker system, developed by Li and Quiros (2001), has been successfully used to construct cotton linkage maps (Lin et al. 2005;Zhang et al. 2005). Another new marker system, target region amplification polymorphism (TRAP) is also polymerase chain reaction (PCR)-based (Hu and Vick 2003) to detect EST-linked polymorphic markers, in which a fixed primer designed based High-density Linkage Map of Cotton 717 on expressed sequence tags or any cDNAs sequences is used in a PCR reaction in combination with a random primer containing either an AT-or GC-rich core sequence targeting an intron or exon. By using the TRAP technique, Alwala et al. (2006) and Hu et al. (2005) have assessed genetic diversities of sugarcane and lettuce, respectively. L...