<p>This study reported an efficient protocol for <i>in vitro</i> indirect regeneration of clonally true-to-type lentil (<i>Lens culinaris</i> Medik. variety Moitree). Internodal segments were excised and were inoculated in MS (Murashige and Skoog) medium fortified with 12 diverse combinations of different plant growth regulators (PGRs), namely α-naphthaleneacetic acid (NAA), 6-benzyladenine (BA), 2,4-dichlorophenoxyacetic acid (2,4-D), and kinetin (Kin). MS medium in combination with 3.0 mg L⁻¹ 2,4-D and 0.5 mg L⁻¹ Kin (T₁₁) exhibited the fastest callus initiation of 6.5 days (d), while MS medium fortified with 2.0 mg L⁻¹ BA and 1.0 mg L⁻¹ NAA (T₄) produced the highest callusing percentage (98.3%), fresh weight (33.9 mg), and dry weight (3.9 mg). Subsequent shoot regeneration from callus was effective only in MS medium fortified with 2.0 mg L⁻¹ BA and 1.0 mg L⁻¹ NAA (T₄), which resulted in shoot induction and proliferation. Histological analysis of callus cultures using hematoxylin and eosin staining confirmed characteristic developmental stages of friable callus and organogenic callus. For rooting, the regenerated shoots were cultured in simple ½MS medium (without any PGRs). Flow cytometry analysis, along with inter simple sequence repeats and start codon targeted polymorphism molecular markers, exhibited clonal fidelity of <i>in vitro</i> regenerated plantlets. This protocol can be useful for the induction and recovery of the genetically transformed plantlets of lentil in the future.</p>