This experiment involved the study of effect of 2,4-D on callus induction in PlantagoovataG-2(Isubgol). Blond phyllium (Isubgol)is a medicinal plant that belongs to family Plantaginaceae. Plantago ovatais a winter season herb valued for its mucilaginous husk on its seed for pharmaceutical uses mainly for the constipation and diarrohea.The seeds of Plantago ovatawere procured from IARI New Delhi. The plants were maintained in the Green houses in Bhopal. In vitrosprouts of Plantago ovata G-2 was cultured on MS media supplemented with differ-ent conc. of 2,4-D (0.5mg/l to 5mg/l). Callus induction and photoperiod was maintained 16 hour light and 8 hour darkness and temperaturewas kept 28°C+2°C.Observations were recorded in terms of fresh and dry weight of callus after 40 days of inoculation. The maximum mean fresh weight of callus was obtained from 2.0mg/l of 2,4-D was 1.73gm+-4.1 in Plantago ovataG-2. From the present study it was found that the maximum average fresh weight of callus was seem to be physically friable and yellowish white in color while compact and brownish colored callus were found to be less in amount as compared to healthy callus.