The influences of genetic and environmental factors on the anther culture responses of wheat were investigated. Significant differences for callus induction, plant regeneration, and green plant percentages were observed when the nucleus of Triticum aestivum L. cv. Selkirk was transferred to ten alien cytoplasms by substitution backcrosses. In most cases, the alien cytoplasms decreased anther culture responses, but sometimes they were as good as or better than the T. aestivum cytoplasm. Significant within-genotype variation for anther culture responses were observed for wheat varieties Chris, Yecora Rojo, WA7176 and Edwall, indicating genetic heterogeneity in the present commercial cultivars, and potential for improving anther culture responses by in vitro prescreening. When five genotypes (Chris, Pavon 76, Butte 86, WA6916, and Edwall) were cultured across three (potato-4 liquid, 100 g L-1 ficoll-supplemented, and 6 g L-1 agar-solidified) induction media, the liquid and ficoll-containing media were 10 to 15 times more productive than the agar-solidified medium. Whereas, the ficoll medium was not significantly different from the liquid medium. Several low concentration starch media appeared promising to replace current induction media. The starch media sustained the high-callus-induction properties of the liquid medium, while improving callus aeration similar to that observed on solid media, resulting in markedly higher plant regeneration and green plant percentages.