A total of 3,200 M5 mutant lines from G. hirsutum breeding line TAM94L25 were characterized for 20 phenotypic traits describing basic plant growth and development, including fiber quality analysis. A subset of 300 lines was selected for further studies. Three replications were grown at single locations in both Texas and Georgia to further evaluate fiber quality, and investigated relationships between trichome variation and fiber traits. Several mutant lines showed substantial improvements over TAM94L25 in economically important traits including fiber elongation, fiber fineness, fiber strength, and fiber length. For each of the traits, multiple lines were identified that consistently had fiber qualities better than the parental line. Further, significant changes in fiber qualities associated with leaf and/or stem trichome mutations suggest that some common genes are involved in trichome and fiber development.