Lallemantia iberica (Lamiaceae) has pharmaceutical and nutritional importance owing to production of oily seeds and synthesis of commercially secondary metabolites. In the present study, total phenol and flavonoids content and quantitative changes in the shoot and callus pigments of L. iberica were investigated. Moreover, some growth parameters and volatile content of in vitro grown shoots were analyzed. Nodal segments and epicotyldons were used as the explants for shoot and callus formation, respectively. In the multiplication stage of shoot culture, MS medium supplemented with six concentrations of TDZ was employed. For callus induction, 1 mg/l of TDZ, NAA and a combination of NAA and BA were used. Extracted volatile products were analyzed by GC-mass spectrometry. According to the results, the concentration range of TDZ from 0.22 to 0.66 mg/l improved the flavonoids production and the highest content was reported to be in 2 mg/l TDZ (157.5 ± 25.9 mg QE/ g); also, the highest amount of total phenol (3.86 ± 1.02 mg PYRO/g) was observed in 0.66 mg/l TDZ. For induced calli, among the three tested types of PGRs, TDZ treatment was efficient for total phenol and flavonoids. In general, the higher content of flavonoids was obtained from L. iberica shoots while the callus of this plant was efficient for total phenol production. 55 components were identified using GC-MS analysis indicating a low amount of mono and sesquiterpenes and high quantity of alkan hydrocarbons.