Identification of responsive genotypes and development of efficient protocols are the prerequisite to an effective doubled haploid production system in applied breeding programs. Evaluation of 16 low linolenic flax (Linola) genotypes/populations with diverse genetic backgrounds from a Linola breeding program using A 22 C medium containing 9% sucrose (A 22 C-9) led to the identification of a number of responsive genotypes. For 96-3-F 1 hybrid, callus induction was greater in modified NLN medium containing 12% sucrose (NLN-12) than in A 22 C-9. But there was no difference in shoot regeneration between NLN-12 and A 22 C-9. For 96-45-F 1 hybrid, there was no difference in callus production between the two media. However, A 22 C-9 had a greater shoot regeneration than NLN-12. In comparison to sucrose, lactose was found to increase callus induction from anthers for all three genotypes tested. However, the effect of lactose on shoot regeneration appeared to be genotype-dependent.Keywords Anther culture · Carbohydrate · Doubled haploid · Linum usitatissimum L.Abbreviations A 22 C-6: A 22 C medium containing 6% sucrose · A 22 C-9: A 22 C medium containing 9% sucrose · BAP: 6-Benzyl-amino-purine · 2,4-D: 2,-4-Dichlorophenoxyacetic acid · NLN-12: Modified NLN medium containing 12% sucrose · PPFD: Photosynthetic photon flux density Communicated by S. Gleddie