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Cotton fiber strength is mainly determined during the secondary cell wall deposition stage when cellulose is synthesized. We obtained cDNA of 20-25 d post anthesis (DPA) fiber from 109 F2 progeny and developed a cotton fiber transcriptome profiling via cDNA-AFLP technology using 37 different primer combinations. The F2 population originated from an interspecific cross between Gossypium hirsutum and Gossypium barbadense. One hundred and thirty-eight absence/presence polymorphic transcript- derived fragments (TDFs), with sizes ranging from 100 bp to 722 bp, were screened. Of these, 75 (53.62%) were polymorphic between the parents of the F2 population. Sequencing the 75 transcripts revealed that 37 of them had been reported to be cotton fiber ESTs. Nine of 75 transcript sequences were homologous to 7 cloned cotton fiber genes, encoding cysteine proteinase, vacuolar H+-pyrophosphatase, vacuolar H+-ATPase, catalytic subunit, arabinogalactan protein, putative receptor protein kinase PERK1, GIA/RGA-like gibberellin response modulator and cellulose synthase. Some other transcripts may represent new gene fragments in cotton fiber development. Surprisingly, 46 of the 75 transcripts were mapped to a single linkage group. The transcriptome groups and the sequenced TDF5 could serve as important resources in the functional genomic research of cotton fiber development. [ABSTRACT FROM AUTHOR]